Archive for June, 2024

Introduction: iPSCs, an Invaluable Resource for Disease Modeling

The development of human induced pluripotent stem cells (iPSCs) (Takahashi et al., 2007) provided unprecedented opportunities to decipher pathophysiological mechanisms of diseases and to test therapeutic approaches in conditions that better translate to humans. This technology allows to obtain an unlimited number of cells from one patient thus representing an ideal model to study in vitro diseases developmental stages, onset and progression in specific human cells (Park et al., 2008a).

iPSCs are capable of indefinite self-renewal and can differentiate into any cell type under appropriate culture conditions (Takahashi et al., 2007; Yu et al., 2007). iPSCs are generated by reprogramming primary somatic cells, such as dermal fibroblasts or blood cells, using ectopic expression of selected embryonic transcription factors (e.g., Oct4, Sox2, Klf4, and c-Myc) (Takahashi et al., 2007). Over the years, several techniques have been refined to deliver the reprogramming cocktail for iPSCs generation. The first pioneering studies on iPSCs used integrating delivery systems, through retroviral or lentiviral vectors (Takahashi et al., 2007; Yu et al., 2007; Park et al., 2008b). To avoid any incorporation of the foreign genetic material and induction of genomic alterations (Nakagawa et al., 2008; Shao and Wu, 2010), novel delivery systems have been introduced, based on non-integrating vectors (such as the Sendai virus or episomal vectors), self-excising vectors (i.e., Cre-Lox, PiggyBac transposon), and non-viral vectors (i.e., combination of signaling molecules, small bioactive molecules, microRNAs, and other chemicals) (Liu et al., 2020). Interestingly, the delivery of synthetic mRNA expressing the reprogramming factors, was also exploited for the safe generation of iPSCs (Warren et al., 2010). It was also used for iPSCs differentiation (Warren et al., 2012; Mandal and Rossi, 2013; Yoshioka et al., 2013; Goparaju et al., 2017). This technology provides high in vitro transfection efficiency of complex mixtures, with transient expression and absence of genomic integration (Sahin et al., 2014).

iPSCs have the ability to retain the genetic mutation carried by the donor patient together with its genomic background, overcoming the limitations presented by the animal models and leading to a new era of disease modeling and clinical applications (Shi et al., 2017). Moreover, unlike the other unlimited sources of self-renewing cells, the embryonic stem cells (ESCs), which can only be obtained from early-stage blastocysts (45 days post fertilization), the iPSCs can be generated from adult patients, eliminating the ethical issues related to the generation of ESCs and leading to the opportunity for studying different stages of the disorders (Romano, 2008; Romito and Cobellis, 2016).

However, genetic background heterogeneity, lack of proper controls, as well as technical challenges in handling and standardizing the culture methods (Doss and Sachinidis, 2019; Volpato and Webber, 2020), contribute to the variability observed in the use of iPSCs as disease model (Hoekstra et al., 2017; Karagiannis et al., 2018; Volpato and Webber, 2020). To deal with genetic background influence on the expression of disease phenotype it is now possible to generate isogenic cell lines, introducing or repairing putative causative mutations through the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated genomic editing technologies (Ben Jehuda et al., 2018). The use of such controls, when possible, reduces the observed variation in cellular phenotypes caused by the genomic milieu (Soldner and Jaenisch, 2012).

Thanks to the mentioned superior features, iPSCs were exploited to generate in vitro models of severe diseases affecting the neuromuscular system and/or the central nervous system, such as neuromuscular and motor neuron disorders (NMD and MND, respectively). While genetic corrected iPSCs are investigated in the complex field of cell replacement therapies, in which modified cells are reintroduced into patients (Tedesco et al., 2012; Barthlmy and Wein, 2018; Abdul Wahid et al., 2019), the iPSCs platform has already allowed the identification of drug candidates for some of these complex disorders (Ortiz-Vitali and Darabi, 2019; Pasteuning-Vuhman et al., 2020). Recently, the combination of iPSCs and gene targeting approaches is changing the face of modern medicine. In this review, we will thus briefly discuss the successes in the identification of drug candidates for NMD and MND and then we will focus on the efforts toward the validation of gene therapy approaches in iPSCs for muscular dystrophies, amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA). Table 1 summarizes the research efforts in this direction mentioned in this review.

Table 1. Summary of the major findings of the cited articles in which iPSCs were used for therapeutic tests of neuromuscular and motor neuron disorders.

iPSCs are widely exploited in high-throughput drug screenings for genetic disorders. Thus far, the introduction of iPSCs into the drug development pipeline has allowed (i) physiologically improved modeling of disease-relevant phenotypes, (ii) a greater patient stratification, and (iii) discrimination between drug responders and non-responders (Pasteuning-Vuhman et al., 2020). In perspective, this will have an impact on the current limitations of the conventional drug discovery process and consequently improve the success of therapeutic target identification and clinical trial outcomes (Hosoya and Czysz, 2016).

Following their discovery, multiple research efforts focused on the generation of iPSCs for NMD and MND. As example, in 2008 Park and collaborators, established the first iPSCs line from skin fibroblasts from a patient affected by Duchenne muscular dystrophy (DMD), a fatal genetic disorder caused by mutations in the dystrophin (DMD) gene and characterized by progressive muscle wasting (Koenig et al., 1987; Park et al., 2008a; Gao and McNally, 2015). Since then, additional DMD-iPSC lines have been reported by other groups and several differentiation protocols were tested to refine the optimal methods for skeletal muscle and cardiac cell differentiation (reviewed by Danisovic et al., 2018; Piga et al., 2019). These attempts overcame some of the limitations of the commonly used human models of DMD, such as myoblasts obtained from patient biopsies, which are limited in number and phenotypically diverse (Blau et al., 1983; Renault et al., 2000; Sun et al., 2020). In contrast, patient-derived iPSCs allow the generation of large amount of mature skeletal muscle cells (Chal et al., 2016; Caputo et al., 2020) or cardiomyocytesrecapitulating the cardiomyopathy of dystrophic patients (Hashimoto et al., 2016), and can mimic different stages of the disorder (Xia et al., 2018). iPSCs were also converted to neuronal cells to study the impact on the central nervous system in NMD. For example, neuron-iPSCs were generated from patients affected by myotonic dystrophy 1 (DM1) (Du et al., 2013; Xia et al., 2013; Ueki et al., 2017), caused by an expansion of the CTG trinucleotide repeats in the 3 untranslated region of the dystrophia myotonica protein kinase (DMPK) gene (Brook et al., 1992). Altogether these studies highlight the versatility of iPSCs as model for the thorough study of gene mutations in the main affected tissues (i.e., skeletal and cardiac muscle for DMD) but also in other relevant cell types (such as neurons in DM1), which contribute to the disease manifestations. Furthermore, iPSCs are being exploited for the development of therapies for muscular dystrophies which is usually carried out in mouse models unable to fully recapitulate all the human disease features (Wells, 2018; Ortiz-Vitali and Darabi, 2019; van Putten et al., 2020). Recently, Sun and colleagues developed a platform based on DMD-iPSCderived myoblasts for drug screening and among 1524 compounds analyzed, they identified 2 promising small molecules with in vivo efficacy (Sun et al., 2020). Further efforts in this direction will likely improve the search for reliable drug candidates and eventually increase the success rate in clinical trials for these severe disorders.

While animal models remain the preferred choice also for modeling and drug testing for MND (Picher-Martel et al., 2016; Dawson et al., 2018; Giorgio et al., 2019), the large genetic variability of these disorders set the ground for the wide use of patient-derived cells. Since 2008, when Eggans group (Dimos et al., 2008) used for the first time iPSCs to produce patient-specific motor neurons and glia from skin cells of an 82-year-old female patient diagnosed with ALSthe most common adult onset MNDseveral groups have designed and validated protocols for spinal motor neurons (MN) (Son et al., 2011; Amoroso et al., 2013; Demestre et al., 2015; Maury et al., 2015; Toli et al., 2015; Sances et al., 2016; Fujimori et al., 2018) and astrocyte differentiation (Madill et al., 2017; Birger et al., 2019; Zhao et al., 2020). The studies performed in ALS-iPSCs with different genetic mutations, facilitated the identification of common pathological features to the various disease forms, such as endoplasmic reticulum stress (Kiskinis et al., 2014; Dafinca et al., 2016), mitochondrial abnormalities (Dafinca et al., 2020; Hor et al., 2020), and impaired excitability (Wainger et al., 2014), but also characteristics related to specific mutations, like protein aggregation or mislocalization (Liu et al., 2015).

Drug screenings using ALS-derived iPSCs additionally allowed the identification of three drugs that are currently explored as therapeutic options in clinical trials.

- The first one, ROPI, a dopamine receptor agonist, was identified from a panel of 1232 Food and Drug Administration (FDA)-approved drugs in a drug screening analysis conducted at Keio University, which examined Fused in sarcoma (FUS)- and TAR-DNA-Binding Protein 43 (TDP-43)-ALS iPSC-derived MN for suppression of ALS-related phenotypes in vitro, such as mislocalization of FUS/TDP43, stress granule formation, MN death/damage, and neurite retraction (Fujimori et al., 2018). This drug is now tested in the ROPALS trial (UMIN000034954 and JMA-IIA00397) as continuation of the Phase I/IIa clinical trial (Morimoto et al., 2019).

- Retigabine (known as an antiepileptic) was identified as a potential suppressor of the hyperexcitability of ALS iPSC-derived MNs based on electrophysiological analysis (Wainger et al., 2014). It is a voltage-gated potassium channel activator (Kv7) able to both block hyperexcitability and improve MN survival in vitro when tested in ALS cases carrying the most common genetic mutations (Wainger et al., 2014). A Phase II Pharmacodynamic Trial of Ezogabine (Retigabine) on neuronal excitability in ALS (NCT02450552) was conducted from 2015 to 2019 showing a decrease of cortical and spinal MN excitability in participants with ALS. These data suggest that such neurophysiological metrics may be used as pharmacodynamic biomarkers in multisite clinical trials (Wainger et al., 2020).

- The third drug is Bosutinib, a proto-oncogene non-receptor protein tyrosine kinase (Src/c-Abl) inhibitor that promoted autophagy and rescued degeneration in iPSC-derived MN, inhibiting misfolded Superoxide Dismutase 1 (SOD1) aggregation and suppressing cell death in genetic and sporadic ALS (Imamura et al., 2017). A new Phase I clinical trial of the drug bosutinib for ALS (UMIN000036295) was initiated in Japan in March 2019.

These examples of drug discovery in iPSCs and their ongoing translation to patients affected by a yet uncurable disease, indicate that this could be a valid paradigm for clinical success in similar diseases, such as SMA. SMA is a MND caused by homozygous mutations in the survival of motor neuron gene (SMN1) leading to infant mortality and motor disabilities in young and adult patients (Lefebvre et al., 1995; Verhaart et al., 2017; Smeriglio et al., 2020). This gene has a paralog called SMN2 that is nearly identical to SMN1, with few nucleotide differences, which result in the exclusion of exon 7 and 90% production of a truncated non-functional survival of motor neuron (SMN) protein (Lefebvre et al., 1995). Several therapeutic strategies have been tested to restore SMN expression (Wirth, 2021). Histone deacetylase (HDAC) inhibitors were tested to induce transcriptional activation of SMN2 and consequent increased production of full length SMN, with successful outcomes in proof-of-concept studies and failure in clinical trials. With the aim to identify compounds with higher efficacy and specificity, Lai and colleagues performed a drug screening in neuron-iPSCs from SMA patients. This study identified novel HDAC inhibitors with therapeutic potential that could be further explored for SMA treatment (Lai et al., 2017). Interestingly, neuron-iPSC from SMA patients were also used to test the efficacy of the recent FDA approved small molecule EvrysdiTM (risdiplam) (Ratni et al., 2016; Ratni et al., 2018; Dhillon, 2020), which forces the inclusion of exon 7 and thus restore SMN protein levels (Poirier et al., 2018). Moreover, the drug called TEC-1 (2-(4,6-dimethylpyrazolo[1,5-a]pyrazin-2-yl)-6-(4-methylpiperazin-1-yl)quinazolin-4(3H)-one) another SMN2 splicing modulator, was recently identified in a screening on SMA patient-derived fibroblasts. The drugs effects were then confirmed in SMA-MN-iPSCs (Ando et al., 2020).

As suggested by the reported examples, the combination of iPSCs modeling, together with high-throughput drug screening followed by animal tests will likely ensure the identification of effective and safe therapeutic candidates. How this pipeline can be adapted to the development and tests for precision medicine approaches, such as gene therapy, will be discussed in the following paragraphs and is exemplified in Figure 1.

Figure 1. Test and development of gene targeting approaches using iPSCs. This drawing summarizes the steps of development for drugs and gene therapy approaches, using induced pluripotent stem cells (iPSCs). Somatic cells, such as fibroblasts or blood cells (peripheral blood mononuclear cells, PBMCs) are obtained from patients biopsies. After reprogramming, the patient-derived iPSCs can be differentiated into disease-relevant cell types, such as skeletal muscle cells, neural or glial cells for neuromuscular or motor neuron disorders. These cells are then subjected to the classical high-throughput drug screening and in perspective will be used to test novel therapeutic entities, based on gene targeting approaches. As example, antisense oligonucleotides (ASOs) or adeno-associated viral vectors (AAV)-based strategies. After validations in animal models and the pre-clinical development process, these novel therapies could enter into clinical trials for patients affected by rare disorders. The use of iPSCs and gene targeting strategies will likely foster the development of personalized medicine approaches. Created with BioRender.com.

Gene targeting approaches are based on the direct correction of the genetic defects (Wang and Gao, 2014; Cappella et al., 2019). For example, antisense oligonucleotides (ASOs) widely tested in pre-clinical and clinical settings, have been approved for SMA (Spinraza) (Aartsma-Rus, 2017) and DMD (i.e., Exondys 51) (Stein, 2016) patients, encouraging their use for the treatment of other monogenic disorders.

ASOs are synthetic single-stranded strings of nucleic acids that bind to RNA through standard WatsonCrick base pairing. After binding to the targeted RNA, the antisense drug can modulate the function of the targeted RNA by several mechanisms (Bennett and Swayze, 2010; Crooke et al., 2018), depending on the chemical modifications and the binding position on the target RNA (Wurster and Ludolph, 2018; Talbot and Wood, 2019; Ochoa and Milam, 2020). Briefly, ASOs can promote degradation of the targeted RNA, by mimicking DNA-RNA pairing and activating endogenous nucleases (i.e., RNase H1), or can modulate the processing of the RNA molecule, without inducing its degradation. This can be achieved through several mechanisms, such as by masking RNA splicing sites, as in the examples described below for DMD or SMA (Dick et al., 2013; Shoji et al., 2015; Osman et al., 2016; Ramirez et al., 2018). Other methods of action of ASOs have been previously reviewed (Bennett and Swayze, 2010; Crooke, 2017).

Several strategies, (Miller and Harris, 2016; Schoch and Miller, 2017), are currently investigated to increase ASOs stability, enhance binding affinity to the target RNA, improve tissue distribution and cellular uptake, while decreasing possible adverse effects (Bennett et al., 2017). Here we will focus on the use of iPSCs as model for testing the efficacy of these gene targeting approaches in NMD and MND.

Due to the large size of the DMD gene (Koenig et al., 1987), the restoration of the full-length dystrophin protein is challenging (Gao and McNally, 2015; Duan, 2018). One of the most promising approaches for gene targeting in DMD, is the use of ASOs binding to the pre-mRNA of the DMD gene to restore its reading frame and consequently producing a truncated but yet functional protein.

The ASO-mediated exon-skipping efficacy on exon 51 was tested in cardiomyocytes derived from iPSCs with DMD mutations, restoring dystrophin to nearly 30% of the normal level (Dick et al., 2013). Another similar study tested an ASO forcing exon 45 skipping of the DMD gene in myotubes derived from iPSCs, thus restoring dystrophin expression but also reducing calcium overflow (Shoji et al., 2015). These studies indicate that iPSCs can be used as platforms for therapeutic selection of ASO, based on the gene correction and prevention of skeletal muscle phenotype in DMD. The new frontier for the treatment of DMD patients is the development of mutation-specific ASOs (Schneider and Aartsma-Rus, 2020) and the use of iPSCs will likely speed the path to success of those strategies through the selection of the patient-specific and most efficient candidates.

ASOs were also proven effective in differentiated myotubes from DM1-iPSCs. A repeat-directed ASO treatment abolished RNA foci accumulation and rescued mis-splicing (Mondragon-Gonzalez and Perlingeiro, 2018) in vitro. These discoveries indicate that once established the proper conversion and differentiation protocols, together with valid disease read-outs, the test of ASOs in iPSCs could be likely applied to a larger spectrum of muscular dystrophies and diseases.

Therapeutic ASOs are currently tested in clinical trials for ALS patients harboring the chromosome 9 open reading frame 72 (C9ORF72) mutations (NCT03626012), SOD1 mutations (NCT03070119, NCT02623699) (recently reviewed by Cappella et al., 2021) or for sporadic ALS patients, with the Ataxin2-ASO (NCT04494256, Becker et al., 2017). Importantly, a splice switching ASO targeted to SMN2 (Spinraza) was approved for SMA patients in 2016.

To better characterize ASOs ability to rescue disease hallmarks, to dissect pathophysiological mechanisms and to test novel chemistries and molecular technologies, different research groups are studying ASOs in iPSCs for MND. For example, ASOs were proven effective in reducing the accumulation of sense RNA foci or toxic dipeptides in C9ORF72-iPSCs differentiated to neurons or MN (Donnelly et al., 2013; Sareen et al., 2013; Giorgio et al., 2019). More recently, Zhang et al. (2018) demonstrated that nucleocytoplasmic transport deficits and neurodegeneration were alleviated in C9ORF72-MN-iPSCs, after treatment with ASOs directed against the Ataxin 2, an RNA-binding protein. Nizzardo et al. (2016) treated ALS MN-iPSCs with ASOs designed to reduce the synthesis of human SOD1 and observed an increased survival and reduced expression of apoptotic markers in treated cells.

In SMA, iPSCs were used to test novel ASO sequences for their improved capacity of producing the full length SMN protein from splicing modulation of SMN2 and exon 7 inclusion (Osman et al., 2016; Ramirez et al., 2018). They were also used to test novel molecular strategies to restore SMN expression and correct neuropathological feature, namely an U1 small nuclear RNA-mediated splice switching approach and SMN transcription activation, via the Transcription Activator-Like Effector-Transcription Factor (TALE-TF) (Nizzardo et al., 2015). This report suggests that iPSCs could serve for the side-by-side comparison of different gene targeting strategies for monogenic disorders.

The use of adeno-associated viral vectors (AAV) for gene therapy of rare disorders recently became a clinical reality. The approval of Zolgensma (an AAV-mediated therapy) for the treatment of the most severe form of SMA, endorses the development of similar approaches for NMD and MND. Indeed, several pre-clinical studies report successes of these approaches in disease models (Biferi et al., 2017; Cappella et al., 2019; Crudele and Chamberlain, 2019) and their use in clinical trials (Bowles et al., 2012; Mendell et al., 2015; Mueller et al., 2020).

Some of the challenges associated to the translation of AAV-based therapies from animal models to patients, are linked to (i) the selection of the best AAV serotype for efficient transgene expression, (ii) cell/tissue specificity, as well as (iii) production of high vector titers, and (iv) reduction of immunoreactivity (Colella et al., 2017; Naso et al., 2017). To date, hundreds of natural AAV serotypes, variants and bio-engineered versions have been described (Hester et al., 2009; Choudhury et al., 2016; Deverman et al., 2016; Chan et al., 2017; Hanlon et al., 2019). Beside serotypes, research efforts are also focusing on the combination of the best serotype with the therapeutic and regulatory sequencessuch as promoters or enhancers (Colella et al., 2018; Besse et al., 2020; Nieuwenhuis et al., 2020), for efficient, safe and specific transgene expressions (Guilbaud et al., 2019; Hanlon et al., 2019). This will likely contribute to expedite the translational path from bench to clinic. In this context, iPSCs can be used to select the vector with best transduction properties for a specific cell type and/or to test the therapeutic sequences (recombinant transgene, oligonucleotides, antibodies, etc.). These techniques will be further refined to design patient-specific approaches. In perspective, when a therapeutic candidate will be established, iPSCs could be further used for analytical tests of approved gene therapies, such as potency assays.

AAV vectors were initially tested for genetic manipulation of ESCs or iPSCs in vitro, using natural human-derived AAV serotypes (from 1 to 9). After some unsuccessful attempts (Smith-Arica et al., 2003; Jang et al., 2011), some reports showed that natural AAV vector serotypes, such as AAV 2 and 3, were able to target iPSCs, although with limited efficacy (Mitsui et al., 2009; Khan et al., 2010). Through direct evolution, Asuri et al. (2012), derived a novel variant of AAV (AAV1.9) with a threefold higher gene delivery efficiency than AAV2 in iPSCs. These pioneer studies suggested that AAV vectors could be also used for stem cell correction and consequently studies of biological mechanisms in vitro and eventually for therapeutic purposes in cell therapy approaches.

Several studies reported method for AAV-mediated delivery of differentiated iPSCs. For example, Rapti et al. (2015) compared the transduction efficiency of different AAV (serotypes 1, 2, 6, and 9) in cardiomyocyte-iPSCs. Interestingly, they noticed that AAV vectors preferentially transduced differentiated cells and identified in serotypes 2 and 6 the best suited for cardiomyocyte-iPSCs transduction.

For modeling and therapeutic testing of central nervous system cells, AAV serotype 5 expressing the green fluorescent protein (GFP), was proven efficient in iPSCs-derived neuronal and glial cells, resulting in up to 90% of transduction (Martier et al., 2019a). Moreover, Duong et al. examined the level of AAV-GFP expression following the transduction of 11 AAV vectors in iPSCs differentiated into retinal pigment epithelium and cortical neurons (Duong et al., 2019). GFP-expressing cells were examined and compared across doses, time and cell type. They reported that retinal pigmented epithelium had the highest AAV-mediated GFP expression compared to cortical neurons-iPSCs and that AAV7m8 and AAV6 were the best performing, across vector concentrations and cell types. This study suggested that in addition to vector tropisms, cell type significantly affects transgene expression (Duong et al., 2019).

Overall, following optimizations, AAV vectors can be used to efficiently transduce patient-derived cells converted to neural or glial cells, likely facilitating studies for neurological diseases. Indeed, Martier and colleagues investigated the feasibility of a miRNA-based gene therapy to obtain long-term silencing of the repeat-containing transcripts of C9ORF72. Four AAV5 carrying miR candidates were tested in neuron-iPSC, resulting in sufficient transduction and expression of therapeutically relevant levels of the corresponding mature miRNA (Martier et al., 2019b). Two of the tested candidates were then proven efficient in reducing RNA foci accumulation in some brain regions of a disease mouse model (Martier et al., 2019a).

Novel methods are currently developed to select AAV for their fitness in vitro. For example, the group of Lisowski developed an AAV Testing Kit, as novel high-throughput approach based on next-generation sequencing, to study the performance of 30 published AAV variants in vitro, in vivo, and ex vivo. They tested AAV variants in primary cells, immortalized cell lines and iPSCs, showing that iPSCs were most efficiently transduced with bioengineered vectors, such as AAV 7m8, AAV LK03, and AAV DJ (Westhaus et al., 2020). This suggests that further methods for AAV optimization are necessary and will likely improve AAV transduction properties in vitro and in vivo.

Transduction properties of AAV serotypes in the human context have been recently tested in 3D structure iPSC-derived cerebral organoids. The transduction properties of two commonly used AAV serotypes (AAV5 and 9) were compared for transgene expression at the mRNA and protein levels, together with the presence of viral DNA. This study reported a higher transduction of the AAV5 compared to AAV9, in organoids and neural cells (Depla et al., 2020). This work set the ground for the use of iPSCs-derived human organoids as valid system for testing AAV properties and will be likely a valuable platform for holistic characterization of AAV properties in vitro and identification of the best therapeutic candidates.

Gene therapy treatments are revolutionizing the face of modern medicine opening treatment perspectives for patients affected by fatal conditions. Despite the growing success of these approaches, several aspects of gene therapy development need refinement and would benefit of the use of iPSCs. Indeed, together with their most known use, such as disease modeling for high-throughput drug screenings, they can be converted into a reliable platform for testing the novel therapeutic entities. Indeed, after the establishment of proper differentiation protocols and disease readouts, patient-derived models are being utilized to test gene targeting approaches. Here, we have summarized research efforts in testing drugs and gene therapy approaches in iPSCs from patient affected by neuromuscular and motor neuron diseases. We have presented some of the successes in candidate drug identification, such as risdiplam for the treatment of SMA and the research efforts in testing ASOs and AAV-mediated therapies. These studies set the ground for further developments, to select optimized therapeutic molecules and to identify powerful and safe AAV vectors.

In parallel to iPSCs development, research efforts are currently focused on the generation of even more advanced disease models. Indeed, despite iPSCs represent a reliable model for the understanding of pathological mechanisms and therapeutic development, they do not fully recapitulate the complexity of a tissue, with its architecture and interactions (Costamagna et al., 2019). In this direction, 3D culture methods are being implemented for NMD and MND, for example with the generation of artificial skeletal muscle for DMD (Maffioletti et al., 2018) or spinal cord organoids for SMA, which were used for drug test (Hor et al., 2018). Interestingly, the group of Pasa, has recently reported the generation of iPSC-derived 3D culture, in which cerebral cortex or hindbrain/spinal cord organoids were assembled with skeletal muscle spheroids (Andersen et al., 2020). These so-called 3D cortico-motor assembloids hold promise for the development of effective therapeutics for NMD and MND.

In conclusion, the advances in novel technologies, such as production of mature organoids, will endorse the development of efficient personalized medicine approaches.

MC and SE: writing of the manuscript draft. MB: conceptualization, writing, and review. All authors contributed to the article and approved the submitted version.

MC was supported by the ANR grant no. ANR-19-CE18-0014-01. MB and SE were supported by the Association Institut de Myologie (AIM)

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

We thank the Association Franaise contre les Myopathies (AFM), the Association Institut de Myologie (AIM), the Sorbonne Universit, the Institut National de la Sant et de la Recherche Mdicale (INSERM). We also thank Piera Smeriglio for critical reading of the manuscript.

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The Potential of Induced Pluripotent Stem Cells to Test Gene Therapy Approaches for Neuromuscular and Motor ... - Frontiers

Creative Medical Technology Holdings (NASDAQ: CELZ) has successfully generated human insulin-producing Islet Cells derived from induced pluripotent stem cells (iPSC) under its iPSCelz program. This development is validated by Greenstone Biosciences and utilized in several FDA-cleared clinical programs in the U.S. The creation of these cells marks a significant milestone for the company, potentially accelerating clinical applications and saving years of research and development. CEO Timothy Warbington highlighted the cost-efficiency and regulatory adherence of the company's multiple programs while maintaining a lower burn rate compared to peers.

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PHOENIX, June 24, 2024 (GLOBE NEWSWIRE) -- Creative Medical Technology Holdings, Inc. (Creative Medical Technology or the Company) (NASDAQ: CELZ), a leading commercial stage biotechnology company focused on a regenerative approach to immunotherapy, urology, neurology, and orthopedics, today announced that it has successfully generated human induced pluripotent stem cells (iPSC)-derived Islet Cells that produce human insulin.

The iPSC clinical line that generated these insulin producing Islet Cells is part of the Companys iPSCelz program, which is validated by Greenstone Biosciences Inc. (Greenstone). The iPSC clinical line, which is currently utilized in a number of our FDA cleared clinical programs in the U.S., has also been utilized to derive validated mesenchymal cells and T-regulatory cells.

Timothy Warbington, President and CEO of the Company, commented, The production of human insulin from islets derived from the IPSCelz program is a significant milestone for the Creative Medical Team and a reflection of the leadership role we have assumed in developing these therapies. It was only year ago that we confirmed the development of our iPSC. As we said then, we estimated that the development of this cell line would save the Company two to three years in research and development time along with associated expenses. Today, we are thrilled to be able to announce the evolution of this program with the creation of insulin producing Islet Cells derived from our iPSC. We believe that this development has the potential for not only clinical translation of the human Islet Cells, but also the stand-alone human insulin which is produced by these cells. We are currently in strategic discussions on next step collaborations to further these programs.

The Company continues to achieve significant milestones with its multiple programs in a cost-efficient manner without sacrificing quality and maintaining strict adherence to all regulatory requirements, Mr. Warbington continued. We are focused on allocating our resources in a prudent and effective manner which we believe is evidenced by our achievements and a slower burn rate than many companies in our space.

About IPSCelz iPSCelz, which is protected by trade secrets and published U.S. patents, utilizes the companies xeno-free human perinatal cell line derived from qualified human donors which are then converted into IPS cells.These cells are incubated with the Companys cell-free reprogramming cocktail to create the human islets and other cell types.

About Creative Medical Technology Holdings Creative Medical Technology Holdings, Inc. is a commercial stage biotechnology company specializing in stem cell technology in the fields of immunotherapy, urology, neurology, and orthopedics. For further information about the Company, please visit http://www.creativemedicaltechnology.com.

Forward Looking Statements This news release may contain forward-looking statements including but not limited to comments regarding the timing and content of upcoming clinical trials and laboratory results, marketing efforts, funding, etc. Forward-looking statements address future events and conditions and, therefore, involve inherent risks and uncertainties. Actual results may differ materially from those currently anticipated in such statements. See the periodic and other reports filed by Creative Medical Technology Holdings, Inc. with the Securities and Exchange Commission and available on the Commission's website at http://www.sec.gov.

Creative Medical Technology announced the successful generation of human insulin-producing Islet Cells derived from iPSCs under its iPSCelz program.

Creative Medical Technology announced this development on June 24, 2024.

The generation of insulin-producing Islet Cells marks a key milestone, potentially speeding up clinical applications and saving years of research and development time for CELZ.

The iPSCelz program is validated by Greenstone Biosciences.

The PR highlights cost-efficient program management, regulatory adherence, and a slower burn rate compared to peers, as financial benefits for CELZ.

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Creative Medical Technology Holdings Announces Evolutionary Development of its iPSCelz Program with the ... - StockTitan

Mehdi Nikkhah, an associate professor of biomedical engineering in theIra A. Fulton Schools of Engineeringat Arizona State University, and his collaborators at Mayo Clinic in Arizona have been awarded a $2.7 million grant by the National Institutes of Health to research how stem cell engineering and tissue regeneration can aid in heart attack recovery.

The research will be conducted in collaboration withWuqiang Zhu, a cardiovascular researcher and professor of biomedical engineering atMayo Clinic.

Nikkhah and Zhu are exploring stem cell transplantation to repair and possibly regenerate damaged myocardium, or heart tissue. Their work is focused on the development of a new class of engineered heart tissues with the use of human-induced pluripotent stem cells, or hiPSCs, and has resulted in two published papers in ACS Biomaterials.

Aheart attack, medically termed as a myocardial infarction, occurs when a coronary artery that sends blood and oxygen to the heart becomes obstructed. This blockage is often the result of an accumulation of fatty cholesterol-containing deposits, known as plaques, within the hearts arteries.

When these plaques rupture, a cascade of events is initiated, leading to the formation of a blood clot. These blood clots can obstruct the artery, impeding blood flow to the heart muscle, thus triggering a heart attack.

When someone has a heart attack, a portion of muscle tissue on the left ventricle, which pumps the blood throughout the whole body, is damaged, Nikkhah says. Over time, the other parts of the heart have to take on more workload, consequently leading to catastrophic heart failure.

A team of biomedical engineers in theSchool of Biological and Health Systems Engineering, part of the Fulton Schools, and medical researchers at Mayo Clinic in Arizona are taking a novel step forward in using stem cell technology and regenerative medicine to aid in heart attack recovery.

Nikkhah is developing engineered heart tissues, or EHTs, with electrical properties to simulate the contraction function typically found within the native hearts tissue.

He is integrating the EHTs with gold nanorods to enhance electrical conductivity among stem cells. Gold is a suitable material because it is conductive and nontoxic to human cells, making the nanorods safe for medical research and translational studies.

In the lab, Nikkhahs team mixes the gold nanorods with a biocompatible hydrogel to form a tissue construct a patch of stem cells to rejuvenate damaged cardiac muscle tissue, offering a promising outcome for heart regeneration.

After we generate the patch, we get the engineered hiPSCs from Dr. Zhus lab at Mayo Clinic, Nikkhah says. They seed the cells on the patch and look at their biological characterization, including cell proliferation, cell viability and gene expression analysis, to see how the cells respond to the conductive hydrogel.

We have successfully used hiPSC-derived cardiomyocytes and cardiac fibroblasts to create beating heart tissues.

The successful integration and proliferation of these cells can lead to the formation of new, healthy heart tissue, potentially reversing the damage caused by the heart attack and enhancing the recovery process.

Reprogrammed human stem cells have nearly limitless potential because they can be differentiated into various cell types. That means hiPSCs can also be used to construct capillaries and blood vessels, which are essential for restoring adequate blood flow and oxygen supply to the damaged areas of the heart.

This process involves the differentiation of hiPSCs intoendothelial cells, which form the lining of blood vessels, thereby facilitating the reconstruction of the hearts vascular network.

Michelle Jang, a graduate student in Nikkhahs lab, is currently studying EHTs to improve cell maturation and observe its electrical properties.

My engagement in this project showed a deep interest in how biomedical engineering technology and biology intersect to create new therapeutic possibilities in the field of regenerative medicine, Jang says. Im excited to see how my current research will further evolve and potentially contribute valuable insights to biomedical research.

Using these techniques, Nikkhah and Zhu can observe the capacity of programmed cells to regenerate damaged heart tissue. With continued advancement in regenerative medicine, there is potential for significant positive impact on outcomes for patients suffering from heart attacks.

Originally posted here:
Harnessing benefits of stem cells for heart regeneration | ASU News - ASU News Now

Animal models have shown that cardiomyocytes can indeed regenerate. However, animal studies are not always directly transferable to a human setting. Thus, it is essential to assess regenerative processes in human heart when possible. In this study, we evaluated the expression of several early cardiac stem cell- and proliferation- associated biomarkers in adult human cardiac tissue from the left ventricle (LV) and the potential stem cell niche region the atrioventricular junction (AVj)3,4,5. Specifically, we sought to investigate whether global ischemia, caused by cardiac arrest, activate regenerative processes such as cardiomyocyte remodeling and cell renewal in the adult human heart.

When damage occur in cardiac tissue, the result is often an increase in fibrosis, hypertrophy, adipose tissue infiltration, lipofuscin accumulation or nuclei fragmentation. We expected histological changes in the cardiac arrest group, but no clear difference was observed by the systematic analysis of the tissue. The time window, 24days between the cardiac arrest and organ donation, seems too short for a histological remodeling of the normal myocardium to be demonstrated.

To further investigate potential differences between the two groups, the expression of cTnT, a part of the sarcomeres of cardiomyocytes, was studied. As expected, an even distribution was found in LV from the control group. In contrast, after cardiac arrest individual cardiomyocytes with a decreased expression of cTnT were discovered. Moreover, our results show that these cardiomyocytes with low cTnT upregulated the expression of the stem cell associated biomarkers. This is an interesting similarity to transient cardiomyocyte dedifferentiation, which is characterized by structural remodelling of the sarcomeres including decreased levels of cTnT7,9. We have previously reported that MDR1, SSEA4 and WT1 are expressed in small immature myocytes in the suggested hypoxic stem cell niche in the human AVj but not in LV from the same hearts4. Upregulation of these early cardiac biomarkers in a subpopulation of LV cardiomyocytes post ischemia supports the idea of dedifferentiation in existing cardiomyocytes. Furthermore, a study using brief non-lethal myocardial ischemiareperfusion model in sheep reported that MDR1 was upregulated both after 3 and 48h following reperfusion23. The authors proposed MDR1 as an early biomarker whose activation plays a pivotal role for cell survival. The expression of MDR1 in human LV following reperfusion after cardiac arrest in our study is consistent with a prolonged expression in response to global ischemia.

In addition, we found expression of the early cardiac transcription factor NKX2.5 in the LV cardiomyocytes with reduced cTnT interpreted as a potential reprogramming of adult cardiomyocytes into a more immature phenotype. NKX2.5 expression was detected in dedifferentiating rat cardiomyocytes in culture9 and is known to lie upstream of many essential genes for heart development13. Furthermore, in zebrafish, it was shown that activation of NKX2.5 was required for not only adult myocardial repair but also to provoke the associated proteolytic pathways of sarcomere disassembly as well as the proliferative response for cardiomyocyte renewal24. In line with this background, we suggest that the increase in NKX2.5 in combination with a decrease in cTnT expression may signify a remodeling process. Collectively, a reduction in cTnT and upregulation of stem cell associated biomarkers after an episode of global ischemia followed by few days of oxygen supply indicate a remodeling of the LV cardiomyocytes.

Intriguingly, hypoxia has been shown to induce dedifferentiation of early committed cells into pluripotency25. The fact that no nuclear Hif1 expression was observed in the LV after cardiac arrest despite the increased MDR1 expression after 14days of reperfusion is likely due to the very short half-life (~58min) of Hif1 after return to normal oxygen levels26,27. Under normal conditions Hif1 is expressed in the cytoplasm18, but when the oxygen levels drop Hif1 is instead accumulated in the nuclei17. Beyond its function as a transcriptional regulator for the cellular response to hypoxia, Hif1 plays a role in the activation of genes related to tissue repair28. In contrast to the findings in LV, nuclear Hif1 expression was detected in non-cardiomyocytes in the AVj in both groups, as previously reported4,5. This indicates that the AVj region holds a lower oxygen level than other parts of the heart, strengthening the theory of hypoxic stem cell niche in adult human heart. A hypoxia-responsive element has been identified in the early cardiac transcription factor WT1 sequence that bounds to Hif1 which was required for activation of the WT1 promotor29. WT1 has been correlated to epicardial regeneration30 as well as expression by endothelial cells31. We have previously reported that WT1 is expressed in the human AVj but not in LV cardiomyocytes4. In the present study, the co-expression of WT1 was found in small SSEA4+/cTnT+ myocytes in AVj. The numbers of WT1+/cTnT+ cells increased in the AVj after cardiac arrest, interpreted as a regenerative response to global hypoxia in the niche region. Another observation was increased numbers of WT1+/cTnT cells in LV after cardiac arrest (data not shown) interpreted as an activation of non-myocytes.

It is common for cardiomyocytes to have more than one nucleus. The nuclei are separated from each other in cardiomyocytes. The analyses of PCM1 expression revealed twin nuclei in cardiomyocytes. A systematic quantification of multiple large images showed that the number of twin nuclei increased after cardiac arrest, in both locations. The highest numbers were counted in the LV. Donor 21 was an outlier showing highest number of twin nuclei after the longest period of hypoxia (75min) compared to the others (Suppl. Table 1). However, it is difficult to draw conclusions from only one case. Binucleation takes place during the fetal development32. The absence of Ki67 or PCNA expression in the twin nuclei in the LV suggest that the results represent binucleation rather that proliferation. However, it should be noted that the half-lives of these two proliferation markers are short (~1 and 8h respectively)20,21. Although we cannot solidly determine whether the twin nuclei represent ongoing cell division or binucleation, it is worth noting that both these processes reflect mitosis33,34. Furthermore, it has been shown that PCM1 is a centrosome protein which localizes to the nuclear membrane2 and more specifically to dense structures on the cytoplasmic site of the nuclear envelope35. Therefore, the appearance of the PCM1 staining in the twin nuclei with two visible nuclear envelopes (see Fig.5b2,c2,d) is in itself evidence which strongly suggests binucleation rather than polyploidy within a single nucleus. In line with our results, double nuclei were observed in dedifferentiating cardiomyocytes days after apical resection in newborn mice, whereas neighbouring myocytes which did not undergo dedifferentiation or associated sarcomeric disorganization only displayed single nuclei36. Thus, regardless of whether they were destined for cell division or binucleation, the twin nuclei are consistent with a remodeling process.

Neither of the proliferation markers were found in cardiomyocytes in the LV after cardiac arrest, not even in the cardiomyocytes with the low cTnT expression suggesting that remodeling is a longer process, and that proliferation has not been initiated 14days following cardiac arrest. Support for this can be found in the study by Meckert et al. who found 12% of the LV myocytes contained Ki67+ nuclei in 713days-old infarcts. Earlier (16days) and also later (1421days), the portion of Ki67+ myocytes was significantly lower37. The absence of Ki67+ nuclei in the LV in the present study (14days after cardiac arrest) therefore seems to be largely in agreement with these results.

In contrast to the LV, PCNA and Ki67 were co-expressed with cardiac specific nuclei marker PCM1 in AVj, which may indicate increased proliferation in small myocytes after a period of global hypoxia. Ki67 has a shorter half-time than PCNA20,21, which could be an explanation to why more of PCNA+/PCM1+ nuclei compared to Ki67+/PCM1+ nuclei were detected. Another possibility is that PCNA can also be involved in DNA repair, including in human cardiomyocytes37. As there were clear examples of PCNA+/PCM1+ as well as Ki67+/PCM1+ twin nuclei in AVj, it appears that at least some of the PCNA positivity was associated with nuclei which had entered the cell cycle. Previously, we reported increased numbers of BrdU+ proliferating cells in the AVj using physical exercise in the adult rats3. In addition we have shown expression of biomarkers related to hypoxia, cardiac stem cells, proliferation and migration in the left and right AVj4,5 indicating that this region is of importance to cardiomyocyte cell renewal in human. I the current study, the increased expression of proliferation markers in the AVj after cardiac arrest suggests that more cardiomyocytes might had entered the cell cycle.

What may be the ultimate fates of the PCM1+ cardiomyocytes in AVj that displayed cell cycle markers? Regarding some of the PCM1+ nuclei that displayed no clear PCM1+ nuclear envelopes (Fig.4a2,a3), these are admittedly difficult to interpret. However, there is evidence to suggest that the insoluble perinuclear matrix remains in most phases of the cell cycle but disassembles only in pro-metaphase and metaphase of mitosis, making it possible to visualize myocyte nuclei almost throughout the whole cell cycle38. It thus seems possible that some of the Ki67+/PCM1+ and PCNA+/PCM1+ nuclei in the AVj in the present study were in prometaphase and metaphase.

In a study on infarcted human hearts, a low number of Ki67+ myocytes in the periinfarct zone had appearances consistent with conventional mitosis37. Thus, there is a slight possibility that minor portion of the Ki67+/PCM1+ and PCNA+/PCM1+ nuclei in the AVj may represent conventional cell division. However, Meckert et al. reported evidence to suggest that in human infarcts, entrance of cardiomyocytes into the cell cycle is transient and that endomitosis, leading to polyploidy rather than mitosis, is the final fate of cycling cells37. Nevertheless, since cardiac arrest and myocardial infarction are different conditions, there is a clear need for further studies into these issues. A possible explanation behind the differences between the AVj and the LV in the present study may be that the cardiomyocytes in the AVj are younger and in a more immature stage and thus perhaps able to express proliferation markers early after global ischemia.

Some limitations of the present study should be acknowledged. Immunohistochemistry data shows only a snapshot in time but provide important insights on co-expression of biomarkers in human adult cardiomyocytes. The low number of individuals and the limited range of the reperfusion period after cardiac arrest, as well as the short half-life of the chosen proliferation markers, makes it challenging to ascertain whether the twin nuclei were destined for binucleation, polyploidization or cell division. Also, some of the Ki67 and PCNA positivity may have been reflective of polyploidization and/or DNA damage, both of which may have occurred to varying extents. The methods and markers that we used did not allow us to investigate whether this was indeed the case. The physiological significance of the increased number of twin-nuclei as well as the Ki67+/PCM1+ and PCNA+/PCM1+ nuclei in and the remodelling cardiomyocytes after cardiac arrest thus needs further investigation. Nevertheless, the material is highly unique and may provide important insights into cellular response to cardiac arrest in human heart and clues for therapies aimed at improving heart regeneration.

The rest is here:
Global ischemia induces stemness and dedifferentiation in human adult cardiomyocytes after cardiac arrest | Scientific ... - Nature.com

Cardiac Disease Stem Cell Therapy Market

Understanding the segments helps in identifying the importance of different factors that aid market growth. This report gives you a clear vision of how the research is derived through primary and secondary sources considering expert opinion, patent analysis, the latest market development activity, and other influencing factors. The report throws light on the competitive landscape, segmentation, geographical expansion, and revenue, production, and consumption growth of the Cardiac Disease Stem Cell Therapy market.

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Some of the key players profiled in the study are:

Cellino Biotech Mesoblast BioCardia Cedars-Sinai Stem Cells Transplant Institute CCTRN Help Therapeutics Co., Ltd. Beijing Cellapy biotechnology Co., LTD CardioCell

Cardiac Disease Stem Cell Therapy Market Segmentation:

By Types:

Autologous Allogeneic

By Applications:

Preclinical Clinical Phase 1,2

Research Methodology:

This includes the collection of information through analysts to have them studied and filtered thoroughly in an try to provide good-sized predictions approximately the marketplace over the evaluate length. The research method further consists of interviews with main market influencers, which makes the primary research applicable and realistic. The secondary methods give a direct peek into the demand and deliver the connection. The market methodologies followed within the record offer specific facts analysis and provide a tour of the whole marketplace.

Market Geography:

The Cardiac Disease Stem Cell Therapy Market provides a diverse geographical landscape, with several areas exhibiting unique market characteristics. While some locations see rapid growth due to factors like economic expansion and technical advancements, other regions may experience slower but more consistent market expansion. Market trends vary greatly throughout regions due to factors including cultural influences, legal frameworks, and population developments. Businesses looking to go global must understand these regional differences and adjust their strategy to take advantage of local opportunities.

North America (U.S., Canada, China)

Europe (Germany, U.K., France, Italy, Russia, Spain, Rest of Europe)

Asia-Pacific (Japan, South Korea, China Taiwan, Southeast Asia, India)

Middle East, Africa, Latin America (Brazil, Mexico, Turkey, Israel, GCC Countries)

Informational Takeaways from the Market Study:

The Cardiac Disease Stem Cell Therapy Market report matches the completely examined and evaluated data of the noticeable companies and their situation in the market considering the impact of Coronavirus. The measured tools including SWOT analysis, Porter's five analysis, and assumption return debt were utilized while separating the improvement of the key players performing in the market.

If you are involved in the Cardiac Disease Stem Cell Therapy market or aim to be, then this study will provide you inclusive point of view. You must keep your market knowledge up to date segmented by top players. If you have a different set of players/manufacturers according to regional or countrywide segmented report we can provide customization according to your requirements.

Objectives of the Report:

To carefully analyze and forecast the market size by value and volume. To estimate the market shares of major segments. To showcase the development of the market in regions. To analyze micro-markets in terms of their contributions to the Cardiac Disease Stem Cell Therapy market, their prospects, and individual growth trends. To offer precise and useful details about factors affecting the growth of the market. To provide a meticulous assessment of crucial business strategies used by leading companies operating in the Cardiac Disease Stem Cell Therapy market, which include research and development, collaborations, agreements, partnerships, acquisitions, mergers, new developments, and product launches.

Key Questions Answered with this Study:

1) What makes Cardiac Disease Stem Cell Therapy Market feasible for long-term investment? 2) Teritorry that may see a steep rise in CAGR & Y-O-Y growth? 3) Which regions would have better demand for products/services? 4) What opportunity emerging regions would offer to established and new entrants in the market? 5) Risk side analysis connected with service providers? 6) How influencing are factors driving the demand for Cardiac Disease Stem Cell Therapy in the next few years? 7) What is the impact analysis of various factors in the global market growth? 8) What strategies of big players help them acquire a share in a mature market?

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Reasons to Purchase Report:

This report provides an in-depth exploration of the trends, challenges, and opportunities within this dynamic market landscape. From enabling technological advancements to driving innovation and sustainability initiatives, the Cardiac Disease Stem Cell Therapy market plays a pivotal role in shaping the modern world economy. The report throws light on the competitive landscape, segmentation, geographical expansion, and revenue, production, and consumption growth of the Cardiac Disease Stem Cell Therapy market. This report provides future products, joint ventures, marketing strategy, developments, mergers and acquisitions, marketing, promotions, revenue, import, export, CAGR values, the industry as a whole, and the particular competitors faced are also studied in the large-scale market. The report also analyzes the different segments along with major geographies that have more market demand.

Thanks for reading this article; you can also get individual chapter-wise sections or region-wise report versions.

Author Bio:

Money Singh is a seasoned content writer with over four years of experience in the market research sector. Her expertise spans various industries, including food and beverages, biotechnology, chemical and materials, defense and aerospace, consumer goods, etc. (https://www.linkedin.com/in/money-singh-590844163)

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Cardiac Disease Stem Cell Therapy Market 2024-2031: Emerging Trends, Growth Opportunities, Growth And Business ... - openPR

Welcome to the weekly edition of Healthcare IT Today Bonus Features. This article will be a weekly roundup of interesting stories, product announcements, new hires, partnerships, research studies, awards, sales, and more. Because theres so much happening out there in healthcare IT we arent able to cover in our full articles, we still want to make sure youre informed of all the latest news, announcements, and stories happening to help you better do your job.

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If you have news that youd like us to consider for a future edition ofHealthcare IT Today Bonus Features, please submit them onthis page. Please include any relevant links and let us know if news is under embargo. Note that submissions received after the close of business on Thursday may not be included in Bonus Features until the following week.

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Bonus Features June 16, 2024 40% of patients are willing to follow medical advice generated by AI, genetic testing ... - Healthcare IT Today

Genetics discoveries are rapidly identifying the people at greater risk for cancers and other diseases, but researchers at HealthPartners are concerned that they are happening too fast for doctors and patients to keep up.

The Bloomington-based health care provider is testing a new alert system to close that knowledge gap by identifying patients with inherited risks for diseases and automatically alerting their doctors of their need for testing. Screening guidelines changed three times in the past 18 months, just for breast and ovarian cancers, after researchers identified genetic variants that increased risks of those diseases, said Dr. Patrick O'Connor, a senior researcher with HealthPartners Institute.

"There is a risk of information overload," O'Connor said. "That's why we're creating a system to help organize these data in a way that's clear to patients so they can make informed decisions about treatment options that may be of benefit to them."

Using a $3.2 million federal grant announced earlier this week, HealthPartners hopes its alert system will hasten testing, which in turn will hasten diagnosis and treatment of diseases before they become severe or fatal. The goal is precision medicine tailoring treatments based on patients' unique circumstances but O'Connor said there are many examples in health care now when that approach is underused.

Research has identified numerous genetic variants that affect the course of type 2 diabetes, for instance, but those nuances aren't widely used right now to customize treatments, O'Connor said. Antidepressants are dispensed at a common starter dosage without consideration of the known variants that can dictate how well they will work, he added.

HealthPartners' study will identify patients at 40 clinics in Minnesota and western Wisconsin who haven't followed up on genetic testing results, even though they have one of seven variants linked to elevated risks of breast, colon or ovarian cancers. Doctors at 20 of those clinics will be prompted by the new alert system to talk with these patients about recommended tests or treatments. Researchers expect that these patients will receive more recommended screenings over the next three years than a comparison group of patients at the 20 other clinics that aren't receiving alerts.

Genes are chemical strands that program the body's cells based on hereditary information passed down from parents to children. Millions of variations alter how genes work in the body, but a much smaller portion has been linked through research so far to elevated rates of disease.

The Centers for Disease Control and Prevention recommends testing for 11 genetic variants that have proven links to cancer or heart disease, including the variants of the BRCA1 and BRCA2 genes that are closely tied to breast cancer. The American College of Medical Genetics and Genomics lists 81 variants that should be reported to doctors and patients because of their links to treatable conditions.

If HealthPartners' system works, it will be expanded to alert patients to other clinically important variants. Some only cause minute shifts in disease risks, while one known variation can increase lifetime risk of ovarian cancer from 1% to 70%. Another can identify smokers who have the best chance of reducing heart attack risks if they quit their habits.

"The difference can be gigantic in some cases," O'Connor said.

Deenya Craig, 52, of Maple Grove didn't hesitate when testing identified a BRCA2 mutation that increased her cancer risks and explained her family's tragic history with the disease. One cousin recently died from prostate cancer while another struggled with an aggressive breast cancer. The result "opened doors that previously had been closed," she said, including consultations with cancer specialists and insurance coverage of preventive treatments. She had a mastectomy last year to remove breast tissue that posed cancer risks, and had a gynecological procedure this month to reduce her risks for ovarian cancer.

Craig spread the word of her results, and now her sisters and three of her six children have been tested. Her whole genome testing of thousands of genes at once produced other interesting information about her susceptibility to caffeine and sleep disturbances, but mostly she said she felt empowered by the knowledge about her cancer risks.

"It gives the control back to you over your health instead of sitting and wondering what, if or when," Craig said.

Craig received free testing through HealthPartners' partnership with California-based Helix Inc. to gather genetic data from 100,000 volunteers. The myGenetics program is designed to alert volunteers to health risks but also amass genetic information for further identification of troublesome variants. Out of 40,000 volunteers, testing has found more than 600 with inherited risks for breast, colon or ovarian cancers.

Mayo Clinic will soon publish interim data from a similar project called Tapestry, which is recruiting 100,000 volunteers from its campuses in Minnesota, Arizona and Florida to identify those with any of the 11 CDC-identified variants. Results will show how many people with these variants wouldn't have qualified for screening under current insurance and health system guidelines, said Dr. Jewel Samadder, co-director of precision oncology at Mayo Clinic's cancer center in Arizona.

Mayo also will be following 15,000 of these patients over five years to compare their health and health care spending with patients who don't have any of the variants. Screening just for the breast cancer variants used to cost $5,000, but now tests of thousands of genes at once cost around $500. Samadder said health systems will need to expand to account for a new generation of patients who are acting on this affordable genetic information.

Some studies already estimate that the cost is low enough to warrant widespread testing because it will end up saving money by identifying cancers before they require high-cost treatments.

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Genetic discoveries map out your health risks, if you can keep up with them - Star Tribune

Data protection watchdogs in Canada and the UK have announced a joint investigation into a data breach at genetics testing company 23andMe that affected seven million customers in 2023.

Since 2006, San Francisco-based genetic testing company 23andMe has sold over 12 million DNA testing kits. It analyses customers saliva to provide insights into health and ancestry.

In October 2023, 23andMe reported a security breach in which hackers gained access to the personal information of millions of customers by using old passwords. In some cases, the information accessed included family trees, birth years and geographic locations.

The Information Commissioner's Office (ICO) in the UK and the Office of the Privacy Commissioner of Canada (OPC) will examine the scope of information that was exposed by the breach and potential harms to affected people.

John Edwards, the UK information commissioner, said: People need to trust that any organisation handling their most sensitive personal information has the appropriate security and safeguards in place.

He added: This data breach had an international impact, and we look forward to collaborating with our Canadian counterparts to ensure the personal information of people in the UK is protected.

The strength of 23andMes safeguards to protect the information within its control will also be investigated, as well as whether the company provided adequate notification about the breach to the two regulators and affected people.

Philippe Dufresne, the Canadian privacy commissioner, said: In the wrong hands, an individuals genetic information could be misused for surveillance or discrimination. Ensuring that personal information is adequately protected against attacks by malicious actors is an important focus for privacy authorities in Canada and around the world.

According to the Guardian, a 23andMe spokesperson had previously said that the company did not detect a breach within its systems and instead attributed the incident to compromised recycled login credentials from certain users.

However, 23andMe has said it will cooperate with the investigation and the regulators reasonable requests.

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Genetics testing company 23andMe to be probed over a data breach that affected 7 million users - E&T Magazine

Around the world, hundreds of people have had their bodies frozen at extremely low temperatures, just after death in the hope that they can be revived in the future.

Some are so confident that theyll wake up in the future that their loved ones have left them voicemail messages.

But the moment when people can be revived by science could come sooner than we expect, according to Dennis Kowalski of Michigans Cryonics Institute.

MORE: Woman in her 30s kidnapped and gang raped in broad daylight at Essex graveyard MORE: Couple held after 13 children found chained to their beds in California home

Kowalski told the Daily Star, If you take something like CPR, that would have seemed unbelievable 100 years ago. Now we take that technology for granted.

Cryonically bringing someone back to life should definitely be doable in 100 years, but it could be as soon as ten.

Companies pump peoples brains full of cryoprotectant fluid before being frozen in the hope the brains will last decades or even hundreds of years.

Many cryonics fans have their heads frozen not their whole bodies imagining that in the future, brain transplants will be possible.

Kowalski says that innovations in technologies such as stem cells may make it possible to revive frozen bodies at some point in the future.

Another tech company, Humai is monitoring developments in robotics, medical treatments and believes people will come back from the dead within 30 years.

The company believes that within three decades, technology will have advanced so that people can freeze their brains then have them transplanted into an artificial, robot-like body after death.

CEO Josh Bocanegra told Popular Science, Well first collect extensive data on our members for years prior to their death via various apps were developing.

After death well freeze the brain using cryonics technology. When the technology is fully developed well implant the brain into an artificial body. The artificial body functions will be controlled with your thoughts by measuring brain waves.

As the brain ages well use nanotechnology to repair and improve cells. Cloning technology is going to help with this too.

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Humans frozen by cryogenics 'could be brought back to life in 10 years' - Yahoo Canada Shine On

Ive had many conversations over the years about life extension, and the idea often meets resistance. People become upset when they hear of an individual whose life has been cut short by a disease, yet when confronted with the possibility of generally extending all human life, they react negatively. Life is too difficult to contemplate going on indefinitely is a common response. But people generally do not want to end their lives at any point unless they are in enormous painphysically, mentally, or spiritually. And if they were to absorb the ongoing improvements of life in all its dimensions, most such afflictions would be alleviated. That is, extending human life would also mean vastly improving it.

But how will nanotechnology actually make this possible? In my view, the long-term goal is medical nanorobots. These will be made from diamondoid parts with onboard sensors, manipulators, computers, communicators, and possibly power supplies. It is intuitive to imagine nanobots as tiny metal robotic submarines chugging through the bloodstream, but physics at the nanoscale requires a substantially different approach. At this scale, water is a powerful solvent, and oxidant molecules are highly reactive, so strong materials like diamondoid will be needed.

And whereas macro-scale submarines can smoothly propel themselves through liquids, for nanoscale objects, fluid dynamics are dominated by sticky frictional forces. Imagine trying to swim through peanut butter! So nanobots will need to harness different principles of propulsion. Likewise, nanobots probably wont be able to store enough onboard energy or computing power to accomplish all their tasks independently, so they will need to be designed to draw energy from their surroundings and either obey outside control signals or collaborate with one another to do computation.

To maintain our bodies and otherwise counteract health problems, we will all need a huge number of nanobots, each about the size of a cell. The best available estimates say that the human body is made of several tens of trillions of biological cells. If we augment ourselves with just 1 nanobot per 100 cells, this would amount to several hundred billion nanobots. It remains to be seen, though, what ratio is optimal. It might turn out, for example, that advanced nanobots could be effective even at a cell-to-nanobot ratio several orders of magnitude greater.

One of the main effects of aging is degrading organ performance, so a key role of these nanobots will be to repair and augment them. Other than expanding our neocortex, this will mainly involve helping our nonsensory organs to efficiently place substances into the blood supply (or lymph system) or remove them. By monitoring the supply of these vital substances, adjusting their levels as needed, and maintaining organ structures, nanobots can keep a persons body in good health indefinitely. Ultimately, nanobots will be able to replace biological organs altogether, if needed or desired.

But nanobots wont be limited to preserving the bodys normal function. They could also be used to adjust concentrations of various substances in our blood to levels more optimal than what would normally occur in the body. Hormones could be tweaked to give us more energy and focus, or speed up the bodys natural healing and repair. If optimizing hormones could make our sleep more efficient, it would in effect be backdoor life extension. If you just go from needing eight hours of sleep a night to seven hours, that adds as much waking existence to the average life as five more years of lifespan!

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The Secret to Living Past 120 Years Old? Nanobots - WIRED

Study design

A dual-center, retrospective study of patients undergoing HCT for SAA was conducted at Vanderbilt University Medical Center (VUMC) and the associated Veterans Affairs hospital, Tennessee Valley Healthcare System (TVHS). The VUMC and TVHS Institutional Review Board approved the study. Patients with a SAA diagnosis who underwent first allogeneic HCT using FCR conditioning regimen at VUMC or TVHS between January 2016 and May 2022 were included in the study. Patients were excluded if they were younger than 18 or had not completed all planned treatments at the time of data collection.

All patients received conditioning per established protocol as determined by degree of HLA-matching with their designated donor (Fig.1). Patients received PBSC or BM grafts per treating physicians discretion with allogeneic HCT performed on day 0. In patients with matched related, matched unrelated, or 1-allele mismatched donors, fludarabine (30mg/m2) was given intravenously for four days (7 to 4, i.e., 7 to 4 days before transplantation) in combination with cyclophosphamide (750mg/m2) given intravenously for three days (6 to 4) and anti-thymocyte globulin (rabbit) (3.75mg/kg) for two days (2 and 1). They were also given rituximab (375mg/m2) on days 13, 7, +1, and +8.

Doses and timing of each agent in FCR conditioning regimens for patients undergoing HCT for SAA with matched related, matched unrelated, or 1-allele mismatch donor (a) or haploidentical donor (b). Time in days is represented along the horizontal axis progressing from left to right, with day of transplantation depicted as day 0.

Patients who underwent HCT from a haploidentical donor received fludarabine (30mg/m2) intravenously for five days (6 to 2) and cyclophosphamide (14.5mg/kg) for two days (6 to 5). They also received rituximab (200mg/m2) on day +5. All patients received total body irradiation at a dose of 200cGy on day 0 for those with matched related, unrelated, or 1-allele mismatched donors, or on day 1 for those with haploidentical donors.

Prophylaxis for GVHD in patients with matched related, matched unrelated, or 1-allele mismatch donors consisted of tacrolimus starting on day 3 and methotrexate, 5mg/m2 intravenously, on days +1, +3, and +6 after transplantation. Patients with haploidentical donors received standard post-transplant cyclophosphamide (50mg/kg on days +3 and +4), and tacrolimus and mycophenolate mofetil starting on day +5. Tacrolimus levels were adjusted to a goal range of 515ng/mL per institutional standard, and administered for 180 days, at which point a taper was initiated provided absence of GVHD. Mycophenolate mofetil was administered until day +35 in patients with haploidentical donors.

The primary outcome of interest was GVHD-free/relapse-free survival. A patient was considered to have an event if they experienced moderate or severe GVHD (including both acute GVHD [aGVHD] and cGVHD), relapse, or death. If a patient experienced multiple events, the earliest event date was used as the time to event (e.g., if a patient had a diagnosis of both aGVHD and cGVHD, the earliest date of diagnosis was used). If a patient did not experience an event until the end of follow-up time (i.e., the last date the patient was seen in the clinic or lost to follow-up [unable to contact, transitioned care to another city, etc.]), it was censored. Acute and chronic GVHD were graded according to Glucksburg and 2014 National Institutes of Health consensus criteria, respectively [17, 18]. Secondary outcomes included time to engraftment, incidence of graft failure, incidence of GVHD, rate of viral reactivation, post-HCT disease status, and number of inpatient hospital days.

Descriptive statistics were used to summarize the patient characteristics. Medians and interquartile ranges (IQRs) were used for continuous variables, while frequencies and percentages were used for categorical variables. Differences in patient characteristics were tested for using Wilcoxon rank sum tests for continuous variables and chi-square tests for categorical variables. Probability of GRFS over time was estimated using the Kaplan-Meier estimation method.

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Efficacy and safety of outpatient fludarabine, cyclophosphamide, and rituximab based allogeneic hematopoietic cell ... - Nature.com

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Salute to saving a life: Local Air Force man answers the call for stem cell donation - Citrus County Chronicle

Key Takeaways

CAR-T cell therapy doesnt increase a persons risk of secondary cancers, researchers reportThe risk of a secondary cancer is slightly more than 6%, about the same as stem cell transplantation to treat blood cancersOther factors appear to be involved in secondary cancers

THURSDAY, June 13, 2024 (HealthDay News) CAR-T cell therapy to treat blood cancers is safer than previously thought, with little risk that the immunotherapy will create secondary cancers, a new study finds.

The U.S. Food and Drug Administration issued a warning in November 2023 about a risk of secondary cancers that might be associated with CAR-T cell therapy.

But a study of more than 700 patients treated at Stanford University found that the risk was just over 6% in the three years after a cancer patient received CAR-T cell immunotherapy, researchers reported June 13 in the New England Journal of Medicine.

That risk is roughly similar to that of patients who receive stem cell transplants rather than CAR-T cell therapy to treat their blood cancers, researchers said.

These are lifesaving therapies that come with a very low risk of secondary cancers. The challenge lies in how to predict which patients are at higher risk, and why, said researcher Dr. Ash Alizadeh, a professor of medicine at Stanford.

In CAR-T cell therapy, immune cells called T-cells are harvested from a patient and genetically engineered to more efficiently seek out and kill cancer cells.

This therapy typically is used to treat blood cancers like leukemia, lymphoma and multiple myeloma, according to the American Cancer Society.

But one concern is that if the genetic engineering is imprecise, the T-cells meant to attack a persons cancer might instead become cancerous themselves.

To see whether this risk is real, the research team analyzed data drawn from Stanford Medicines large bank of tissue and blood samples from people receiving CAR-T cell therapy.

They found no evidence that the T-cells responsible for some patients secondary cancers were the T-cells that had been engineered for CAR-T cell therapy. The T-cells were distinct on both genetic and molecular levels.

But in one patient who rapidly developed and died from a T-cell lymphoma, researchers found a clue that could explain why secondary cancers sometimes happen.

Both sets of T-cells in that patient the CAR-T cells and the T-cells responsible for the secondary cancer had been infected with a virus known to play a role in cancer development. The patient also had a history of autoimmune disease prior to a cancer diagnosis.

We compared protein levels, RNA sequences and DNA from single cells across multiple tissues and time points to determine that the therapy didnt introduce the lymphoma into this patient; instead it was already brewing in their body at very low levels, Alizadeh said in a Stanford news release.

This suggests that secondary cancers might be prompted by chemotherapy done prior to CAR-T cell therapy, which suppresses a persons immune response to such viruses, researchers said. They also might be due to some other side effect from the treatment, rather than genetic engineering gone wrong.

These results may help researchers focus on the immune suppression that can precede and often follows CAR-T cell therapy, said researcher Dr. David Miklos, chief of bone marrow transplantation and cellular therapy at Stanford Medicine.

More information

The American Cancer Society has more about CAR-T cell therapy.

SOURCE: Stanford University, news release, June 12, 2024

What This Means For You

People with blood cancers can receive CAR-T cell therapy with little fear of increased risk for secondary cancers, researchers say.

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Secondary Tumors After CAR-T Cancer Therapies Are Rare: Study | Fox 11 Tri Cities Fox 41 Yakima - FOX 11 and FOX 41

DADDY, can we play football?

Those are the most beautiful words in the world to Mo Hussain, 38, from Blackburn, because they mean his five-year-old son Eesa is having a good day.

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And to ensure more good days lie ahead, this devoted father is a man on a mission, seeking the stem cell match that will save his little boys life.

On New Years Day, Eesa felt sick and looked really pale, Mo says. Concerned, we took him to A&E.

Its every parents worst nightmare. A few hours later their son was being transferred by ambulance to Royal Manchester Childrens Hospital.

Normal levels of haemoglobin a protein which transports oxygen around the body are 120-150g/l. Eesas was 42g/l due to dangerously low red blood cell levels.

I want to go home and play with my dinosaurs, Eesa cried. Mos heart broke.

Mo cuddled Eesa as he had a general anaesthetic for a bone marrow biopsy.

That night, the whole family Mo, his wife and two-year-old son Ali crammed into Eesas cubicle on ward 86 to sleep. We needed to be together, Mo says.

The biopsy showed Eesa had a rare, life-threatening condition called aplastic anaemia, meaning his bone marrow and stem cells dont produce enough red and white blood cells and platelets.

The best cure is a bone marrow transplant from a matching donor.

While preparations were made to test the familys suitability, Eesa had a Hickman line fitted in his chest so doctors could administer medicines and take blood.

To make it less frightening, I bought some plastic tubing and stuck it to my chest too, says Mo.

Back home after a week in hospital, the family were on lockdown Eesa off school and Mo and his wife on leave from work.

We cant risk Eesa catching something his bodys weak immune system cant fight, Mo says.

Devastatingly, no one in Eesas family is a donor match for him.

But there is hope. Fifty years ago, Shirley Nolan was so determined to save her son Anthonys life that she set up the worlds first stem cell register.

Since 1974, the charity Anthony Nolan has helped bring about more than 26,500 transplants for people around the world.

If youre from a minority ethnic background, youre more likely to have a rare or completely unique tissue type.

Thats why theres a pressing need to recruit more people from diverse backgrounds to the register to help patients like Eesa find the lifesaving matches they need.

I had to educate our community, Mo says. The team at Anthony Nolan sent me swabs for people to wipe inside their mouths and envelopes to post them back.

And, in February, we set up our first registration stall at a football tournament.

Since then, Mo and his family and friends have visited mosques, universities and football stadiums including the Etihad, Turf Moor and Ewood Park 40 locations in all, adding 1,200 potential new donors to the register.

Sadly, Eesa is still waiting for his match and remains dependant on blood transfusions every three weeks.

I find patience in the words of the Quran saving one life is like saving the whole of humanity, says Mo.

Anthony Nolan shares its register across the world so the people we sign up could save lives in Bangladesh, Pakistan anywhere.

For now, Eesa has good days watching Arsenal and racing his police cars and bad. In April, he was hospitalised because his Hickman line became infected.

Hes the bravest five-year-old in Britain, Mo says. But we just want a normal childhood for him.

The greatest Fathers Day gift I could receive is a match for Eesa. So Im appealing to other dads log on to anthonynolan.org today.

You could save someone like my sons life.

Follow the My Name is Eesa campaign on Instagram at @mynameiseesa

Joining the stem cell register is easy. You must be aged between 16 and 30, as research shows younger donors offer better survival rates for patients.

Fill in a form at anthonynolan.org to receive a swab pack then take a sample and send it back.

Ken (above), 26, from Tower Hamlets, signed up to the register eight years ago at an Anthony Nolan stand handing out Krispy Kreme doughnuts.

And three years later he discovered he was a match for someone.

I was given G-CSF injections at home for four days, he says. G-CSF injections boost white cells and release stem cells into the bloodstream ready to collect.

They gave me minor headaches and muscle pain nothing more. On the fifth day I was in hospital, donating my stem cells and afterwards I was fine.

I hope that more people will join the stem cell register and help Anthony Nolan save the lives of people with blood cancer and blood disorders.

If a family member or friend was diagnosed with blood cancer, it would make such a difference to know that they have a match and a second chance at life.

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Are you under 30? Join the stem cell register and be part of a one million strong team of lifesavers. Sign up at anthonynolan.org

If youre over 30 you can still save lives. It costs Anthony Nolan 40 to recruit each lifesaver to the register, so please support with a gift now!

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Dad urges recruits to sign up in bid to save young son with rare blood condition... - The Irish Sun

Vertex Pharmaceuticals Incorporated (Nasdaq: VRTX) today announced longer-term data for CASGEVY (exagamglogene autotemcel [exa-cel]) from global clinical trials in people with severe sickle cell disease (SCD) or transfusion-dependent beta thalassemia (TDT). The results, presented at the annual European Hematology Association (EHA) Congress, confirm the transformative, consistent and durable clinical benefits of CASGEVY over time. CASGEVY is the first and only approved CRISPR-based gene-editing therapy.

The data being presented are from more than 100 patients (46 SCD; 56 TDT) treated with exa-cel in clinical trials, with the longest follow-up now extending more than 5 years. The efficacy results are consistent with the previously reported primary and key secondary endpoints analyses from these exa-cel studies and continue to demonstrate transformative clinical benefit with durable and stable levels of fetal hemoglobin (HbF) and allelic editing.

The transformative benefit seen in patients with sickle cell disease in the trial is impressive given the significant and cumulative burden of disease faced by people living with this blood disorder, said Haydar Frangoul, M.D., M.S., Medical Director of Pediatric Hematology and Oncology at Sarah Cannon Research Institute and HCA Healthcares TriStar Centennial Childrens Hospital. I am eager to offer this therapy and the opportunity of a potential functional cure to my eligible patients.

The comprehensive data set presented today for adult and adolescent TDT patients adds to the growing body of evidence for CASGEVY, and it is important to now ensure the availability of this innovative treatment to patients in the real world as soon as possible, said Franco Locatelli, M.D., Ph.D., Professor of Pediatrics at the Catholic University of the Sacred Heart of Rome, Director of the Department of Pediatric Hematology and Oncology at Bambino Ges Childrens Hospital. With the longest follow up now more than five years, alongside stable editing and sustained fetal hemoglobin levels, I have conviction in the durable benefit to the patients treated with CASGEVY.

New data presented from CASGEVY pivotal trials

In both SCD and TDT patients, edited levels of BCL11A alleles were stable over time in bone marrow and peripheral blood indicating successful editing in the long-term hematopoietic stem cells. All patients engrafted neutrophils and platelets after exa-cel infusion. The safety profile of exa-cel was generally consistent with myeloablative conditioning with busulfan and autologous hematopoietic stem cell transplant.

These longer-term data for CASGEVY from the CLIMB clinical trials will be shared as outlined below:

Vertex will also share five health economics abstracts at the EHA Congress.

About Sickle Cell Disease (SCD)

SCD is a debilitating, progressive, life shortening genetic disease. SCD patients report health-related quality of life scores well below the general population and significant health care resource utilization. SCD affects the red blood cells, which are essential for carrying oxygen to all organs and tissues of the body. SCD causes severe pain, organ damage and shortened life span due to misshapen or sickled red blood cells. The clinical hallmark of SCD is vaso-occlusive crises (VOCs), which are caused by blockages of blood vessels by sickled red blood cells and result in severe and debilitating pain that can happen anywhere in the body at any time. SCD requires lifelong treatment and significant use of health care resources, and ultimately results in reduced life expectancy, decreased quality of life and reduced lifetime earnings and productivity. In Europe, the mean age of death for patients living with SCD is around 40 years. Stem cell transplant from a matched donor is a potentially curative option but is only available to a small fraction of people living with SCD because of the lack of available donors.

About Transfusion-Dependent Beta Thalassemia (TDT)

TDT is a serious, life-threatening genetic disease. TDT patients report health-related quality of life scores below the general population and significant health care resource utilization. TDT requires frequent blood transfusions and iron chelation therapy throughout a persons life. Due to anemia, patients living with TDT may experience fatigue and shortness of breath, and infants may develop failure to thrive, jaundice and feeding problems. Complications of TDT can also include an enlarged spleen, liver and/or heart, misshapen bones and delayed puberty. TDT requires lifelong treatment and significant use of health care resources, and ultimately results in reduced life expectancy, decreased quality of life and reduced lifetime earnings and productivity. In Europe, the mean age of death for patients living with TDT is 50-55 years. Stem cell transplant from a matched donor is a potentially curative option but is only available to a small fraction of people living with TDT because of the lack of available donors.

About CASGEVY (exagamglogene autotemcel [exa-cel])

CASGEVY is a non-viral, ex vivo CRISPR/Cas9 gene-edited cell therapy for eligible patients with SCD or TDT, in which a patients own hematopoietic stem and progenitor cells are edited at the erythroid specific enhancer region of the BCL11A gene through a precise double-strand break. This edit results in the production of high levels of fetal hemoglobin (HbF; hemoglobin F) in red blood cells. HbF is the form of the oxygen-carrying hemoglobin that is naturally present during fetal development, which then switches to the adult form of hemoglobin after birth.

CASGEVY has been shown to reduce or eliminate VOCs for patients with SCD and transfusion requirements for patients with TDT.

CASGEVY is approved for certain indications in multiple jurisdictions for eligible patients.

About the CLIMB Studies

The ongoing Phase 1/2/3 open-label trials, CLIMB-111 and CLIMB-121, are designed to assess the safety and efficacy of a single dose of CASGEVY in patients ages 12 to 35 years with TDT or with SCD, characterized by recurrent VOCs, respectively. The trials are now closed for enrollment. Patients will be followed for approximately two years after CASGEVY infusion. Each patient will be asked to participate in the ongoing long-term, open-label trial, CLIMB-131. CLIMB-131 is designed to evaluate the safety and efficacy of CASGEVY in patients who received CASGEVY in other CLIMB studies. The trial is designed to follow patients for up to 15 years after CASGEVY infusion.

U.S. INDICATIONS AND IMPORTANT SAFETY INFORMATION FOR CASGEVY (exagamglogene autotemcel)

WHAT IS CASGEVY?

CASGEVY is a one-time therapy used to treat people aged 12 years and older with:

CASGEVY is made specifically for each patient, using the patients own edited blood stem cells, and increases the production of a special type of hemoglobin called hemoglobin F (fetal hemoglobin or HbF). Having more HbF increases overall hemoglobin levels and has been shown to improve the production and function of red blood cells. This can eliminate VOCs in people with sickle cell disease and eliminate the need for regular blood transfusions in people with beta thalassemia.

IMPORTANT SAFETY INFORMATION

What is the most important information I should know about CASGEVY?

After treatment with CASGEVY, you will have fewer blood cells for a while until CASGEVY takes hold (engrafts) into your bone marrow. This includes low levels of platelets (cells that usually help the blood to clot) and white blood cells (cells that usually fight infections). Your doctor will monitor this and give you treatment as required. The doctor will tell you when blood cell levels return to safe levels.

You may experience side effects associated with other medicines administered as part of the treatment regimen for CASGEVY. Talk to your physician regarding those possible side effects. Your healthcare provider may give you other medicines to treat your side effects.

How will I receive CASGEVY?

Your healthcare provider will give you other medicines, including a conditioning medicine, as part of your treatment with CASGEVY. Its important to talk to your healthcare provider about the risks and benefits of all medicines involved in your treatment.

After receiving the conditioning medicine, it may not be possible for you to become pregnant or father a child. You should discuss options for fertility preservation with your healthcare provider before treatment.

STEP 1: Before CASGEVY treatment, a doctor will give you mobilization medicine(s). This medicine moves blood stem cells from your bone marrow into the blood stream. The blood stem cells are then collected in a machine that separates the different blood cells (this is called apheresis). This entire process may happen more than once. Each time, it can take up to one week.

During this step rescue cells are also collected and stored at the hospital. These are your existing blood stem cells and are kept untreated just in case there is a problem in the treatment process. If CASGEVY cannot be given after the conditioning medicine, or if the modified blood stem cells do not take hold (engraft) in the body, these rescue cells will be given back to you. If you are given rescue cells, you will not have any treatment benefit from CASGEVY.

STEP 2: After they are collected, your blood stem cells will be sent to the manufacturing site where they are used to make CASGEVY. It may take up to 6 months from the time your cells are collected to manufacture and test CASGEVY before it is sent back to your healthcare provider.

STEP 3: Shortly before your stem cell transplant, your healthcare provider will give you a conditioning medicine for a few days in hospital. This will prepare you for treatment by clearing cells from the bone marrow, so they can be replaced with the modified cells in CASGEVY. After you are given this medicine, your blood cell levels will fall to very low levels. You will stay in the hospital for this step and remain in the hospital until after the infusion with CASGEVY.

STEP 4: One or more vials of CASGEVY will be given into a vein (intravenous infusion) over a short period of time.

After the CASGEVY infusion, you will stay in hospital so that your healthcare provider can closely monitor your recovery. This can take 4-6 weeks, but times can vary. Your healthcare provider will decide when you can go home.

What should I avoid after receiving CASGEVY?

What are the possible or reasonably likely side effects of CASGEVY?

The most common side effects of CASGEVY include:

Your healthcare provider will test your blood to check for low levels of blood cells (including platelets and white blood cells). Tell your healthcare provider right away if you get any of the following symptoms:

These are not all the possible side effects of CASGEVY. Call your doctor for medical advice about side effects. You may report side effects to FDA at 1-800-FDA-1088.

General information about the safe and effective use of CASGEVY

Talk to your healthcare provider about any health concerns.

Please see full Prescribing Information including Patient Information for CASGEVY.

About Vertex

Vertex is a global biotechnology company that invests in scientific innovation to create transformative medicines for people with serious diseases. The company has approved medicines that treat the underlying causes of multiple chronic, life-shortening genetic diseases cystic fibrosis, sickle cell disease and transfusion-dependent beta thalassemia and continues to advance clinical and research programs in these diseases. Vertex also has a robust clinical pipeline of investigational therapies across a range of modalities in other serious diseases where it has deep insight into causal human biology, including acute and neuropathic pain, APOL1-mediated kidney disease, IgA nephropathy, autosomal dominant polycystic kidney disease, type 1 diabetes, myotonic dystrophy type 1 and alpha-1 antitrypsin deficiency.

Vertex was founded in 1989 and has its global headquarters in Boston, with international headquarters in London. Additionally, the company has research and development sites and commercial offices in North America, Europe, Australia, Latin America and the Middle East. Vertex is consistently recognized as one of the industry's top places to work, including 14 consecutive years on Science magazine's Top Employers list and one of Fortunes 100 Best Companies to Work For. For company updates and to learn more about Vertex's history of innovation, visit http://www.vrtx.com or follow us on LinkedIn , YouTube and Twitter/X .

(VRTX-GEN)

Vertex Special Note Regarding Forward-Looking Statements

This press release contains forward-looking statements as defined in the Private Securities Litigation Reform Act of 1995, as amended, including, without limitation, the statements by Haydar Frangoul, M.D., M.S., and Franco Locatelli, M.D., Ph.D., in this press release, and statements regarding our expectations for and the anticipated benefits of CASGEVY, our plans to share longer-term data for CASGEVY from the CLIMB clinical trials and to share health economics abstracts at the EHA Congress, and our plans for and design of the CLIMB studies. While we believe the forward-looking statements contained in this press release are accurate, these forward-looking statements represent the company's beliefs only as of the date of this press release and there are a number of risks and uncertainties that could cause actual events or results to differ materially from those expressed or implied by such forward-looking statements. Those risks and uncertainties include, among other things, that data from the company's development programs may not support registration or further development of its compounds due to safety, efficacy, and other reasons, and other risks listed under the heading Risk Factors in Vertex's most recent annual report and subsequent quarterly reports filed with the Securities and Exchange Commission at http://www.sec.gov and available through the company's website at http://www.vrtx.com . You should not place undue reliance on these statements, or the scientific data presented. Vertex disclaims any obligation to update the information contained in this press release as new information becomes available.

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Media: mediainfo@vrtx.com or International: +44 20 3204 5275 or U.S.: 617-341-6992 or Heather Nichols: +1 617-839-3607

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Vertex Presents Positive Long-Term Data On CASGEVY (exagamglogene autotemcel) at the 2024 Annual European ... - Agenzia ANSA

There's an assumption that if you're a woman in a heterosexual relationship or marriage going through menopause, you'll have a partner who can step in. But that isn't always a guarantee.

Menopause can put "a huge strain" on relationships, Dr. Deirdre Forde - founder of Cile Medical in Athlone, the first dedicated Menopause Clinic in the Midlands - says, heterosexual or not.

"There's a lot of men who don't know about menopause, don't want to know about menopause and their woman, and their relationship becomes strained because she's turning into a crazy person and he doesn't know who she is anymore.

"And all of a sudden then they start to drift. So when they start to drift then, there's nothing to talk about anymore. Sure, a lot of them end up getting divorced."

"I was one of the one such person", Forde says, adding that she got divorced at 44, six years after starting menopause.

When she started menopause in her late 30s, Forde became gripped by what she calls an "irrational anxiety".

"I describe it like a washing machine, churning in your stomach, that just won't stop", says Forde. "I just wanted to detach my head for my body and make it stop."

Now, imagine you're a menopausal woman or a trans man in a relationship with another menopausal person. Who steps in to pick up the slack then?

Forde has opened up her Athlone practice to LGBT people going through menopause by organising free information events to offer support and advice, on 13 and 15 June.

"We need to talk about menopause as people who are going through menopause, as opposed to women going through menopause", she says. "I want to get rid of that stigma. There's nobody judging you. You are a person like everybody else, and you are going to go through this transition."

How we talk about the menopause has shifted radically in recent years, both globally and at home. A notable watershed moment in this shift was when scores of Irish women contacted Liveline with their often anguished accounts of menopause for five days straight, releasing years of suppressed pain.

Compounding this has been a glut of new research into the once mysterious part of a woman's life. This also backed up what many women had been saying in no uncertain terms: 55% of Irish women described menopause and perimenopause as a negative experience, compared to 22% calling it positive.

On top of that, an overwhelming majority of women (93%) agreed on menopause having a significant impact on a woman's life.

Menopause is defined as having had no period for 12 months, or if a woman has no ovaries due to surgery. It's caused by a drop in the production of key hormones estrogen, progesterone, and testosterone. It can span anywhere from four to 11 years and can bring on symptoms ranging from night sweats, mood swings and brain fog, to disrupted sleep, a burning sensation in the mouth and vaginal dryness.

Depending on the person, the stakes are undeniably high when it comes to untreated menopause, Forde says. Of course, there will always be some who practically sail through the experience, barely noticing a change. But the majority will undergo often seismic shifts in their physical and mental health. Research shows that fluctuating hormones can even have a lasting effect on the brain's structure.

"You're going to get women who just don't feel themselves anymore, who don't know who they are anymore. They feel that they can't work. They feel that they can't contribute to society anymore. Their brain is foggy and all of that. But they just feel worthless."

In some cases, those affected can become suicidal*, she says. People living with preexisting mental health conditions are particularly at risk.

"You've really got to watch a woman who, all of a sudden, is changing. Her mood is changing. Her mood swings are horrendous and her rage is terrible. She's not sleeping and she has uncontrollable anxiety. She's a menopausal woman and she needs help."

Finding the right support during menopause can still be challenging, particularly for LGBT people, she adds.

Although she herself hasn't treated any trans men presenting with menopause, she stresses that, "it is going to happen". "I am now 64, so I won't be doing this forever. But the new doctors coming on board who are hopefully going to be treating an awful lot of people with menopause, they're going to see it.

"I suppose trans people need to be aware. Trans men who are born with a uterus or who are born female, they need to be aware that they're going to face this in the future."

Forde's aim isn't to scaremonger, but to illuminate the reality of what many people experience. The hope is that in doing so, generations of people can prepare for the shift. There are already 600,000 menopausal women in Ireland, and by 2030 it is estimated that there will be 1.1 billion menopausal women on the planet.

Increased research and a greater understanding of hormone replacement therapy (HRT) and other treatments has made managing menopause considerably easier for many people.

Although a 1990s study - the Women's Health Initiative, which set out to examine the link between HRT and cardiovascular disease and studied the health of 160,000 postmenopausal women - skewed public perception against HRT, suggesting that it caused breast cancer, greater research has now shown that such risks are low for healthy women under the age of 60 and HRT is still a profoundly helpful treatment option for many women.

With the right treatment, Forde says, "You've got the benefit in that you've got your quality of life back. You're able to function as a person in this society. You're able to work, you're able to be happy."

To register for the free sessions on Saturday 15 June at 11am and 3pm email info@ceilemedical.ie with your name and age.

*If you are affected by any of the issues raised in this article, you can contact The Samaritans (phone: 116123), or Pieta House (1800 247 247).

More:
Meet the doctor giving free menopause support to LGBT people - RTE.ie

When researcher Arkasubhra Ghosh finally met Uditi Saraf, he hoped that there was still a chance to save her.

Ghosh and his collaborators were racing to design a one-off treatment that would edit the DNA in the 20-year-old womans brain cells and get them to stop producing toxic proteins. It was an approach that had never been tried before, with a long list of reasons for why it might not work.

CRISPR cures and cancer vaccines: researchers can help to shepherd them to market

But the team was making swift progress. The researchers were maybe six months away from being ready to give Uditi the therapy, Ghosh told her parents over breakfast at their home outside New Delhi last June. Even so, Uditis mother was not satisfied. Work faster, she urged him.

Then, Uditi was carried to the breakfast table, and Ghosh understood her urgency. Once a gregarious and energetic child and teenager, with a quick laugh and a mischievous streak, Uditi was now unable to walk or feed herself. She had become nearly blind and deaf. Her family tried to talk to her: These are the people who are making a therapy for you, they said loudly.

Shaken, Ghosh returned to his gene-therapy laboratory at Narayana Nethralaya Eye Hospital in Bengaluru, India, and got to work. If you need to put up tents in the lab, then we can do so, he told his students. Im not going to sleep.

Four months later, Uditi was gone.

The first therapy using CRISPR genome editing was approved in late 2023 to treat blood disorders that affect thousands of people worldwide. But the approach is also a source of hope to many people who have extremely rare genetic conditions, like the one Uditi had. Genome editing could one day become a radical way to address the diseases that are overlooked by pharmaceutical companies. Patients are waiting, families are waiting, says Jennifer Doudna, a molecular biologist at the University of California, Berkeley. So we need to get on with it.

How personalized cancer vaccines could keep tumours from coming back

Researchers are still laying the groundwork for this future. They are working out how best to design and manufacture the treatments, and how to deliver them to precise locations in the body. The cost is also a problem: the price of genome-editing therapy threatens to put it out of reach for many. Ghosh wants to bring those barriers down, and hes convinced that India will eventually be the country to do it.

But Uditis family could not wait the pace of scientific research was too slow. They needed a sprint, and a team of researchers willing to take on not only the scientific challenge, but also the emotional heft and high risk of failure involved in attempting something that had never been done. What we were trying to do was really almost in the realms of science fiction, Ghosh says.

And he remains convinced that, despite Uditis tragic death, the lessons learnt will help others on a similar path. It truly is a story of hope.

As a young girl, Uditi was always in a hurry. Seizing any excuse to celebrate whether it was a birthday or a festival she would buzz around the house getting ready hours ahead of everyone else, peppering her mother with urgent requests. She greeted family and friends with cuddles and kisses and brightened parties with her laughter and dancing.

For the first nine years, there was no hint of trouble. And when it began, it was just a flicker a few seconds here and there, when Uditi would zone out.

Shed switch back on again as if nothing had happened, and her mother, Sonam, wasnt sure if she should worry. But then Sonam saw nine-year-old Uditi drop a camera on the floor and become confused as to why it was no longer in her hand. A mothers hunch hardened: something was wrong.

Uditi Saraf with her mother, Sonam Saraf.Credit: Rajeev and Sonam Saraf

The physicians diagnosed her with epilepsy. When Uditis seizures became more pronounced and she began to struggle at school, Sonam and Rajeev, Uditis father, decided it must be something more. In 2017, they had part of Uditis genome sequenced.

It was a deviation from the standard treatment path, but the Sarafs were technologically savvy and financially well off. In India, as in many places in the world, genome sequencing was still uncommon, its roll-out slowed both by the costs and by the dearth of genetic information from people of Indian descent in genetic databases. Without such data, it can be difficult to interpret sequencing results.

Uditis results, however, were unambiguous: a single-base change in the gene that codes for a protein called neuroserpin caused tangled polymers to form in her brain cells, interfering with their function. Uditis neurons were dying.

This condition is called FENIB (familial encephalopathy with neuroserpin inclusion bodies), and the symptoms which can be similar to dementia usually manifest late in life. Elena Miranda, a cell biologist at the Sapienza University of Rome, runs the worlds only lab that focuses on the disease. She says that its possible that many cases of FENIB go unreported because physicians do not often sequence the genomes of older adults with dementia.

How CRISPR gene editing could help treat Alzheimers

But the most severe forms of FENIB strike early and are exceedingly rare. Miranda has known of only three other people with the same mutation that Uditi had. This form of the disease is very aggressive, she says.

Uditi and her parents embarked on a lonely journey familiar to many people with rare diseases. They had never heard of FENIB, and neither had Uditis physicians. Sonam did some research but couldnt bring herself to fully absorb what she found. We thought its not possible, she says. It cannot happen with our daughter.

The Sarafs studied what they could find online and tried the interventions available to them: Indian ayurvedic treatments, a ketogenic diet, special schools, seeing a slew of physicians and trying out various medicines. We shopped for doctors. We shopped for gods, says Sonam, but Uditis condition slowly worsened.

The three moved to upstate New York in 2018 to send Uditi to a school for people with disabilities. Her seizures intensified, and frequent muscle spasms made it hard for her to walk or drink from a glass. Her bright personality was dimming. The Sarafs discussed experimental treatments with Uditis new physician, epilepsy specialist Orrin Devinsky at NYU Langone Health in New York City. Devinsky mentioned a couple of options, one of which was CRISPR genome editing. Rajeev seized on the idea.

Uditis disease was caused by a mutation that converts a single DNA base from a G to an A. A variation on CRISPR genome editing, called base editing, could theoretically correct exactly this kind of mutation (see Precision gene repair).

Devinsky also emphasized the difficulties. At that time, base editing which was first reported in 2016 had never been tested in a clinical trial. The technique requires shuttling a relatively large protein and a snippet of RNA into affected cells. Researchers were struggling with how to perfect this delivery for many organs the brain being one of the most daunting.

Even if each of these hurdles were surmounted, at best, base editing might stop the production of neuroserpin clumps in some of Uditis neurons. The treatment was unlikely to reach all affected cells, and it was unlikely to clear the clumps that were already present or to regenerate neurons that had been lost.

But Rajeev and Sonam saw an opportunity for hope: perhaps such a therapy could slow down the progression of Uditis disease, buying time for scientists to develop another treatment that could repair the damage that had been done. The Sarafs were on board.

Devinsky assembled a team at NYU Langone Health with expertise in genome editing and neuroscience to conduct preliminary studies of the approach. The researchers pulled together what funding they could from other grants, and the Sarafs funded the rest. We will sell our house if we have to, Sonam said.

The pressure in the lab was intense, says team member Jayeeta Basu, a neuroscientist at NYU Langone Health. The team genetically engineered Uditis FENIB mutation into cells grown in the lab. When the cells initially didnt seem to behave as expected, Basu asked her graduate student to repeat the experiment five times. I was always pushing, she says. We had to be fast, but we also had to be diligent. There was no short cut.

Rajeev Saraf with his daughter Uditi.Credit: Rajeev and Sonam Saraf

In December 2019, the Sarafs moved back to India. Maintaining a home in the United States was expensive, and Uditi missed her extended family. Then the COVID-19 pandemic struck, and in January 2021, Uditi was hospitalized with severe COVID-19. She spent 20 days in the hospital and her health was never the same, says Sonam. Communication became increasingly difficult for Uditi and she began to pace the house incessantly, rarely even going to sleep.

The Sarafs decided to speed up the base-editing project by funding a second team in India.

Meanwhile, Devinsky had petitioned a US foundation to devise a different experimental treatment called antisense therapy for Uditi. The family flew from India to the United States twice for injections into her spine. The trips became traumatic as her ability to understand the world around her declined.

CRISPR 2.0: a new wave of gene editors heads for clinical trials

The treatments didnt work. And the experience taught Rajeev and Sonam how long it could take to get approval to try an experimental therapy in the United States. They decided Uditis base-editing therapy should also be manufactured and administered in India.

About an hour and a half away from their home, Debojyoti Chakraborty, a geneticist at the Council of Scientific and Industrial Researchs Institute of Genomics and Integrative Biology in New Delhi, had been making headlines for his efforts to devise a CRISPR-based treatment for a genetic blood disorder called sickle-cell disease.

Researchers in the United States were also developing genome-editing therapies for sickle-cell disease, but those therapies were expected to be expensive and potentially out of reach for much of the world. (The UK Medicines and Healthcare Products Regulatory Agency approved the first one, Casgevy, made by Vertex Pharmaceuticals in Boston, Massachusetts, and CRISPR Therapeutics in Zug, Switzerland, which costs US$2.2 million per patient.)

Most of the people with sickle-cell disease in India a country with one of the highest rates of the condition live in impoverished communities. Chakraborty and his colleagues hoped to develop a therapy that could be produced and administered at a fraction of the price that is charged in the United States, if not less.

Debojyoti Chakraborty is trying to develop affordable CRISPR-based treatments in India.Credit: RNA Biology Lab

Rajeev and Sonam went to the institute to talk to Chakraborty and the institutes director, chemist Souvik Maiti, who had been collaborating with Chakraborty on the CRISPR technology behind the sickle-cell project.

Move over, CRISPR: RNA-editing therapies pick up steam

Although the institute gets many requests for help from people with rare diseases and their caregivers, the Sarafs were unusual in that they would be able to help fund the work, says Maiti. Uditi was the only person in India known to have her neuroserpin mutation, and no government agency, company or philanthropy was likely to pay for the development of a treatment. Its very difficult, Maiti says. Even if our heart is telling us we should work on it, until there is funding, we cannot do it.

Even with funding, Maiti and Chakraborty took some time to discuss the project with Ghosh, who was building a facility in Bengaluru to produce viruses called adeno-associated viruses (AAVs), which are often used in gene therapies. Ghosh aimed for his facility to be one of the first in India to produce AAVs to the standards required for use in people.

There were a lot of unknowns in the base-editing project. And in addition to the work on stem cells in the lab, the team would need to do further experiments to determine which base-editing systems would work best, where and how to deliver its components into the body, and whether the process generated any unwanted changes to the DNA sequence. They would need to do experiments in mice to test the safety and efficacy of the treatment. They also needed to get Ghoshs facility approved by Indias regulators for producing the base-editing components.

Uditis illness had probably already progressed beyond the point at which the therapy could offer a notable benefit, but the family wanted them to try, reasoning that the work that they did on this project could help future endeavours to develop genome-editing therapies for genetic conditions.

It was not the first time Ghosh was swayed by a personal appeal: a few years before he met Uditi, Ghosh came to work and found two women waiting outside of his office. They would not leave, the women said, until he committed to finding a treatment for their young sons illness, a genetic condition called Duchenne muscular dystrophy, which can be fatal. The women pledged to help raise funds, and Ghosh found himself unable to say no. He has worked on the project and grown close to the families since then.

UK first to approve CRISPR treatment for diseases: what you need to know

Lab protocols for making medicines are notoriously strict, with each step carefully controlled to minimize the chance of contamination. When setting up his facility for manufacturing gene therapies, Ghosh scrutinized each step, looking for ways to cut costs without sacrificing safety, arguing his case to Indias regulators. He estimates that gene therapies for eye diseases that are developed in his lab will one day be available for one-hundredth of what they cost in the United States. We will certainly short circuit this entire field, Ghosh says.

India has earned a reputation for making complex drugs on a budget. During the COVID-19 pandemic, Indian manufacturers cranked out millions of doses of vaccines. Now, the country is manufacturing a malaria vaccine at a fraction of the cost of that in Europe, and it is developing sophisticated cell and gene therapies used in cancer treatments for much less than the price of those in the United States.

Chakraborty took the lead on Uditis project. He is a go-getter kind of person, says Riya Rauthan, who was then a PhD student in Chakrabortys lab. He is not bothered by who he needs to ask to get something done, he just does it.

To minimize interruptions, the team mapped out all of the experiments and the components they would need from start to finish. In India, many lab reagents have to be imported, and supply interruptions can delay projects by weeks or months. Everything had to be planned and ordered ahead of time, and Maiti worked to keep the supplies coming, seeking out vendors and negotiating prices. Time was more valuable than anything else, he says.

One of the most important reagents had to come from abroad: antibodies that could recognize the neuroserpin protein and its tangles. Few researchers use such antibodies, and the supply was uncertain. The team decided that the quickest way to get reliable antibodies was to ask Miranda in Rome to share the ones she had developed. She gladly did. This was a desperate approach, she says. But for me the priority was to try to help as much as I could.

Rauthan generated stem cells from samples of Uditis blood. Then, she and her colleagues coaxed those cells to become neurons, and used base editing on them in the lab.

Arkasubhra Ghosh is building an Indian facility to make viral vectors for gene therapy.Credit: Arkasubhra Ghosh

Ghosh worked on preparing the AAV that would be used to transport the CRISPR components into Uditis neurons. The team needed to determine which strain of AAV would work best some strains could trigger inflammation in the brain. Ghoshs lab tested several types of AAV in mice, to find out which one caused the least amount of inflammation and how best to administer it. The team eventually settled on one type called AAV9 and determined that it should be injected directly into Uditis brain.

Still, that was not the end of their challenges. AAV genomes can carry only an extra 4,700 DNA bases, but the gene that codes for the enzyme needed in base editing is longer than that. Ghosh and his students worked to divide up their genomic cargo so that it could fit in two separate viruses, and added sequences that would allow the two pieces to be spliced together again when they are expressed inside a cell. The team would inject both viruses at the same time.

The approach has been shown to work in mice but had not yet been tested in humans (J. M. Levy et al. Nature Biomed. Eng. 4, 97110; 2020).

By June 2023, the team seemed to be barrelling towards the finish line. Many of the researchers were working 10-to-12-hour days, and it was nearly time to test their therapy in mice. Ghosh was also scheduling a regulatory inspection to ensure that he would have the approvals he needed by the time the animal results were in. A surgeon had agreed to perform Uditis injection.

If all went well, they might be ready to treat Uditi in as little as six months, Chakraborty predicted.

In early October, a few months after Chakraborty and Ghosh had breakfast with Uditi and her parents, the team received a series of messages from Rajeev on their WhatsApp group. Uditi had become ill with pneumonia and had been taken to the hospital. Then she was in a coma and had been sent home there was nothing else the physicians could do for her.

Soon afterwards came the message they had all feared: Uditi was gone.

Ghosh thought immediately of the two little boys with Duchenne muscular dystrophy: What if Im too late for them, too?

Others in the lab also took the news hard. For clinicians, perhaps they become hardened, says Chakraborty. We dont have that experience. We were feeling agony.

Ten days after learning that Uditi died, Chakraborty presented the labs efforts at a local conference and finished his talk with a picture of Uditi, smiling. In the audience, Riya Patra, a graduate student in Ghoshs lab, began to cry. It was the first time shed let herself see a picture of the young woman shed tried so hard to save. Before, I had thought that if I saw her, maybe I would cry, she says. And I wouldnt be able to work anymore.

Is CRISPR safe? Genome editing gets its first FDA scrutiny

An estimated 100 million people in India have a rare disease. For decades, people affected by such conditions have cycled through hope and disappointment as researchers have inched closer to developing therapies that can help them at a genetic level. After a series of sporadic starts and failures, gene therapy has finally begun to find its footing. This has set the stage for CRISPR-based genome editing to rocket to the clinic.

When nine-year-old Uditi first dropped her camera, CRISPR was just an oddity a strange assembly of sequences found in microbial genomes, only studied by a few die-hard microbiologists. Four years before she was diagnosed with FENIB, researchers showed for the first time that a CRISPR-based system could cut DNA in human cells grown in the lab. And the first CRISPR-based genome-editing therapy was approved in the United Kingdom to treat sickle-cell disease the month after Uditi died.

In theory, many people with a genetic condition, no matter how rare, could benefit from these technologies. But the reality is harsher. It will take years to establish the techniques needed to create rapid, on-demand, bespoke CRISPR therapies. Most people with these conditions dont have that kind of time.

But researchers are working to streamline the process. Doudnas institute, for example, is working to standardize some aspects of genome-editing therapies, in part to make it cheaper and easier to develop such treatments for people with rare conditions. And the US National Institutes of Health has been trying to develop similar pipelines for gene therapies an effort that could help to inform genome-editing efforts. Its been really hard, says Doudna. But what were doing is going to have long-term impact.

In India, the work has continued. Rajeev has urged Chakraborty to finish the teams studies in mice, so that the next person with FENIB will not have to wait as long for a potential treatment. Some of the work will be completed, and the effort could benefit others with genetic conditions that affect the brain particularly in India. We are not really trying as aggressively as we did earlier, he says. But that technology has a lot of potential.

At Uditis memorial service, Rajeev tried to make sense of the timing. Uditi was always in a hurry, he told attendees. She always had to be first. She was only a few months away from receiving an experimental treatment, but she would not wait, not even for that. She could not let science win, he said. She was always ahead.

Link:
Hope, despair and CRISPR the race to save one woman's life - Nature.com

More than 30,000 people in the United States alone have already received personalized CART-T-cell therapy for cancer.Credit: Qilai Shen/Bloomberg/Getty

When researchers first began to test engineered immune cells designed to fight cancer about 20 years ago, there was a scepticism. The scientific potential might be clear, but what about the economics of such a complex and specialized therapy? Each dose would have to be made afresh, with cells from an individual being shipped to a centralized laboratory, genetically engineered using sophisticated techniques and shipped back for reinfusion. The process would take too long and be too expensive. Regulators would also surely struggle to ensure the safety of such an involved, individualized process.

Today, the chatter is very different. Engineered CAR T immune cells have so far been used to treat more than 30,000 people with cancer in the United States alone. CAR-T therapy is being tested for other conditions, including some severe autoimmune disorders. As for commercial success, in 2023, CAR T cells earned biotechnology companies US$8.4 billion worldwide.

Two News Features in this issue describe other complex, bespoke therapies that, a decade ago, would have been considered infeasible, if not impossible. One is an mRNA cancer vaccine tailored to an individuals tumour genome. The other is a CRISPR-based genome-editing therapy designed but sadly never used for one young woman with a rare neurological disorder.

How personalized cancer vaccines could keep tumours from coming back

Both approaches are fraught with challenges. As in the early days of CAR-T therapy, many of them are not scientific. But by guiding regulators and developing flexible platforms for producing bespoke treatments, researchers can help to shepherd therapies to the people who need them.

Researchers have long chased after vaccines that could rally the immune system against tumours, similarly to how vaccines rouse defences against pathogens. Companies can now sequence portions of a persons tumour and select those most likely to be visible to the immune system. The mRNA molecules corresponding to those regions are synthesized, then encapsulated in fatty particles and injected much like mRNA COVID-19 vaccines. From start to finish, the process takes as little as a month.

The technology behind these cancer vaccines is clinically more advanced than the genome editing used for some more specialized applications, for which researchers do not have the luxury of running large clinical trials. In one instance, scientists knew of only one person with the mutation they aimed to treat, using a technique called base editing that can make changes to specific DNA bases. It was, in effect, a treatment designed for a market of one person.

This kind of approach is called an n-of-1 therapy, a term that highlights the statistical challenges of interpreting results from a sample of one not to mention the commercial challenge of designing and selling a therapy with a one-person market. But the name is potentially misleading and stigmatizing. A cancer vaccine based on an individuals tumour could also be considered an n-of-1 therapy, yet this approach has attracted heavy investment from the pharmaceutical industry because the same process can be extended to many other people with cancer.

Hope, despair and CRISPR the race to save one womans life

The same thinking is needed for genome-editing therapies for rare disorders. Some genetic conditions that weaken or disable the immune system could be grouped together, and therapies for these diseases designed and administered in the same manner, even if the specific DNA changes made are different. Identical or similar measures such as levels of immune-cell function could be used to determine how well the treatment works.

But for-profit companies cannot be relied on to develop such platforms for CRISPR-based therapies as long as the perceived market remains small. Some academic researchers are focusing on developing such platforms for CRISPR-based therapies. More should join them or the chance to use genome editing to correct genetic disorders, the most severe of which are often rare, will be squandered.

Researchers can help regulatory agencies grappling with the new technologies. Regulators in the United States, the European Union, India and the United Kingdom have signalled a wish to aid the development of treatments for ultra-rare disorders. But they need help. Many regulations governing the manufacturing of therapies are grounded in regulatory paths forged years ago. Scientists can advise regulators on which technological advances have rendered certain cumbersome regulations unnecessary. This could speed up the development of treatments, as well as lower their costs.

Researchers around the world can engage in the same discussions with their regulators, and not just in typical hotspots for drug development, such as the United States and Europe. Such conversations will help to prepare for a future in which bespoke genetic therapies can be produced worldwide. They could also help to harmonize regulations between countries: an important goal for promoting the development of drugs for conditions that affect only a few individuals scattered around the globe.

As data accumulate from the treatment of people with rare genetic disorders, lessons learnt about the safety, effectiveness and manufacturing of bespoke therapies can be translated to treatments for more-common conditions. So the treatment of ultra-rare genetic disorders should not be devalued. Although a single disorder might affect only a few people, in aggregate, ultra-rare diseases affect millions. When it comes to personalized medicine, serving the interests of the few is in the interests of the many.

Continued here:
CRISPR cures and cancer vaccines: researchers can help to shepherd them to market - Nature.com

Over the past decade, CRISPR has taken the biomedical world and life sciences by storm for its ability to easily and precisely edit DNA. Here, Stanford University bioengineer Stanley Qi explains how CRISPR works, why its such an important tool, and how it could be used in the future including current developments in using CRISPR to edit the epigenome, which involves altering the chemistry of DNA instead of the DNA sequence itself.

CRISPR is not merely a tool for research. Its becoming a discipline, a driving force, and a promise that solves long-standing challenges from basic science, engineering, medicine, and the environment, said Qi, an associate professor in the Department of Bioengineering and institute scholar at Sarafan ChEM-H. Together, we can think innovatively about how to match needs with technologies to solve the most challenging problems.

(click the question to jump to the answer):

What is CRISPR

How does it work?

What are gene therapy and cell therapy, and how is CRISPR involved?

How does it differ from other gene-editing tools?

Why is it such a big deal?

How far has CRISPR technology come since it was created?

In 2019, Victoria Gray was the first person in the U.S. to receive CRISPR treatment for a genetic disease (sickle cell anemia). Now, CRISPR-based therapies are approved in the U.S. and the U.K. What is next?

Were you surprised when the 2020 Nobel Prize in chemistry went to CRISPRs developers?

Besides treatment for diseases, what are other real-world applications for CRISPR technology?

What are your views on some of the ethical concerns surrounding CRISPR?

Your group demonstrated that its possible to shrink CRISPR. Why is this significant?

What is your lab working on in terms of epigenome editing?

Are there limitations to what CRISPR can do?

What do you think CRISPR is capable of doing in the future?

How far are we from actually achieving those idealistic future goals?

The short answer: CRISPR is an immune system used by microbes to find and eliminate unwanted invaders.

Qi: CRISPR stands for clustered interspaced short palindromic repeats. Biologists use the term to describe the genetic appearance of a system that was discovered in microbes including bacteria and archaea as early as 1987. For a long time, no one really understood what it did, but around 2005, researchers discovered CRISPR is an immune system. Its used by microbes to help protect themselves from invading viruses. To stop the invaders, the microbes use CRISPR to recognize and eliminate specific trespassers.

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The short answer: When a virus or other invader enters a bacterial cell, the bacterium incorporates some of the trespassers DNA into its own genome so it can find and eliminate the virus during future infections.

Qi: Its similar to the human immune system. When a virus infects us, we generate an immune memory in the form of antibodies lots of them. Then, when the same virus infects us again, these antibodies quickly recognize the invaders and eliminate them.

When a virus infects a bacterial cell, CRISPR helps establish a memory a genetic one. The bacterium takes a piece of the viruss genome and inserts the DNA into its own genome. From that newly acquired DNA sequence, CRISPR creates a new guide RNA, a sequence that helps CRISPR find the invader via sequence complementarity (i.e., A binds to T and C binds to G). So, the next time when the virus infects that bacteria cell, the guide RNA rapidly recognizes the virus DNA sequence, binds to it, and destroys it.

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The short answer: Gene therapy can mean using CRISPR as a macromolecule drug to either fix a mutated gene or regulate a defective gene to treat a disease. Cell therapy means using CRISPR to make your bodys cells attack toxic cells or regenerate beneficial cells.

Qi: Gene therapy can mean two things: One is to fix a mutated gene, and the other is to regulate a genes expression into protein products. Our current understanding of gene therapy is still rapidly advancing, and the challenge is managing therapy safely and cheaply. Furthermore, were only looking at the simplest genetic diseases. For example, sickle cell anemia is a disease we know a lot about, and its often caused by a single mutation. So, we can configure CRISPR to fix it. But many more diseases are caused by widespread mutations, multiple mutations, and even multiple genes. In the future, gene therapy could go beyond a single mutation, and I am optimistic that in the next decades, gene therapy will become a pillar of medicine.

Cell therapy is a little different. For example, when people try to treat leukemia, a type of white blood cell tumor, sometimes chemotherapy drugs cant completely get rid of the tumor cells. In the past two decades, scientists have found that if they retrieve some of the patients T cells, which fight infections, these cells can be engineered as better fighters to recognize and eliminate tumorous cells. When the modified T cells are injected back into the patient, they can attack the tumors. However, cells are quite complicated. Sometimes, they go out of control when injected back into the patient, killing healthy cells along with the tumor cells. At other times, they may fail to work because they are suppressed by the tumor cells. CRISPR offers a powerful tool to enhance the efficacy and safety of these immune cells so that they are completely under our control for best clinical benefits.

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The short answer: CRISPR is much easier to program than other tools.

Qi: Before CRISPR, most gene-editing tools were a single protein. By changing the peptide sequence of these proteins, scientists could alter their targets. To change the target, you need to completely redesign the proteins sequence and then test if it even works, which is tedious, unpredictable, and time-consuming. These gene-editing tools were theoretically interesting, but they were difficult to use for large-scale studies and therapeutics.

Compared to that, CRISPR is elegant because the target recognition sequence is mostly encoded within an RNA rather than a protein, and redesigning this sequence is one of the simplest things you can do in molecular biology. It makes genome editing similar to operating a GPS: If you want to go to destination A, you just type the address, and to change to destination B, you just enter the new location. So, this tool dramatically reduces the burdens, cost, timing, while increasing the precision and accuracy of a gene-editing system.

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The short answer: CRISPR can precisely modify a piece of DNA or its chemistry (so-called epigenetics) in the human body, making it a potential tool for clinical uses in the biomedical sciences.

Qi: CRISPR is a molecule and tool desired by everyone who works in the life sciences, biomedical research, and clinical settings. Its high precision is unparalleled and enables many uses including gene therapy.

My dream has been to develop new biotechnologies and apply them to diseases without a cure. Genetic diseases make up a big part of this category. Traditional medicines small molecule drugs, surgery, and other methods dont work for these types of diseases. But CRISPR molecules have become highly promising as treatments because they allow us to precisely modify a piece of DNA in the human body. This could lead not only to relief but also to a cure.

Indeed, recent FDA approval of the first CRISPR drug, Casgevy, in treating sickle cell anemia and beta thalassemia speaks to its safety and potential for other diseases. Sickle cell anemia is a disease in which people have a mutation in their red blood cells. Normally, theres no treatment other than frequent blood transfusions or bone marrow transplants from a matched donor, which are expensive and damaging to a patients overall health. Using CRISPR, its possible to perform a one-time treatment to permanently correct the mutation. There are more than 8,000 genetic diseases like that, which can be potentially considered.

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The short answer: In about a decade, scientists went from wondering if this technology would even work in human cells to getting the first CRISPR drug approved uses in the clinic.

Qi: In 2010, I was working on CRISPR as a bioengineering graduate student at the University of California, Berkeley, under Adam Arkin, a synthetic biologist and bioengineer, and collaborated with Jennifer Doudna, a biochemist and structural biologist. In the early days, CRISPRs practical usefulness was not very publicly recognized. At that time, many counterarguments said CRISPR was just a bacterial system and most of these simply dont work in human cells which, to be fair, is true.

But after Jennifer Doudna and Emmanuelle Charpentier published their seminal 2012 paper on Cas9 one type of CRISPR that cuts DNA using a single protein and an engineered single guide RNA the research and published papers grew exponentially. Firstly, because its a system that everyone in the life sciences wants. Secondly, using CRISPR is super easy, flexible, and robust. Its not like other technologies that take multiple years and millions of dollars to set up CRISPR only takes a couple of weeks and a bit more than a few hundred dollars to set up now.

A lot of researchers significantly contributed to the rapid development. For example, within three years following its initial demonstration, structural biologists solved the high-resolution, three-dimensional structure of what Cas9 and other CRISPR proteins look like. Bioinformaticians have revealed many new species of Cas molecules beyond Cas9, many of which have novel functions. Biochemists engineered CRISPR to understand how fast and tightly it binds to DNA. Bioengineers, including me, engineered the proteins to make them work more efficiently and more specifically so they can work better in the human body for gene therapies. Also, clinical researchers started to use the tool to address particular diseases.

Furthermore, the applications of CRISPR went beyond gene editing. Epigenetic editing is an exciting development, although we still await clinical benefits. It was used for targeting the human 3-dimensional genome, visualizing the DNA dynamics, or even targeting another set of molecules, RNA, for gene regulation.

I dont think Im exaggerating to say that, essentially, CRISPR has been tested as a potential treatment option for every disease that we have clear knowledge about. CRISPR cant solve all of them, but because this tool is so powerful, easy to use, and so far-reaching, it has allowed everyone to combine their expertise with CRISPR.

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The short answer: This is very exciting. Future CRISPR drugs will address more incurable diseases, which provide a test case for CRISPRs efficacy and safety in different organs and patients.

Qi: Im super excited to see CRISPR becoming a drug to treat a disease as a one-time cure. When CRISPR first came out, there were concerns about whether these bacterial molecules could be used safely in humans and whether it was safe to cut and edit human DNA. While there are still questions regarding long-term effects (beyond the period of clinical trials in tested patients) it is very encouraging that CRISPR is safe and effective.

The next step is to expand the scope of CRISPR drugs. Medicine isn't made in one day. Different diseases are caused by different mechanisms. There are already more than dozens of CRISPR clinical trials for different diseases in the liver, immune cells, eyes, and muscles. Furthermore CRISPR epigenetic editing is expanding the scope of disease to treat more types of muscular dystrophy, retina disorders, and brain diseases.

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The short answer: Not at all. But I hope the award doesnt lead people to think CRISPR research is finished its still growing, and theres much more to explore in basic research, medicine, and beyond.

Qi: Im not surprised at all. Even before 2020, researchers had been discussing when the Nobel Committee would recognize CRISPR. So, when it happened, I was super excited.

Jennifer Doudna (University of California, Berkeley) and Emmanuelle Charpentier (Max Planck Unit for the Science of Pathogens) received the Nobel Prize in Chemistry only seven years after CRISPR was first reported as a molecular system for modifying the human genome.

I hope that giving the Nobel Prize to CRISPR wont give people the impression that the genome editing field is done. This is a field thats still growing in every corner of life sciences. Besides being explored as medicine in humans, it is expanding its influence in plants, microbes, and difficult-to-engineer organisms such as fungi. There are so many questions about how we can use CRISPR for safely controlling the genome, how to use it for novel and innovative research, and how to make it a clinical product that still need to be explored.

These are exciting frontiers of further increasing the safety of CRISPR-based therapies and expanding the scope of diseases treatable by this technology.

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The short answer: Some other uses are diagnostics, manufacturing, sustainability, and ecological engineering.

Qi: CRISPR can be used for diagnostics. It has been developed as a way to sensitively detect pathogens in the environment that are affecting our bodies.

There are also opportunities in manufacturing, such as making products that we care about using organisms like yeast and bacteria. Imagine that we could use CRISPR to engineer new microbes that could boost production like 10x more beer, for instance. And also, beer that tastes much better and can be catered to different peoples wants and needs.

Sustainability is also a big application for CRISPR via bioengineering. Creating sustainable, carbon-neutral methods of energy or food production is a challenge. Genome engineering may offer better manufacturing protocols through microbes that reduce greenhouse gases, plastic, and food waste.

Finally, we get to ecological engineering. For example, people are trying to eliminate certain invading or pathogenic mosquito species using CRISPR, but in my opinion, its long-term safety and impact still need careful evaluation. Other people are trying to revive extinct species. Recently, scientists announced they were trying to revive a woolly mammoth that can live in the Arctic cold.

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The short answer: My research group often thinks about the ethics of CRISPR. Some ethically questionable areas are disease prevention and eliminating pesky species, and some definite unethical areas are enhancement and creating designer babies.

Qi: The ethical side of CRISPR is something my research group thinks about every day. One of the fundamental principles of ethics is to do no harm. Sure, we want to do something great and helpful to people, but at the same time, we have to consider if were harming other people. Using that principle, we can consider a few cases.

One example is a designer baby, which is a scary topic. That is regarded as unethical because this may create a new human species. When the germ cells sperm and egg cells are edited, this not only affects that single person, but also the children that person could have in the future.

Another concern is in the division of treatment, which has three categories: cure, prevention, and enhancement. Curing someones disease is great. Prevention, which means someone is at risk of developing a problem, is a gray area. If someone has a high chance of getting an infectious disease, should we use gene therapy to permanently modify their DNA to reduce their risk? That question really depends on if we have other options. The last category enhancement is likely unethical. People talk about the possibility of targeting a gene to grow more muscle or make people smarter or better looking. But if research goes into this category, only some people may be able to afford it. This could amplify the imbalance of socioeconomic status. Another facet to consider is medical necessity. Is the therapy really necessary, or are there other ways to solve the problem through currently available drugs, diet, exercise, etc.?

Beyond medicine, some scientists may want to use CRISPR for ecological reasons, for example, eliminating mosquitoes. From my viewpoint, thats controversial because I think every species exists for a reason. If we try to eliminate mosquitoes, we might have a chain reaction that affects other life forms in the environment and can be irreversible. I hope in the future we can make this technology reversible like installing a switch so that if we make something that turns out to be less than ideal, we still have some way to reset it.

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The short answer: Its tricky to deliver CRISPR molecules into cells. Shrinking the size of the molecule helps it easily traverse inside of cells and get to its DNA target.

Qi: CRISPR is such a magic molecule, but that magic only works if CRISPR gets inside cells and touches the DNA. The question is obvious: How can we even make CRISPR get inside the cell?

Human cells are designed to resist any invading DNA. So the human body has many strategies to prevent foreign DNA from getting in.

Many delivery methods scientists used have limited power. We can use retooled viruses to deliver clinical products into cells, but they have a small capacity the Cas9 version of CRISPR usually doesnt fit inside the virus. Therefore, the currently approved CRISPR drug requires isolating patient cells, modifying them, and putting them back in. This process is costly and slow. If we want CRISPR to become a broadly useful medicine, then we need to make the molecule as small as possible.

Thats why we made this miniature CRISPR, which we call CasMINI, which is only half the size of Cas9. We also saw that it is easier to enter cells and works better than other CRISPR molecules because it can get inside more efficiently. This miniature CRISPR can revolutionize the way that we can perform editing in the body. Our hope is to address these technical barriers then test how miniature CRISPR can be delivered to different parts of the human body to treat various genetic diseases.

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The short answer: Were trying to use CRISPR to control gene function rather than editing genes to treat diseases.

Qi: Im excited about exploring how to treat diseases without modifying human DNA through epigenome editing. Its a different way of thinking about gene therapy. Unlike gene editing, epigenome editing is reversible, safer, and promising for complex diseases that can not be easily targeted by gene editing.

To enable epigenome editing, we developed the first nuclease-deactivated dCas9 in living cells, to programmably target and control gene expression, without altering the DNA sequence. For example, if a person doesnt have enough properly working proteins, we can use epigenome editing to increase the gene expression over a long term to make more proteins to compensate for this deficiency problem, thus restoring the function to normal in patients.

In other cases, someone may have a gene mutation that produces a toxic product, such as in many muscular dystrophies or neurological degenerative diseases. Rather than using CRISPR to modify DNA, we can use our epigenome editing technology to permanently silence the gene without modifying the DNA. I am excited to test this solution in the clinic as I believe this offers a safer strategy for treatment without altering DNA.

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The short answer: There are limitations to gene editing, but new technologies are trying to expand the power of CRISPR.

Qi: One major limitation is weve been using it for only 10 years. Often, time is the best test of all technologies. Only by collecting data over enough time in all scenarios will we be able to understand everything about these technologies, like how safe they are over the long term.

In testing in human subjects with patients, even though we didnt see off-target effects or immune responses, there are still question marks. We still need to constantly improve our understanding, as well as CRISPRs accuracy and precision in different human tissues and different patients, when treating a problem.

Also, right now, CRISPR is mostly used as molecular scissors to cut DNA. But sometimes, the problem genes affected function isnt caused by a DNA mutation. Sometimes, its a gene turning on or off abnormally that causes the problem. So in that case, CRISPR shouldnt be used as molecular scissors to cut DNA, but rather as a switch to restore the gene to work properly. Epigenetic editing tools can well address such challenges.

CRISPR is like a powerful hammer. But the question is: Where is the nail? What is the most suitable nail to work on? For example, as of today, we still dont know for sure which gene causes Alzheimers disease in many patients. To use CRISPR, we need to know which gene to target and which cell is the destination. We also need to know when to perform the treatment sometimes treatment can only be done in an early stage of a persons life.

Another big issue is the high costs associated with the current CRISPR medicine. How to reduce cost is a major question. Im glad that there are active conversations between academia and industrial partners to have multiple experts in the same room to come up with the best solution.

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The short answer: It could help improve the quality of life as we age, engineer useful organisms, and even serve as a universal vaccine against viruses.

Qi: Im excited by CRISPR possibly helping anti-aging, but less in the sense of making people live longer. No one can escape aging, and its a huge burden to our healthcare system and decreases the quality of life. My hope is that in the future, CRISPR isnt just being used to save lives, but also to improve the quality of life when people age.

I also hope CRISPR can become a way to engineer a lot of useful life forms. For example, there are microbes that can capture solar energy and convert it to electricity, and maybe those could be used to produce sustainable energy. Additionally, we could engineer food thats more nutritious, prevents obesity, and so on.

Another application could be vaccines. Even now, infectious diseases, like COVID-19, have dramatically changed everyones lives, which is unbelievable. So another dream is to develop cheap and safe genetic vaccines to fight all viruses, since thats their original role in bacteria. And maybe, in the future, we could receive a small dose of CRISPR that could completely kill any new virus. Its not easy, but given that this genetic system was designed as an antiviral system, theres a chance this could work.

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The short answer: Were close to some goals but may be far from some other idealistic goals.

Qi: When it comes to CRISPR and achieving those big dreams we have for it, we're at different stages. For some goals, it might feel like we're just starting out, but for others, we're getting pretty close. For example, I'm really excited about how we're starting to use CRISPR in real-life treatments for diseases, such as sickle cell anemia. This is a big step forward! I am also very excited about CRISPR epigenetic editing, a way to turn genes on or off without changing DNA sequence, which is getting ready for its big moment in clinical trials.

The reason weve come this far is thanks to a lot of people who believe in the power of safely editing our genes to make us healthier and are working hard every day to make that a reality. Its their passion and the demand for these solutions that keep pushing us forward. Im optimistic that many of the things were dreaming about with CRISPR could become real, sooner rather than later.

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What is CRISPR? A bioengineer explains | Stanford Report - Stanford University News

As some women age, their white blood cells can lose a copy of chromosome X. A new study sheds light on the potential causes and consequences of this phenomenon.

Credit: Created by Linda Wang with Biorender.com

Researchers have identified inherited genetic variants that may predict the loss of one copy of a womans two X chromosomes as she ages, a phenomenon known as mosaic loss of chromosome X, or mLOX. These genetic variants may play a role in promoting abnormal blood cells (that have only a single copy of chromosome X) to multiply, which may lead to several health conditions, including cancer. The study, co-led by researchers at the National Cancer Institute, part of the National Institutes of Health, was published June 12, 2024, in Nature.

To better understand the causes and effects of mLOX, researchers analyzed circulating white blood cells from nearly 900,000 women across eight biobanks, of whom 12% had the condition. The researchers identified 56 common genetic variantslocated near genes associated with autoimmune diseases and cancer susceptibilitythat influenced whether mLOX developed. In addition, rare variants in a gene known as FBXO10 were associated with a doubling in the risk of mLOX.

In women with mLOX, the investigators also identified a set of inherited genetic variants on the X chromosome that were more frequently observed on the retained X chromosome than on the one that was lost. These variants could one day be used to predict which copy of the X chromosome is retained when mLOX occurs. This is important because the copy of the X chromosome with these variants may have a growth advantage that could elevate the womans risk for blood cancer.

The researchers also looked for associations of mLOX with more than 1,200 diseases and confirmed previous findings of an association with increased risk of leukemia and susceptibility to infections that cause pneumonia.

The scientists suggest that future research should focus on how mLOX interacts with other types of genetic variation and age-related changes to potentially alter disease risk.

Mitchell Machiela, Sc.D., M.P.H., Division of Cancer Epidemiology and Genetics, National Cancer Institute

Population analyses of mosaic X chromosome loss identify genetic drivers and widespread signatures of cellular selection appears June 12, 2024, in Nature.

About the National Cancer Institute (NCI):NCIleads the National Cancer Program and NIHs efforts to dramatically reduce the prevalence of cancer and improve the lives of people with cancer. NCI supports a wide range of cancer research and training extramurally through grants and contracts. NCIs intramural research program conducts innovative, transdisciplinary basic, translational, clinical, and epidemiological research on the causes of cancer, avenues for prevention, risk prediction, early detection, and treatment, including research at the NIH Clinical Centerthe worlds largest research hospital. Learn more about the intramural research done in NCIs Division of Cancer Epidemiology and Genetics. For more information about cancer, please visit the NCI website atcancer.govor call NCIs contact center at 1-800-4-CANCER (1-800-422-6237).

About the National Institutes of Health (NIH):NIH, the nation's medical research agency, includes 27 Institutes and Centers and is a component of the U.S. Department of Health and Human Services. NIH is the primary federal agency conducting and supporting basic, clinical, and translational medical research, and is investigating the causes, treatments, and cures for both common and rare diseases. For more information about NIH and its programs, visitnih.gov.

See the original post:
New study offers clues into genetics of X chromosome loss - National Cancer Institute (.gov)

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Genetic drivers and cellular selection of female mosaic X chromosome loss - Nature.com

Richard DiGeorge and Jo Kaur, pictured with their son Riaan DiGeorge, founded the nonprofit Riaan Research Initiative to support UMass Chan scientists quest to cure Cockayne syndrome

UMass Chan Medical School has received $2.2 million from a nonprofit patient-advocacy organization to contract with Ohio-based Andelyn Biosciences, a contract development and manufacturing organization with extensive experience in gene therapy, to manufacture clinical grade AAV9-CSA vector to treat Cockayne syndrome.

The gift from Riaan Research Initiative brings the start of clinical trials for the fatal autosomal recessive disorder one step closer.

Riaan Research Initiatives historic contribution is the largest known donation ever made toward the development of a treatment for Cockayne syndrome, according to Riaan Research Initiative.

Cockayne syndrome primarily causes mutations in genes CSA (ERCC8) and CSB (ERCC6) and leads to a failure in DNA transcription and repair processes. Impacted children often present with significant growth failure, microcephaly, vision and hearing problems, and global developmental delays. Children with the most severe form of Cockayne syndrome have a life expectancy of five to seven years. There are no approved treatments.

Were thrilled to move on to the next step of this years-long partnership with UMass Chan and are eternally grateful to our donors who have opened the doors to a better world, said Jo Kaur, founder of Riaan Research Initiative and mother of Riaan, a 4-year-old boy diagnosed with Cockayne syndrome and the inspiration behind the research. We have strong evidence of the treatments success in the preclinical phase and look forward to our collaborators manufacturing a potentially life-saving drug that can actually be given to our children.

UMass Chan and Riaan Research Initiative launched this collaboration in 2021 when the organization funded the preclinical phase of this groundbreaking program. Now that research led by Miguel Sena-Esteves, PhD, associate professor of neurology and director of the Translational Institute for Molecular Therapeutics, and Rita Batista, PhD, instructor in neurology, has demonstrated the AAV9-CSA vector efficacy in an animal model, the team is ready to move forward with toxicology studies and clinical manufacturing.

Andelyn Biosciences has been selected to manufacture the plasmids and use its AAV Curator Suspension Platform to industrialize the process, performing analytical development and process optimization, followed by toxicology manufacturing and GMP (good manufacturing practice) production.

Earlier this year, the UMass Chan research team and Riaan Research Initiative received favorable feedback from the U.S. Food and Drug Administration on their pre-Investigational New Drug submission with plans for toxicology studies and manufacturing.

Riaan Research Initiative has been an amazing partner in our journey to develop an AAV9 gene therapy for Cockayne syndrome, Dr. Sena-Esteves said. Our dream of making a difference for Cockayne patients is coming closer every day, and signing the contract with Andelyn Biosciences to make the clinical material is a major step in that direction. Developing a gene therapy for fatal pediatric neurological diseases is challenging in many ways, but together with Riaan Research Initiative we have an outstanding team to bring our program to a first-in-human gene therapy clinical trial for Cockayne syndrome.

Original post:
UMass Chan receives $2.2 million to fund gene therapy for Cockayne syndrome - UMass Medical School

The risk of secondary cancer after CAR-T therapy, pioneered at Penn, is less than regulators feared last year, an FDA official said Friday at cell and gene therapy conference in King of Prussia.

The FDA had announced in November that it was studying a handful of cases where patients developed lymphoma after being treated with chimeric antigen receptor T cell, CAR-T, therapies, hailed as a possible cure for some forms of blood cancer.

In January, the agency ordered the companies to put black box warnings on the six products that were part of the review.

We were pretty concerned about this when we first saw it last year, Peter Marks, who heads the FDAs Center for Biologics Evaluation and Research, said at the conference sponsored by the Sino-American Pharmaceutical Professionals Association Greater Philadelphia.

Most look to be secondary cancers affecting the T cells of patients who already have related cancers, he said.

Luckily, the majority of these, it looks like are just secondary T cell malignancies that are occurring in people who have T cell malignancies. Thats a known phenomenon, he said.

In a few cases, however, there are signs that patients developed secondary cancer in the type of white blood cells that were genetically modified as part of their treatment, Marks said. The risk is probably on the order of 1 in 10,000 people treated, he said.

That risk is orders of magnitude lower than the risk of malignancies from forms of chemotherapy that are given after cancer hasnt responded to standard treatments, Marks said during his opening keynote address at the conference, now in its third year.

About 500 people were registered for the 2024 @Philly Cell And Gene Therapy Annual Conference, a spokesperson for the conference organizers said.

READ MORE: Emily Whitehead was the first child cured of cancer with therapy from Penn. Shes back as a freshman.

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Philadelphia Cell and Gene Therapy Conference hosts FDA officials - The Philadelphia Inquirer

A Pfizer gene therapy for the rare muscle-wasting disease Duchenne muscular dystrophy did not help patients ability to walk or stand up in a pivotal clinical trial. The pharmaceutical giant is still weighing its next steps for the therapy, but analysts say these disappointing trial results, the latest in a series of setbacks, likely mark the end for this program.

The Phase 3 test enrolled boys ages 4 to 7 who still had the ability to walk. The trials main goal was to show an improvement in motor function compared to a placebo. Without providing specific figures, Pfizer said after Wednesdays market close that its gene therapy did not achieve this goal measured one year after treatment. Secondary goals include measuring how fast patients can run or walk for 10 meters and how quickly they can rise from the floor. On these measures, there was no significant difference between the gene therapy arm and the placebo group.

We are extremely disappointed that these results did not demonstrate the relative improvement in motor function that we had hoped, Dan Levy, Pfizers development head for Duchenne muscular dystrophy, said in a prepared statement. We plan to share more detailed results from the study at upcoming medical and patient advocacy meetings, with the goal of ensuring that learnings from this trial can help improve future clinical research and development of treatment options that can improve care for boys living with Duchenne muscular dystrophy.

Duchenne is an inherited disorder that results in the inability to produce normal versions of dystrophin, a protein key to muscle function. Patients develop progressively worsening muscle weakness that robs them of their ability to walk. The muscle weakness eventually affects the lungs and the heart, becoming fatal.

[Paragraph updated to correct the study on dosing pause.] The Pfizer gene therapy, fordadistrogene movaparvovec, uses an engineered virus to deliver to muscle cells a mini-version of the gene that codes for dystrophin. Its clinical development path has had prior setbacks. A patient death in 2021 led to a clinical hold on tests of the therapy. Pfizer was later cleared to resume clinical trials after implementing additional safety measures. But last month, Pfizer disclosed a patient death in a Phase 2 study evaluating its Duchenne gene therapy in boys ages 2 to 3. Dosing in that study is complete. Pfizer said dosing in the crossover arm of the Phase 3 study has paused as the company continues to gather information to understand what caused the patient death in the Phase 2 test. In the latest results reported Wednesday, Pfizer said the gene therapys safety profile was manageable and adverse effects were mostly mild to moderate.

The developments unfolding for Pfizers gene therapy come as the FDA weighs whether to award full approval to Elevidys, a Duchenne gene therapy developed by Sarepta Therapeutics. Nearly a year ago, the Sarepta therapy won accelerated approval for Duchenne patients ages 4 and 5. But last fall, the company reported the failure of the Phase 3 study meant to confirm the therapys benefit and support expansion to a wider range of patients. That pivotal study evaluated patients with the same measures used for Pfizers pivotal study.

Leerink Partners analyst Joseph Schwartz draws distinctions between the two gene therapy programs. Though both failed in Phase 3, Sareptas therapy showed statistically significant improvement according to its trials secondary measures, which are more sensitive in detecting benefit, he said in a Thursday research note. Furthermore, Sareptas gene therapy does not have the safety questions overhanging the Pfizer gene therapy.

Thus, with no efficacy signals and a less-than-pristine safety profile, we see this readout as the final nail in the coffin for the program and think it is unlikely to move forward, Schwartz said.

William Blair analyst Tim Lugo said in a research note that his firm did not view Pfizers gene therapy as a real competitive threat to Sarepta due to the safety concerns throughout its development. He echoed Schwartzs comments about the Sarepta therapys ability to hit the secondary goals of its study.

We believe overall the totality of the data generated to date support the efficacy of Elevidys, the current 4- and 5-year-old label, a conversion to full approval, and a broader expansion to include older boys and into non-ambulatory patients, Lugo said. However, we believe expansion into non-ambulatory patients is more of a stretch, and we would not be surprised if these patients are excluded from the expanded label.

Sareptas Elevidys faces a June 21 target date for an FDA decision.

Photo: Dominick Reuter/AFP, via Getty Images

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Pfizer & Sarepta Gene Therapies Both Failed Phase 3, But Analysts Expect Sarepta Will Win Approval - MedCity News

Expanding on his earlier podcast discussion with EPR, Dr Arun Upadhyay, Chief Scientific Officer and Head of Research & Development at Ocugen, discusses the companys promising modifier gene therapy candidates for ophthalmic disorders.

OCU400 is a modifier gene therapy aimed at treating retinitis pigmentosa and Leber congenital amaurosis (LCA).

In April 2024, Ocugen received US Food and Drug Administration (FDA) clearance to initiate the Phase III liMeliGhT clinical trial for OCU400 for retinitis pigmentosa. Shortly thereafter, the European Medicines Agency (EMA) reviewed the study design, endpoints, and planned statistical analysis, and deemed the US-based trial acceptable for a Marketing Authorisation Application (MAA). In December 2023, the FDA granted OCU400 Regenerative Medicine Advanced Therapy (RMAT) designation.

The Phase III study will include 150 participants75 with the RHO gene mutation and 75 that are gene-agnostic. In each arm, participants will be randomised 2:1 to the treatment group (2.5 x 1010 vg/eye of OCU400) and the untreated control group.

Ocugen plans to expand the OCU400 clinical trial in the second half of 2024 to include patients with LCA, contingent on favourable results from the Phase I/II study and alignment with regulatory agencies.

EPR Podcast 24 Developing modifier gene therapy Ocugen

Unlike conventional methodsmodifier gene therapyemphasises the importance of the broader biological system, potentially leading to more effective treatments.

The gene-agnostic mechanism of action for OCU400 provides hope for a larger retinitis pigmentosa patient population and demonstrates the potential to expand the range of indications for which modifier gene therapy could apply.

Unlike conventional methods that typically focus on replacing a mutated gene with a functional copy, modifier gene therapy modifies gene expression using master gene regulators. These master regulators work in a gene-agnostic way and open the possibility for the treatment of diseases caused by different gene mutations.

Modifier gene therapy triggers epigenetic mechanisms to restore homeostasis in the cellular environment and thereby structural and functional improvement in affected cells.

This pragmatic approach emphasises the importance of the broader biological system, potentially leading to more effective treatments.

OCU410 is a modifier gene therapy for the treatment of geographic atrophy, an advanced stage of dry age-related macular degeneration (dAMD). It utilises an adeno-associated virus (AAV) delivery platform for the retinal delivery of the RAR-related orphan receptor A (RORA).

The RORA protein plays a crucial role in stress and metabolism, reducing lipofuscin deposits and oxidative stress, and demonstrates anti-inflammatory properties as well as inhibiting the complement system in in vitro and in vivo studies.

Ocugen is currently enrolling patients in the Phase I/II ArMaDa clinical trial to assess the safety and efficacy of OCU410 for GA secondary to dAMD.

The ArMaDa clinical trial will assess the safety and efficacy of unilateral subretinal administration of OCU410 in subjects with GA and will be conducted in two phases.

OCU410STis a modifier gene therapy utilising an AAV delivery platform (AAV5) for the retinal delivery of the RORA gene for treating the genetic eye disorder Stargardt disease.

The GARDian clinical trial will assess the safety and efficacy of unilateral subretinal administration of OCU410ST in participants and will be conducted in two phases.

In May 2024, Ocugen announced the second cohort (medium dose) completed dosing in the dose-escalation phase. To date, six patients with Stargardt disease have been dosed in the Phase I/II clinical trial. An additional three patients will be dosed with the high dose (cohort 3) in the dose-escalation phase.

Manufacturing of AAV vectors for gene therapy presents several significant challenges, such as:

By leveraging new technologies, [gene therapy] manufacturers can overcome existing challenges, streamline production workflows, and accelerate the development and commercialisation

There has been advancement in many areas related to gene therapy manufacturing, which has played a crucial role in accelerating gene therapy manufacturing by improving efficiency across the production process. By leveraging new technologies, manufacturers can overcome existing challenges, streamline production workflows, and accelerate the development and commercialisation.

Some of these improvements include:

Biopharmaceuticals, Clinical Development, Clinical Trials, Data Analysis, DNA, Drug Development, Drug Safety, Gene therapy, Industry Insight, Research & Development (R&D), Technology, Therapeutics

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Developing and manufacturing modifier gene therapy - European Pharmaceutical Review