Archive for May, 2024

Paralog meta-analysis

To reanalyze the data from the 5 paralog screens, raw read counts were downloaded, and the same pipeline was applied to all of them. A pseudocount of 5 reads was added to each construct in each replicate, and total read counts were normalized to 500 reads per construct. Log2 fold change (LFC) for each guide at late time point was calculated relative to the plasmid sequence counts.

The data from each study (except Thompson) were divided into three groups; the constructs that target single genes paired with non-essential/non-targeting gRNAs (N) in the first position (gene_N), in the second position (N_gene) and constructs that target gene pairs (A_B). LFC values of each group were scaled individually so that the mode of each group was set to zero. Next, all three groups were merged in one table. Before dividing Itos dataset into three groups, LFC values were scaled such that the mode of negative controls (non-essential_AAVS1) would be zero and also TRIM family was removed from this dataset to avoid false paralog pair discovery13. Since in Thompsons study there was just one position for singleton constructs, LFC values were scaled so that the mode of negative controls (non-essential_Fluc) was set to zero. In the next step, LFC of each construct was calculated by the mean of LFC across different replicates.

To calculate genetic interaction, single gene mutant fitness (SMF) was calculated as the mean construct log fold change of gene-control constructs for each gene. The control was either non-essential genes or non-targeting gRNAs. For each gene pair, the expected double mutant fitness (DMF) of genes 1 and 2 was calculated as the sum of SMF of gene 1 and SMF of gene 2. The difference between expected and observed DMF, the mean LFC of all constructs targeting genes 1 and 2, was called dLFC.

Next step was calculating a modified Cohens D between observed and expected distribution of LFC of gRNAs targeting genes. Expected distribution of gRNAs targeting a gene pair, was calculated using expected mean and expected standard deviation (std).

$${expected},{mean}=mu 1+mu 2$$

(1)

$${expected},{stand},{deviat}=sqrt{{({std}1)}^{2}+{({std}2)}^{2}}$$

(2)

$${S}_{{pooled}}=frac{sqrt{{({expected} , {std})}^{2}+{({observed} , {std})}^{2}}}{2}$$

(3)

$${Cohe}{n}^{{prime} }{sD}=frac{{expected},{mean}-{observed},{mean}}{{S}_{{pooled}}}$$

(4)

Where 1 = mean LFC of gene1 constructs, 2 = mean LFC of gene2 constructs, std1 = standard deviation of LFC of gene1 constructs, and std2 = standard deviation of LFC of gene2 constructs.

In each cell line, the paralog pairs with dLFC<1 and Cohens D>0.8 were selected as hits. Cohens D>0.8 indicates large effect size between two groups, meaning that our expected and observed distribution of gRNAs are meaningfully separated. In total 388 paralog pairs were identified as hits across all the studies.

To identify the most consistent method in terms of hit identification, the Jaccard similarity coefficient of every pair of cell lines in each study was calculated by taking the ratio of intersection of hits over union of hits. For the studies that screened more than two cell lines, the final platform weight was the median of the calculated Jaccard coefficients of all pairs of cell lines.

$$Jleft(A,, Bright)=frac{left|Acap Bright|}{left|Acup Bright|}=frac{left|Acap Bright|}{left|A{{{{{rm{|}}}}}}+left|Bright|-{{{{{rm{|}}}}}}Acap Bright|}$$

(5)

To score paralog pairs, each hit was scored based on the cell lines in which it was identified as a hit; cell lines were weighted based on the platform weight described above. We defined the paralog score as the sum of platform weights of cell lines in which the paralog pair was identified as a hit minus the sum of platform weights of cell lines in which the paralog pair was assayed but not identified as a hit (a miss). The distribution of scores is shown in Fig.1. Gene pairs with paralog score > 0.25 and were identified as a hit in two or more studies were listed as candidate gold standard paralog synthetic lethals.

To construct an all-in-one vector for expression of both Cas12a and a guide array, we first swapped in puromycin resistance in place of blasticidin resistance from pRDA_174 (Addgene #136476). We then tested four locations for the insertion of a U6-guide expression cassette; notably this cassette needs to be oriented in the opposite direction of the primary lentiviral transcript to prevent Cas12a-mediated processing during viral packaging in 293T cells. The construct with the best-performing location, between the cPPT and the EF-1 promoter, was designed pRDA_550 (Addgene #203398). Synthesis of DNA and custom cloning was performed by Genscript.

An oligonucleotide pool consisting of 7 Essential and 7 Non-Essential gene crRNAs with their nearby DR, BsmBI recognition as well as overhang sequence was synthesized by Integrated DNA Technologies. The pool was amplified by asymmetric PCR followed by being assembled into PRDA_550 vector to acquire the designed library through NEBridge Golden Gate Assembly Kit (BsmBI-v2) (New England Biolabs). The assembled product was transformed into NEB Stable Competent E. coli (High Efficiency) cells and the plasmid DNA was purified using the PureLink Plasmid Purification Kit (Invitrogen). Three oligonucleotide pools were cloned separately and pooled together to acquire the final 7mer library. The library was sequenced to confirm uniform and complete library representation.

Human paralogs and percent identity data were imported from BioMart, which reports both AB and BA percent identity (these can differ if the two genes encode proteins of different lengths) Mean percent identity ((AB+BA)/2)and delta percent identity (|ABBA|) between paralogs were then calculated, and for the prototype library, paralogs with mean percent identity between 30% and 99% and delta percent identity <10% were selected (Supplementary Fig.5). Next, CCLE expression data was downloaded, and the mean and standard deviation of expression across all CCLE samples was calculated for each gene. Paralogs where both genes had mean expression>2 and stdev<1.5 were selected (i.e. constitutively expressed genes).

Finally, to identify and include paralog families of size > 2, we applied a difference from top paralog filter. For each gene A in the pool, we identified its top paralog B by max sequence identity. Then for each other candidate paralog C, we calculated the drop in sequence identity, ABAC (see distribution of drop % in Supplementary Fig.5). For the prototype library, we defined A,B,C as being in the same family if ABAC<10%.

For the final Inzolia library, we relaxed several of these filters. The delta percent identity filter and the expression variance filter were removed entirely, and the difference from top paralog filter was expanded to 20%. The mean expression filter was retained. These three filtering steps resulted in a total of 4435 paralog pairs included in the Inzolia pool library.

Oligonucleotide pools consisting of designed four-plex guide arrays were synthesized by Twist Bioscience. The prototype pool consists of 43,972 arrays targeting 19,687 single genes, 2082 paralog pairs, 167 paralog triples, and 48 paralog quads.

5-AATGATACGGCGACCACCGAcgtctcgAGATnnnnnnnnnnnnnnnnnnnnTAATTTCTACTATTGTAGATnnnnnnnnnnnnnnnnnnnnAAATTTCTACTCTAGTAGATnnnnnnnnnnnnnnnnnnnnTAATTTCTACTGTCGTAGATnnnnnnnnnnnnnnnnnnnnTTTTTTGAATggagacgATCTCGTATGCCGTCTTCTGCTTG-3.

Italic: primer sequence Bold: BsmBI restriction sequence. Overhang in CAPS. nnnnn: guide sequence Underlined: DR sequence.

The pool of guide arrays was PCR amplified using KAPA HiFi 2X HotStart ReadyMix (Roche) using 20ng of starting template per 25L reaction (primers are listed in Supplementary Data10) and the following conditions: denaturation at 95C for 3min, followed by 12 cycles of 20s at 98C, 30s at 60C, and 30s at 72C, followed by a final extension of 1min at 72C. The resulting amplicon was purified by the Monarch PCR & DNA Cleanup Kit (New England Biolabs) and cloned into the pRDA-550 vector by NEBridge Golden Gate Assembly Kit (BsmBI-v2) The product from assembly reaction was purified and electroporated into Endura Electrocompetent cells (Lucigen). Transformed bacteria were diluted 1:100 in 2xYT medium containing 100g/mL carbenicillin (Sigma) and grown at 30 C for 16h. The plasmid DNA was extracted by PureLink Plasmid Purification Kit (Invitrogen). The library was sequenced to confirm uniform and complete library representation. The library was prepared in MD Anderson Cancer Center.

The final Inzolia pool consists of arrays targeting 19,687 single genes, 4435 paralog pairs, 376 paralog triples, and 100 paralog quads, plus 20 arrays targeting EGFP, 500 targeting intergenic loci, and 50 encoding non-targeting guides. Each array in the oligonucleotide pools is constructed as follows:

5-AGGCACTTGCTCGTACGACGcgtctcgAGATnnnnnnnnnnnnnnnnnnnnTAATTTCTACTATTGTAGATnnnnnnnnnnnnnnnnnnnnAAATTTCTACTCTAGTAGATnnnnnnnnnnnnnnnnnnnnTAATTTCTACTGTCGTAGATnnnnnnnnnnnnnnnnnnnnTTTTTTGAATggagacgTTAAGGTGCCGGGCCCACAT-3.

Italic: primer sequence Bold: BsmBI restriction sequence. Overhang in CAPS. nnnnn: guide sequence Underlined: DR sequence.

The pool of guide arrays was PCR amplified using NEBNext High-Fidelity 2X PCR Master Mix (NEB) using 196ng of starting template per 50L reaction (primers are listed in Supplementary Data10) and the following conditions: denaturation at 98C for 1min, followed by 7 cycles of 30s at 98C, 30s at 53C, and 30s at 72C, followed by a final extension of 5min at 72C. The resulting amplicon was purified by the Qiaquick PCR Purification Kit (Qiagen) and cloned into the pRDA-550 and pRDA-052 via Golden Gate cloning with Esp3I (Fisher Scientific) and T7 ligase (Epizyme). The assembly product was purified by isopropanol precipitation, electroporated into Stbl4 electrocompetent cells (Life Technologies) and grown at 37 C for 16h on agar with 100 ug/mL carbenicillin. Colonies were scraped and plasmid DNA (pDNA) was extracted via HiSpeed Plasmid Maxi (Qiagen). The library was sequenced to confirm uniform and complete library representation. The library was prepared in Broad institute.

K-562 and A549 cells were a gift from Tim Heffernan. A375 and MELJUSO were obtained from the Cancer Cell Line Encyclopedia. Cell line identities were confirmed by STR fingerprinting by M.D. Anderson Cancer Centers Cytogenetic and Cell Authentication Core. All cell lines were routinely tested for mycoplasma contamination using cells cultured in non-antibiotic medium (PlasmoTest Mycoplasma Detection Assay, InvivoGen).

All cell lines were grown at 37C in humidified incubators at 5.0% CO2 and passaged to maintain exponential growth. For each cell line, the following medium and concentration of polybrene (EMD Millipore) and puromycin (Gibco) were used:

K-562: RPMI+10% FBS, 8g/mL, 2g/mL

A549: DMEM+10%FBS, 8g/mL, 2g/mL

A375: RPMI+10% FBS, 1g/mL, 1g/mL

MELJUSO: RPMI+10% FBS, 4g/mL, 1g/mL.

Lentivirus was produced by the University of Michigan Vector Core (prototype) or the Broad GPP (Inzolia). Virus stocks were not titered in advance. Transduction of the cells was performed at 1X concentration of virus with corresponding polybrene. Non-transduced cells were eliminated via selection puromycin dihydrochloride. The selection was maintained until all non-transduced control cells reached 0% viability. Once selection with puromycin was complete, surviving cells were pooled and 500x coverage cells were harvested for a T0 sample. After T0, cells were harvested at 500X coverage on corresponding days. The prototype In4mer screens were performed in MD Anderson Cancer Center. The Inzolia screens were performed in Broad Institute.

Genomic DNA (gDNA) was extracted using the Mag-Bind Blood & Tissue DNA HDQ 96 Kit (Omega Bio-tek) and quantified by the Qubit dsDNA Quantification Assay Kits (ThermoFisher). Illumina-compatible guide array amplicons were generated by amplification of the gDNA in a one-step PCR. Indexed PCR primers were synthesized by Integrated DNA Technologies using the standard 8nt indexes from Illumina (D501-D508 and D701-D712) (Supplementary Data10).

At least ~200X coverage gDNA per replicate across multiple reactions were amplified. Each gDNA sample was first divided into multiple 50L reactions with most 2.5ug gDNA per reaction. Each reaction contained 1ul each primer (10M), 1L 50X dNTPs, 5% DMSO, 5L 10X Titanium Taq Buffer, and 1L 10X Titanium Taq DNA Polymerase (Takara). The PCR conditions were: denaturation at 95C for 60s, followed by 25 cycles of 30s at 95C and 1min at 68C, followed by a final extension at 68C for 3min. After the PCR, all reactions from the same sample were pooled and then purified by E-Gel SizeSelect II Agarose Gels, 2% (ThermoFisher). Purified amplicons were quantified by Qubit dsDNA Quantification Assay Kits (ThermoFisher) and validated by D1000 ScreenTape Assay for TapeStation Systems (Agilent) (360bp for in4mer, 501bp for 7Mer). Purified amplicons were then pooled (with 30% customized random library to increase the diversity) and sequencing was performed by NextSeq 500 sequencing platform (Illumina) with custom primers (Integrated DNA Technologies) (Supplementary Data10). The In4mer library was sequenced by read format of 151-8-8, single-end and the 7Mer library was sequenced by read format of 151-8-8-151, paired-end.

Genomic DNA (gDNA) was extracted using the Mag-Bind Blood & Tissue DNA HDQ 96 Kit (Omega Bio-tek) and quantified by the Qubit dsDNA Quantification Assay Kits (ThermoFisher). Illumina-compatible guide array amplicons were generated by amplification of the gDNA in a one-step PCR. Indexed PCR primers were synthesized by Integrated DNA Technologies using the standard 8nt indexes from Illumina (D501-D508 and D701-D712). The sequences for the primer sets were listed in Supplementary Data10.

At least ~200X coverage gDNA per replicate across multiple reactions were amplified. Each gDNA sample was first divided into multiple 100L reactions with most 10g gDNA per reaction. Each reaction contained 0.5L forward primer (100M), 10uL reverse primer (5 uM) 8L dNTPs, 5L DMSO, 10L 10X Titanium Taq Buffer, and 1.5L Titanium Taq DNA Polymerase (Takara). The PCR conditions were: An initial denaturation at 95C for 60s, followed by 28 cycles of 30s at 94C, 30s at 52C, and 30s at 72 C followed by a final extension at 72C for 10min. After the PCR, all reactions from the same sample were pooled and purified with Agencourt AMPure XP SPRI beads according to the manufacturer protocol (Beckman Coulter). Purified amplicons were quantified by Qubit dsDNA Quantification Assay Kits (ThermoFisher) and sequenced on a HiSeq2500 with a Rapid Run (200 cycle) kit (Illumina).

Reads for each reagent were counted using only exact matches to the entire 281 nucleotide 7mer sequence, excluding the leading DR (7 23mer spacer sequences + 6 20mer DR sequences). Fold changes were calculated relative to the mean of the T0 samples, and averaged across replicates. For each sample (T7/14/21), fold changes were normalized by subtracting the mean fold change of arrays with 7 nonessentials; i.e. setting no-essentials guides to zero.

We expected that the selected essential genes would not show any pairwise or higher order interactions, and thus should be governed by the multiplicative model of genetic interaction. To evaluate this model, we fit a regression model:

where A is a binary matrix of 7mer guide arrays (rows, k=384) by positions (columns, n=7), with Ai,j=1 if guide array i targets an essential gene at position j and 0 if not. (y) is the vector of normalized observed fold changes, and the n-length vector (beta) coefficients represent the single gene knockout phenotype learned from the model. We filtered this construct for reagents that encoded two or fewer essential genes (k=87 rows). After linear fit, we compared the predicted zero, one, and two gene knockout fitness profiles (by summing the (beta) coefficients for each gene) to the mean observed knockout fitness. R2 values for each pool ranged from 0.78 to 0.91, and the overall quality of the linear fit supports the multiplicative model for non-interacting genes as assayed by combinatorial CRISPR knockouts of up to two genes. An accurate null model for noninteraction is critical for detecting and classifying deviations from this model that reflect positive or negative genetic interactions.

In4mer library sequencing reads were mapped to the library using only perfect matches. BAGEL2 was used to normalize sample level read counts and to calculate fold changes relative to the T0 reference using the BAGEL2.py fc option with default parameters44. Essential and non-essential genes were defined using the Hart reference sets from refs. 39,41. Since the library targets both individual genes and specific gene sets (paralogs), we calculated the average gene/gene set (hereafter gene) log fold change as the mean of the clone-level fold changes across two replicates. All fold changes are calculated in log2 space. Cohens D statistics were calculated in Python as described in Paralog meta-analysis above. Data for recall-precision curves were calculated using BAGEL2. We set an arbitrary threshold of fc<1 for essential genes.

For genetic interaction analysis, the expected fold change was calculated as the sum of the gene-level fold changes for each individual gene in the gene set. Expected fc was subtracted from observed fc to calculate delta log fold change, dLFC, where negative dLFC indicates synthetic/synergistic interactions with more severe negative phenotype, and positive dLFC indicates positive/suppressor/masking interactions with less severe negative or more positive phenotype than expected. We set an arbitrary threshold of dLFC<1 for synthetic lethality, and>+1 for masking/suppressor interactions.

An arrayed knockout apoptosis assay approach was adopted to validate RAS synthetic lethality in K-562. Two guides were selected for each of the three RAS genes, and two clones were designed for each target/gene combination. Guide RNAs were selected through CRISPick and gblocks (same construct as Inzolia library) were synthesized by Integrated DNA Technologies. The arrays were individually cloned into the pRDA_550 backbone and plasmids were validated by Sanger sequencing. The plasmids were then individually transfected to K-562 cells via the Neon Transfection System (Invitrogen). Each group was transfected with 2g of DNA per 2106 cells, using the recommended setting for K-562 electroporation with one pulse at 1000v, 50ms. Non-transfected cells were eliminated through puromycin selection, which was maintained until non-transfected control cells reached 0% viability. Triplicate wells were maintained after selection until the end of the experiment. Cell viability, total cell numbers, live cell size and dead cell size data were collected through reading Trypan Blue (Gibco) stained cells via Countess II FL (Thermo Fisher) at each passage until 9 days after puromycin selection, in line with Inzolia screen end point of 8 days in K-562 cells. Percent dead cells were normalized to negative control and one-way ANOVA was conducted to compare experimental groups against the negative control for statistical significance.

Further information on research design is available in theNature Portfolio Reporting Summary linked to this article.

Visit link:
Efficient gene knockout and genetic interaction screening using the in4mer CRISPR/Cas12a multiplex knockout platform - Nature.com

Rey RA, Grinspon RP, Gottlieb S, Pasqualini T, Knoblovits P, Aszpis S, et al. Male hypogonadism: an extended classification based on a developmental, endocrine physiology-based approach. Am Soc Androl Eur Acad Androl. 2012;1:316.

Google Scholar

Kumar P, Kumar N, Thakur DS, Thakur DS, Patidar A. Male hypogonadism: symptoms and treatment. J Adv Pharm Technol Res. 2010;1:297301.

Article PubMed PubMed Central Google Scholar

Fieldman AH, Longcope C, Derby CA, Johannes CB, Aroujo AB, Coviello AD, et al. Age trends in the level of serum testosterone and other hormones in middle-aged men: longitudinal results from the Massachusetts male aging study. J Clin Endocrinol Metab. 2002;87:58998.

Article Google Scholar

Mulligan T, Frick MF, Zuraw QC, Stemhagen A, McWhirter C. Prevalence of hypogonadism in males aged at least 45 years: the HIM study. Int J Clin Pract. 2006;60:7629.

Article CAS PubMed Google Scholar

Ventimiglia E, Ippolito S, Capogrosso P, Pederzoli F, Cazzaniga W, Boeri L, et al. Primary, secondary and compensated hypogonadism: a novel risk stratification for infertile men. Andrology. 2017;5:50510.

Article CAS PubMed Google Scholar

George M, Yulia T, Svetlana K. Influence of testosterone gel treatment on spermatogenesis in men with hypogonadism. Gynecol Endocrinol. 2014;30:2224.

Article CAS PubMed Google Scholar

Vigen R, ODonnell CI, Barn AE, Grunwald GK, Maddox TM, Bradley SM. Association of testosterone therapy with mortality, myocardial infarction, and stroke in men with low testosterone levels. JAMA. 2013;310:182936.

Article CAS PubMed Google Scholar

Chen L, Wolff DW, Xie Y, Lin M, Tu Y. Cyproterone acetate enhances TRAIL-induced androgen-independent prostate cancer cell apoptosis via up-regulation of death receptor 5. BMC Cancer. 2017;17:11.

Article CAS Google Scholar

Jung M, Oh K, Choi EJ, Oh D, Kim YJ, Bae D. In vitro and in vivo androgen regulation of Dendropanax morbiferus leaf extract on late-onset hypogonadism. Cell Mol Biol (Noisy-le-grand). 2018;64:2027.

Article PubMed Google Scholar

Tripathy A, Ara F, Ghosh P, Ghosh D. Effect of lycopene on testicular androgenic and anti-oxidative enzymes in cyproterone acetate-induced male infertile Wistar strain albino rats: an in vitro study. Andrologia. 2020; https://doi.org/10.1111/and.13494

Nde Z, Massoma LD, Wankeu-Nya M, Ngaha MI, Koloko LB, Bend EF. Potential activity of Aframomum daniellii (Zingiberaceae) dry seeds: a case study of its action mechanism on the Wistar rat strain with testicular deficiency. Biomed Pharmacother. 2020;131:110759.

Article Google Scholar

Rockville MD. US food and drug administration (USFDA). Guidance for industry: estimating the maximum safe starting dose in adult healthy volunteer. 2005

Yakubu, MT, Akanji, MA. Effect of Aqueous Extract of Massularia acuminata Stem on Sexual Behavior of Male Wistar Rats. Evidence-Based Complement Altern Med. 2011; https://doi.org/10.1155/2011/738103

Souza AD, Bessa DH, Fernandes CC, Pereira PS, Martin CH, Miranda ML Phytochemical screening of extracts from Spiranthera odoratissima A. St.-Hil. (Rutaceae) leaves and their in vitro antioxidant and anti-Listeria monocytogenes activities. Acta Sci Biol Sci. 2020; https://doi.org/10.4025/actascibiolsci.v42i1.51881

OECD: Organisation for Economic, Cooperative and Development. Guideline for testing of chemicals, test n423, acute oral toxicity acute toxic class method. 2001; 114

Cariton AE. Experimental surgery of the genital system. In: Gay WI and Heavener JE (eds). Methods of animal experimentation: research surgery and care of the research animal; part B chirurgical approaches to organ systems. Academic Press, Orlando, 1986, pp 191.

Watcho P, Wankeu-Nya M, Nguelefack TB, Tapondjou L, Teponno R, Kamanyi A. Pro-sexual effects of Dracaena arborea (WILD) LINK (Dracaenaceae) in sexually experienced male rats. Pharmacologyonline. 2007;1:40019.

Google Scholar

Ageel AM, Islam MW, Ginawi OT, Al-Yahya, MA. Evaluation of the aphrodisiac activity of Litsea chinensis (Lauraceae) and Orchis malculata (Orchidaceae) extracts in rats. Phyther Res. 1994; https://doi.org/10.1002/ptr.2650080211

Carro-Jurez M, Cervantes E, Cervantes-Mndez M, Rodrguez-Manzo G. Aphrodisiac properties of Montanoa tomentosa aqueous crude extract in male rats. Pharmacol Biochem Behav. 2004;78:12934.

Article PubMed Google Scholar

Chauhan N, Dixit V. Spermatogenic activity of rhizomes of Curculigo orchioides Gaertn in male rats. Int J Appl Res Nat Prod. 2008;1:2631.

Google Scholar

Robb GW, Amann RP, Killian GJ. Daily sperm production and epididymal sperm reserves of pubertal and adult rats. J Reprod Fertil. 1978;54:1037.

Article CAS PubMed Google Scholar

Ernst EA. Method for evaluation of epididymal sperm count and motility in the rat. Scand J Lab Anim Sci. 1989;16:6771.

Google Scholar

Chen C, Gao G, Ho D, Lin C, Chou Y, Chen S. Cyproterone acetate acts as a disruptor of the aryl hydrocarbon receptor. Sci Rep. 2021; https://doi.org/10.1038/s41598-021-84769-7

Zambl A, Sahpaz S, Brunet C, Bailleul F. Effects of Microdesmis keayana roots on sexual behavior of male rats. Phytomedicine. 2008;15:6259.

Article PubMed Google Scholar

Yakubu MT, Akanji MA, Oladiji AT, Adesokan AA. Androgenic potentials of aqueous extract of Massularia acuminata (G. Don) Bullock ex Hoyl. stem in male Wistar rats. J Ethnopharmacol. 2008;118:50813.

Article CAS PubMed Google Scholar

Gauthaman K, Ganesan AP. The hormonal effects of Tribulus terrestris and its role in the management of male erectile dysfunction-an evaluation using primates, rabbit and rat. Phytomedicine. 2008;15:4454.

Article CAS PubMed Google Scholar

Moundipa PF, Beboy NS, Zelefack F, Ngouela S, Tsamo E, et al. Effects of Basella alba and Hibiscus macranthus extracts on testosterone production of adult rat and bull Leydig cells. Asian J Androl. 2005;7:4117.

Article PubMed Google Scholar

Kim JY, Wood RI. Anabolic-androgenic steroids and appetitive sexual behavior in male rats. Horm Behav. 2014;66:58590.

Article CAS PubMed PubMed Central Google Scholar

Shimon I, Lubina A, Gorfine M, Ilany J. Feedback inhibition of gonadotropins by testosterone in men with hypogonadotropic hypogonadism: comparison to the intact pituitary-testicular axis in primary hypogonadism. J Androl. 2006;27:35864.

Article CAS PubMed Google Scholar

Kotta S, Ansari SH, Ali J. Exploring scientifically proven herbal aphrodisiacs. Pharmacogn Rev. 2013;7:110.

Article PubMed PubMed Central Google Scholar

Simanainen U, McNamara K, Davey RA, Zajac JD, Handelsman DJ. Severe subfertility in mice with androgen receptor inactivation in sex accessory organs but not in testis. Endocrinology. 2008;149:33308.

Article CAS PubMed Google Scholar

Grande G, Barrachina F, Soler-Ventura A, Jodar M, Mancini F, Marana R. The Role of Testosterone in Spermatogenesis: Lessons From Proteome Profiling of Human Spermatozoa in Testosterone Deficiency. Front Endocrinol (Lausanne). 2022. https://doi.org/10.3389/fendo.2022.852661

Article PubMed Google Scholar

Kumar GG, Kilari EK, Nelli NB. Oral administration of Turnera diffusa willd. ex Schult. extract ameliorates steroidogenesis and spermatogenesis impairment in the testes of rats with type-2 diabetes mellitus. J Ethnopharmacol. 2023; https://doi.org/10.1016/j.jep.2023.116638.5

Wankeu-Nya M, Florea A, Blici S, Watcho P, Matei H, Kamanyi A. Dracaena arborea alleviates ultra-structural spermatogenic alterations in streptozotocin-induced diabetic rats. BMC Complement Altern Med. 2013. https://doi.org/10.1186/1472-6882-13-71

Wankeu-Nya M, Florea A, Blici , Matei H, Watcho P, Kamanyi A. Cytoarchitectural improvement in Leydig cells of diabetic rats after treatment with aqueous and ethanol extracts of Dracaena arborea (Dracaenaceae). J Tradit Complement Med. 2021;11:18.

Article CAS PubMed Google Scholar

Link:
Aphrodisiac and androgenic effects of the aqueous extract of the roots of Vepris afzelii on cyproterone acetate-induced ... - Nature.com

From an early age, Allison Keeler 05 (biotechnology) always knew she wanted to be a scientist. As an adult, that dream has come true as she is an assistant professor in the Department of Pediatrics at the University of Massachusetts (UMass) Chan Medical School and the lead researcher in the Keeler Lab within the Horae Gene Therapy Center.

Keeler applied early to RIT, out of her high school in eastern Pennsylvania. After visiting the campus and learning the university had one of the few biotechnology programs available, she knew getting hands-on experience in research at RIT was the best path for her future.

Keeler earned her bachelors degree in three years and was able to take advantage of a trip to the Galpagos Islands, where she realized she wanted to become an academic and a professor. After a research position at Duke University, she earned her Ph.D. in biomedical sciences from the UMass Medical School, where she now works. Mentoring and teaching in a lab environment has become her passion.

Basics that I learned at RIT and each of my experiences have shaped where I am now and what Im really passionate about, said Keeler.

Her background has led her to one of the most revolutionary medical fields today: gene therapy. This technology approaches disease differently, by attempting to change genetic makeups to prevent and treat disease instead of traditional treatments like medication and surgery.

In her lab, Keeler is learning about and understanding immune responses to gene therapy and engineering and developing new novel gene therapies for the treatment of different diseases.

The field is progressing rapidly. When she was a graduate student, there were no approved therapies. Now, there are many, with more being approved every year.

Its been really interesting to watch the field evolve, said Keeler. Its an exciting time in this field because several gene therapies have recently been approved.

The scientific area is familiar to the dean of RITs College of Science, Andr Hudson, who is repeatedly sought out as an expert in biochemistry and microbiology. His research interests are closely related to Keelers, as both are excited about the future of science as it relates to the human body and disease.

The work by Dr. Keeler and colleagues in this space is at the forefront of science and medicine, said Hudson. I am heartened that one of our College of Science alumni is helping to lead the charge.

Keeler never envisioned she would be running her own gene therapy lab when she stepped on RITs campus as a biotechnology major. But as science grows and evolves, more and more possibilities for careers in science exist. She encourages students to keep their minds open and to explore all opportunities.

I didnt even know about gene therapy when I was at RIT, said Keeler. But science continues to expand. Keep being curious, keep asking questions.

See the original post:
Scientist's RIT experience leads to career in revolutionary gene therapy research | RIT - Rochester Institute of Technology

Want to stay on top of the science and politics driving biotech today?Sign upto get our biotech newsletter in your inbox.

Good morning! A crowdsourced Readout today, to be sure, with contributions from STATs Jason Mast, Elaine Chen, and Jonathan Wosen. We get into earnings from Novo Nordisk, Amgen, and Illumina, and see that ICER is not so enthused about a gene therapy from Sarepta.

Also: Wed like to know what you think of this newsletter. if you havent, could you fill outthis survey?

Novo Nordisk CEO Lars Fruergaard Jrgensen is standing by the cost of Ozempic and Wegovy, despite an ongoing Senate investigation into the companys pricing. In an earnings call yesterday, he said the drugs offer an attractive value proposition as theyre priced similarly to earlier, less effective iterations of this class of drugs.

Although he conceded that the number of people taking the drugs is to some degree putting strains on health care systems, STATs Elaine Chen writes, the full value of these diabetes and obesity drugs has yet to be realized. Ozempics list price is $969 monthly; Wegovys is $1,349. Executives on the call said that the net prices for these drugs have already come down, and will continue to do so as more people take it and as competition increases.

Read more.

The removal of small amounts of brain tissue from desperately ill patients, done as part of a Mount Sinai research project, triggered alarm bells at the FDA and has raised broader questions about the scientific and ethical justification for live-brain research. Journalist and STAT contributor Katherine Eban joins The Readout LOUD this week to discuss the findings of atwo-year investigation.

Also on this weeks episode, STATs Adam Feuerstein and Allison DeAngelis discuss Novartis effort to acquire MorphoSys, and the latest news on Eli Lilly and Novo Nordisks blockbuster obesity drugs with Elaine Chen.

Listen here.

From STATs Jason Mast:The drug-pricing watchdogs over at the Institute for Clinical and Economic Review, or ICER, often take a dim view of modern drug pricing exceptwhen it comes to genetherapy. Those treatments, the nonprofit has said, often provide the years-long benefits for severe disease that justify a multimillion-dollar price tag.

Which makes theJAMApiecepublishedthis week by ICER CMO David Rind all the more notable. Rind considered Elevidys, Sareptas $3.2 million gene therapy for Duchenne muscular dystrophy. The treatment was given accelerated approval last year for 4- and 5-year-olds and, after a Phase 3 trial, the FDA is now considering whether to approve it for all ages.

That Phase 3 trial missed its primary endpoint, though, as did a previous smaller study. Sarepta has pointed to the results on secondary measures but given those data, Rind cast doubt on whether it should be approved and certainly whether Sarepta should charge what other gene therapy companies do. This is an enormous price tag for a therapy that has failed to meet its primary end point in the 2 randomized trials in which it has been studied and that is clearly not curative, he wrote.

From STATs Jonathan Wosen:DNA sequencing juggernaut Illumina reported yesterday $1.06 billion in revenue for its core business during the first quarter of this year, down 2% from the same time last year, with the companys execs reiterating that they expect 2024 revenue to essentially match the $4.5 billion from the previous fiscal year.

CEO Jacob Thaysen cautiously described the first-quarter numbers as a decent start to the year that exceeded expectations on a call with market analysts and investors. The updated numbers come during a turbulent time for the broader sequencing space. Pacific Biosciences, a Bay Area company that Illumina once unsuccessfully tried to acquire, recently laid off nearly 200 employees after reporting disappointing first-quarter sales. And while Illumina continues to control about 80% of the market, its shares are down 37% from a year ago and the firm faces growing competition from players such asUltima, Singular Genomics, andElement Biosciences.

The San Diego genomics giant is still on track to finalize the terms of its divestiture of Grail a cancer detection startup Illumina had acquired for $8 billion, drawing the ire of regulators in the U.S. and Europe by the end of the second quarter of this year.

From STATs Elaine Chen:Amgen will scrap an early-stage obesity pill, and will instead focus on its more advanced injectable candidate called MariTide thats seen as a potential competitor to Wegovy and Zepbound.

MariTide is in a Phase 2 trial for obesity, and CEO Bob Bradway said on the earnings call yesterday that we are very encouraged with the results from an interim analysis of that trial. The company is planning a broad Phase 3 program that will study MariTide in obesity, diabetes, and obesity-related conditions, and its already gearing up to make large amounts of the drug, initiating activities to further expand manufacturing capacity with both clinical and commercial supply in mind, Bradway said.

MariTide has an interesting mechanism its a monoclonal antibody linked to two peptides that activates receptors of the GLP-1 hormone while blocking receptors of the GIP hormone. Even though that appears to contradictory to the mechanism of Lillys potent obesity drug Zepbound, which activates both GLP-1 and GIP receptors, MariTide has shown potential to induce potent and longer-lasting weight loss.

Read more.

See the rest here:
Readout Newsletter: Amgen, Illumina, Novo Nordisk, and more - STAT

Electrical engineering PhD student Yizhou Yao had to develop a whole new set of skills to research the effects of electric pulses on the receptivity of liver cells to new types of gene therapy. Photo by Joel Hallberg

In an effort to improve delivery of costly medical treatments, a team of researchers in electrical engineering at the University of WisconsinMadison has developed a stimulating method that could make the human body more receptive to certain gene therapies.

The researchers exposed liver cells to short electric pulses and those gentle zaps caused the liver cells to take in more than 40 times the amount of gene therapy material compared to cells that were not exposed to pulsed electric fields. The method could help reduce the dosage needed for these treatments, making them much safer and more affordable. The research appears April 30 in the journal PLOS ONE.

Gene therapy is a promising medical technology: By replacing, altering or introducing new genetic material into a patients cells, doctors may be able to cure or compensate for genetic diseases, including cystic fibrosis, sickle-cell disease, hemophilia and diabetes.

One of the bottlenecks in gene therapy, however, is getting the right dose of genetic material into the target cells. The UWMadison research suggests that applying a moderate electric field, which left no lasting damage to the cells that received it, could help in creating more effective therapies.

The project began almost a decade ago with Hans Sollinger, a world-renowned transplant surgeon at UWMadison. He had developed a gene therapy treatment for Type 1 diabetes, an autoimmune disease that attacks the pancreas, the organ that produces insulin.

From left, Professor John Booske, PhD student Yizhou Yao and Professor Susan Hagness, all in the UWMadison Department of Electrical and Computer Engineering. Photo by Joel Hallberg

Sollingers treatment strategy delivered the genetic code for insulin production into liver cells using an adreno-associated virus that assists in transporting the therapeutic genes across the cells membrane. This DNA can then take up residence in liver cells, producing insulin without being attacked by the immune system in the pancreas.

While Sollinger had a proof of concept that the therapy worked, he believed the future of the treatment hinged on delivery. He turned to Susan Hagness and John Booske, both UWMadison professors of electrical and computer engineering who have experience treating human cells with electrical pulses.

What we started talking about was local, targeted delivery and whether there was a way of getting the treatment DNA directly into the liver without passing it through the entire body and triggering the immune system, says Hagness. And whether we could use electric pulses in order to make this delivery process more efficient and dramatically reduce the dose needed.

Researchers have previously found that exposing cells to electric fields can often increase the ability of molecules to move through the cell membrane into the interior of a cell. So, in this latest study, PhD student Yizhou Yao sought to determine whether the technique would increase the penetration of virus particles into liver cells.

Using human hepatoma cells, a model system for studying the liver, Yao exposed batches of the cells to various concentrations of the gene therapy virus particles containing a fluorescent green protein. She used a pair of electrodes to deliver an 80-millisecond electric pulse to some samples, then incubated all the cells for 12 hours.

When she examined the results 48 hours later under a fluorescence microscope, Yao found that only a small percentage of the cells that had not received the electrical pulses glowed green. In stark contrast, those cells that had received a zap accumulated about 40 times the amount of the fluorescent green proteins delivered by the virus.

While results provided compelling evidence that the pulses helped facilitate the viruss penetration of the cell walls, Booske says the team has yet to discover exactly how the process works at the molecular level.

Theres enough known about electric pulsing that I think we could confidently state that it is opening nanopores through the cell membrane, he says. But then Yao got this remarkable result, and it dawned on us that virus particles are in general bigger and more complex than bare molecular particles and they already have their own way of getting inside cells. So, we dont really know if its the pores opening that has anything to do with it directly or indirectly.

Sollinger passed away in May 2023, but the team says his legacy will live on through the ongoing research on this project and the work of other groups. The electrical engineering researchers are pursuing next steps with external funding and are optimistic that ultimately the technique will translate into clinical trials.

Yao, who will graduate in 2024, says she knew the study would be transdisciplinary, but didnt realize just how far it would go.

I am an electrical engineer by training, and I dont have a biology background, she says. Before this, the last time I used a microscope was in high school. It was quite a steep learning curve, learning to culture cells and carry out biology protocols. But I really enjoyed this project and liked its ultimate goal, which is to make the world a better place.

Other authors include Robert W. Holdcraft of the Cincinnati Childrens Hospital Medical Center.

Original post:
An electrifying discovery may help doctors deliver more effective gene therapies - University of Wisconsin-Madison

SMN-K186R, a modified version of the SMN protein that is deficient in people with spinal muscular atrophy (SMA), was more effective than normal SMN when delivered by gene therapy, a mouse study demonstrated.

Treatment with lower doses of the modified SMN gene therapy, to reduce liver-related toxicity seen with current gene therapies, was still more effective than normal SMN.

[SMN-K186R] has value as a new treatment for SMA that improves treatment effectiveness and reduces adverse events simultaneously, the scientists wrote.

The study, Improved therapeutic approach for spinal muscular atrophy via ubiquitination-resistant survival motor neuron variant, was published in the Journal of Cachexia, Sarcopenia and Muscle.

SMA is caused by defects in the SMN1 gene, which leads to a deficiency in the SMN protein. A lack of SMN mainly affects motor neurons, the specialized nerve cells that control movement, resulting in muscle weakness and atrophy (shrinkage) over time.

Zolgensma (onasemnogene abeparvovec-xioi) is a gene therapy widely approved for newborns and toddlers up to age 2 with all types of SMA. Delivered to cells using a modified and harmless adeno-associated virus serotype 9 (AAV9), the therapy is designed to replace the faulty SMN1 gene and increase the levels of SMN protein.

Although clinical trials have shown that Zolgensma can improve motor skills and extend survival, some patients still fail to achieve motor milestones despite treatment before symptom onset. Moreover, in clinical trials some patients showed liver toxicity due to high AAV9 dosage, even after treatment with immunosuppressants.

Therefore, drug development for an improved therapeutic effect for SMA patients is still needed, the scientists wrote.

In cells, proteins are routinely produced and then degraded in an ongoing dynamic process, depending on cellular needs. Proteins targeted for degradation are tagged with a small protein called ubiquitin, a process called ubiquitination.

In previous work, the research team created a mutant version of the SMN protein, called SMN-K186R, that was resistant to ubiquitination but still retained its function in motor neurons. As a result, the levels of SMN-K186R were sustained for longer periods compared with normal SMN (SMN-WT).

Researchers have now developed an AAV9-based gene therapy, similar to Zolgensma, that delivers the genetic instructions for SMN-K186R to cells and tested its efficacy in a mouse model of severe SMA.

Control SMA mice treated with a mock therapy had an average lifespan of 14 days and survived about four weeks following treatment with SMN-WT. In contrast, mice given SMN-K186R lived an average of 164 days, representing an average lifespan extension of more than 140 days, about 10 times longer than without treatment.

Body weights paralleled the survival results. Untreated mice, or those treated with SMN-WT, initially gained weight and then lost weight until death, whereas mice given SMN-K186R gained weight and size comparable to healthy mice.

Tissue analysis found higher levels of SMN protein in the brain, spinal cord, and muscles in SMA mice treated with SMN-K186R compared with SMN-WT, a result related to ubiquitination resistance. Higher SMN levels due to SMN-K186R coincided with more motor neurons in the spinal cord, greater muscle mass, and thicker muscle fibers.

Several motor function tests showed the improved efficacy of SMN-K186R over SMN-WT. In response to gravity, mice treated with SMN-K186R turned their bodies upward successfully, unlike SMN-WT-treated mice, which failed to turn upwards beyond day 23. Hind-limb clasping upon tail suspension occurs in mice with motor deficits: whereas mice treated with SMN-K186R rarely showed this behavior, SMN-WT-treated mice displayed hind-limb clasping for more than 20 out of 30 seconds.

Due to the potential of treatment-related liver failure (hepatotoxicity) associated with gene therapy, the team examined the effects of SMN-K186R on the liver. By day 25, the average liver weight of mice treated with SMN-K186R was within the normal range, whereas SMN-WT-treated mice had significantly lighter livers.

While the number of liver cells was similar between the two groups, only mice treated with SMN-K186R showed liver cell growth comparable to that of healthy mice. Moreover, liver cells from SMN-WT-treated mice showed signs of SMN protein aggregation, which has been reported to cause defects in motor neurons.

When the dose of SMN-K186R was lowered to one-third of the clinical viral dose to reduce liver toxicity, SMN-WT-treated mice displayed hind-limb clasping, whereas mice treated with the lower dose of SMN-K186R showed no signs of motor deficits.

The higher stability and levels of SMN-K186R generated over SMN-WT did not cause neurotoxicity in the brain and spinal cord or showed early signs of neurotoxicity when delivered directly into the bloodstream.

Our research suggests that an improved therapeutic approach for SMA via ubiquitination-resistant SMN, [SMN-K186R], will achieve better therapeutic effects in severe SMA newborn patients, the scientists wrote. The enablement of low AAV dose treatment from the improved treatment effects of [SMN-K186R] provides strong foundations for clinical applications in SMA patients to reduce hepatotoxicity.

Read more here:
Modified SMN protein aids gene therapy efficacy: Mouse study - SMA News Today

LONDON In 2021, a team of Italian researchers reported that an experimental gene therapy they had developed seemed to be correcting the metabolic issues at the core of a rare genetic disease that left children unable to break down sugar molecules.

There remained an open question, however, about whether the therapy could address a particularly debilitating manifestation of the disease, the severe skeletal deformities it caused. Patients with the disease, called Hurler syndrome, suffer from short stature, spinal defects, and extremely stiff joints, complications that greatly limit their quality of life.

But in a new paper, the research team showed that the gene therapy, when delivered to toddlers, successfully staved off those problems for years. The children grew to heights within average norms, and had far more flexible shoulder, hip, and knee joints than untreated children, according to the study, published Wednesday in the journal Science Translational Medicine. Defects in the patients spines also stabilized.

STAT+ Exclusive Story

Already have an account? Log in

Already have an account? Log in

Already have an account? Log in

Get unlimited access to award-winning journalism and exclusive events.

Read more here:
Experimental gene therapy seems to alleviate skeletal defects tied to rare inherited disease, study shows - STAT

Summary: Researchers made a significant breakthrough in ALS treatment using a new gene therapy, marking a notable slow in disease progression for a patient with an aggressive form of ALS. The patient, treated since early 2020, has maintained much of their physical and social abilities, exceeding typical life expectancy and functionality projections for their condition.

This therapy targets the SOD1 gene mutation, reducing levels of a harmful protein and stabilizing the patients condition. The findings offer hope for future advancements in ALS treatment and broader applications of gene therapy.

Key Facts:

Source: Umea University

There has been a breakthrough in the research on the disease amyotrophic lateral sclerosis (ALS).

Scientists at Ume University report that the disease progression in a patient with a particularly aggressive form of ALS disease has slowed down considerably with the use of a new gene therapy.

After four years on the medication, the patient can still climb stairs, rise from his chair, eat and speak well, and live an active and socially fulfilling life.

I consider this a breakthrough for the research we have conducted for more than 30 years, here at Ume University and University Hospital of Northern Sweden. We have never before seen treatment results as effective as these, using any other treatment, says Peter Andersen, a neurologist and professor at the Department of Clinical Sciences at Ume University.

An important discovery is that it is now possible to considerably reduce the levels of the disease-causing SOD1 protein, and simultaneously measure a clear inhibitory effect on furtherdisease progression.

When we diagnosed the patient at the neurology ward inearly spring2020, the patients prognosis was 1.52 years of survival at best. The patient has far, far exceeded expectation.

The patient is from a family in southern Sweden with a particularly aggressive form of ALS disease caused by a mutation in the SOD1 gene. When a relative was diagnosed with ALS, the patient left ablood samplefor research purposes to the ALS research team at Ume University but chose to not learn about the results of the genetic test.

However, the patient was a carrier of the disease gene, and after experiencing muscle weakness four years ago, the patient realized that he too was afflicted. The patient was immediately received by the medical team at University Hospital of Northern Sweden and was diagnosed with early stage ALS disease.

Since the summer of 2020, the patient has been a participant in the Phase III study evaluating a new gene therapy developed for patients with SOD1 mutations causing misfolding and aggregation of SOD1 protein in motorneurons.

Every four weeks, the patient received theexperimental treatmentat a university hospital in Copenhagen in Denmark.

At the time of diagnosis in 2020, the patients levels of the substance neurofilament La biomarker indicating breakdown of nerve cellswas very high. Now, four years later, the levels are reduced by almost 90%.

When the patient was diagnosed at University Hospital of Northern Sweden in April 2020, we measured the level of neurofilament L to be as high as 11,000 nanograms per liter, which is high even for an ALS patient.

In the most recent sample, after 50 injections of the new drug, the level is down to 1,200 to 1,290, which is a substantial decrease of the disease indicator, says Peter Andersen.

The normal level for a person in the patients age group is below 560. In blood, the level of neurofilament has fallen back to normal levels, and was down to 12 during the latest hospital visit. The normal level is less than 13.

The patients level of function, measured using the scale ALSFRSR, is reduced compared to a healthy individual (48 points) but has stayed at almost the same level, around 35 to 37 points, for the last 18 monthsthat means that the patients functional level is reduced by approximately 26% compared to a healthy individual.

A person with this aggressive type of ALS gene mutation that the patient has typically loses 11.5 points every month. That means that without treatment, the expected disease progression would have been very fast and given rise to substantial disability within 612 months, and, most likely, have lead to the patients death in 2021.

That this patient, more or less unimpeded, still can climb stairs four years after disease onset, that is somewhat of a miracle to see, says Karin Forsberg, a neurologist and researcher at the Department of Clinical Sciences who works alongside Peter Andersen and has researched SOD1 and ALS for more than two decades.

To have succeeded with adrug treatmentin this way is a great success and an inspiration. But it does not in any way mean that the job is done. This is just the beginning.

It is also important to remember that the drug in question does not constitute a curative treatment, but it seems able to put the brake on disease progression. It gives us great hope to further develop pharmaceutical treatments for ALS-patients.

There are many types of ALS disease, and only 2% to 6% has an ALS disease caused by a mutation in the SOD1 gene. Many have a familial form of the disease, but mutations in SOD1 have also been found in so-called sporadic cases of ALS.

Whether this drug has a similar effect on other types of ALS disease is currently unknown. There is need for much more research on the subject, says Peter Andersen.

The patient can still do almost all things that he could do when he first joined the study in the summer of 2020his speech is unaffected, and he manages to do everything himself, he mows the lawn, goes shopping, and takes care of his children. Mentally he also feels a lot better, mainly because he now dares to feel hope.

The study that the patient is participating in ends this summer. The medication is not yet available in Sweden, but it has been approved by the United States Food and Drug Administration, FDA, and on the 23 of February 2024 the European Medicines Agency, EMA, recommended the use of the drug on patients with SOD1 gene mutations within the European Union.

However, the New Therapies Council i Sweden has asked the regional health care providers not to prescribe the drug until a health economic evaluation has been provided by the Dental and Pharmaceutical Benefits Agency.

Our next step is to study the results from the patients receiving this drug. It has worked for some, but not all have seen the same positive effect. It could be a question of dosage, or at which disease stage the treatment was initiated.

Maybe additional drugs are required to completely stop the process? Those are questions we now have to try and answer. This is only the beginning, says Karin Forsberg.

She pictures a future where treatment will be given based on what type of ALS disease the patient has, and that it most likely will require a combination of drugs.

She emphasizes that there is much research being conducted both in Sweden and internationally to find new drug targets so that equivalent drugs can be developed for patient groups with other types of ALS, and she is hopeful that it will come true.

We can measure in samples collected from the patient that the disease process is ongoing, but the patients body seems able to compensate. Even now, four years after the patient started taking this new gene therapy drug.

The Swedish Ethical Review Authority approved participation in these studies and now, several years later, we, as well as ALS physicians in other participating countries, see a clear clinical effect on many treated patients, says Peter Andersen.

The next step will be to get approval from the Swedish Ethical Review Authority to study the compensatory mechanisms that treatment with this drug seems to have activated. There might be an opportunity here to get insights into how previously unknown parts of the nervous system work, and to develop even better new drugs.

Author: Peter Andersen Source: Umea University Contact: Peter Andersen Umea University Image: The image is credited to Neuroscience News

Original Research: The findings will appear in eLife.

Go here to read the rest:
Gene Therapy Slows ALS Progression - Neuroscience News

Poseida Therapeutics and Astellas, under its Xyphos Biosciences subsidiary, have initiated a research collaboration and licensing agreement to develop allogeneic cell therapy programs by combining their innovative cell therapy platforms and technologies.

The companies plan to combine Poseida's proprietary allogeneic chimeric antigen receptor T-cell (CAR-T) platform with Xyphos' Accel platform (which combines its convertibleCAR and proprietary MicAbodies to target tumor cells) to create a Poseida-developed CAR-T construct. This CAR-T construct will then be used to develop two convertibleCAR product candidates targeting solid tumors.

Under the terms of the agreement, Poseida will receive $50 million upfront, plus potential development and sales milestones and contingency payments of up to $550 million in total. Additionally, Poseida is eligible for royalties as a percentage of net sales.

Xyphos will reimburse Poseida for costs incurred as part of the research agreement and will be responsible for the development and future commercialization of the products generated from the collaboration.

In August 2023, Astellas invested $25 million for an 8.8% stake in Poseida and paid an additional $25 million for the right to exclusive negotiation and first refusal to license Poseidas P-MUC1C-ALLO1 CAR-T cell therapy product candidate.

Read the original:
Astellas and Poseida to collaborate on oncology cell therapies - LabPulse

Research Triangle Park, N.C., May 02, 2024 (GLOBE NEWSWIRE) -- Asklepios BioPharmaceutical, Inc. (AskBio), a gene therapy company wholly owned and independently operated as a subsidiary of Bayer AG, will deliver nine presentations offering insights into its adeno-associated virus (AAV) research and development, a key area of gene therapy focus for the company, at the American Society of Gene and Cell Therapy (ASGCT) 27th Annual Meeting, which takes place May 711, 2024, in Baltimore, Maryland, USA. Presentations will focus on AAV immune-mediated responses as well as the results from the ongoing Phase 1 clinical trial of AB-1002 gene therapy in patients with advanced heart failure.

Luke Roberts, MBBS, PhD, Medical Director for Clinical Development at AskBio, will deliver an oral presentation sharing new clinical data from the companys ongoing Phase 1 trial of AB-1002 in patients with advanced heart failure. This follows AskBios recent news that AB-1002 was granted FDA Fast Track Designation for the treatment of congestive heart failure (CHF). AB-1002 (also known as NAN-101) is an investigational gene therapy that has not yet received marketing authorization, and its efficacy and safety have not been established or fully evaluated. AskBio previously communicated that the delivery of AB-1002 was well tolerated and resulted in positive preliminary efficacy outcomes in some patients with non-ischemic CHF and may validate that the AAV2i8 vector capsid used is highly cardiotropic when administered as a single intracoronary infusion at relatively low doses.

Preliminary data from the Phase 1 trial of AB-1002 were presented at the 2023 American Heart Association Scientific Sessions in November, and AskBio began enrolling patients in its Phase 2 GenePHIT study of AB-1002 in adults with non-ischemic cardiomyopathy and New York Heart Association (NYHA) Class III heart failure symptoms in January 2024.

AskBios ASGCT presence will also include key presentations showcasing the companys continued commitment to optimizing AAV as a gene therapy, with a focus on preventing or reducing AAV immune response-related adverse events, which remains a vital area of investigation across the gene therapy treatment landscape. Attendees can look forward to an ASGCT spotlight speaker presentation on current immune modulation strategies given by Shari Gordon, PhD, Senior Director of Immunology at AskBio, on Day 3. On Day 4, Shari Gordon will deliver on behalf of Audry Fernandez, PhD, Principal Scientist & Group Lead, Immunology R&D at AskBio, an oral presentation on pre-clinical research into AAV-specific immune responses, and Liujiang Song, PhD, Principal Scientist for R&D Capsid and Biology at AskBio, will offer insights into AAV biology and vector intracellular fate during an oral presentation on AAV episome configuration using third generation long-read sequencing technologies and advanced bioinformatics.

Our presence at ASGCT this year highlights our continued commitment to sharing AAV developments with the gene therapy community. Covering clinical and pre-clinical research, our presentations show our robust progress and ongoing ambition to bring to patients transformative therapies that were once unthinkable, said Gustavo Pesquin, Chief Executive Officer, AskBio. With our clinical and early-stage programs advancing, these are exciting times at AskBio.

With an ambitious portfolio of gene therapies at various stages of research and development, AskBio continues to develop AAV-based therapies to treat some the worlds most debilitating diseases, including CHF, Huntingtons disease, limb-girdle muscular dystrophy, multiple system atrophy, Parkinsons disease, and Pompe disease. By targeting these therapy areas, AskBio aims to deliver breakthrough treatments that could benefit more than an estimated 35 million patients worldwide.17

AskBios presentations at ASGCT include:

About AskBio

Asklepios BioPharmaceutical, Inc. (AskBio), a wholly owned and independently operated subsidiary of Bayer AG, is a fully integrated gene therapy company dedicated to developing life-saving medicines and changing lives. The company maintains a portfolio of clinical programs across a range of neuromuscular, central nervous system, cardiovascular, and metabolic disease indications with a clinical-stage pipeline that includes therapeutics for congestive heart failure, Huntingtons disease, limb-girdle muscular dystrophy, multiple system atrophy, Parkinsons disease, and Pompe disease. AskBios gene therapy platform includes Pro10, an industry-leading proprietary cell line manufacturing process, and an extensive capsid and promoter library. With global headquarters in Research Triangle Park, North Carolina, and European headquarters in Edinburgh, Scotland, the company has generated hundreds of proprietary capsids and promoters, several of which have entered pre-clinical and clinical testing. An early innovator in the gene therapy field, with over 900 employees in five countries, the company holds more than 800 patents and patent applications in areas such as AAV production and chimeric capsids. Learn more at http://www.askbio.com or follow us on LinkedIn.

About Bayer

Bayer is a global enterprise with core competencies in the life science fields of health care and nutrition. In line with its mission, Health for all, Hunger for none, the companys products and services are designed to help people and the planet thrive by supporting efforts to master the major challenges presented by a growing and aging global population. Bayer is committed to driving sustainable development and generating a positive impact with its businesses. At the same time, the Group aims to increase its earning power and create value through innovation and growth. The Bayer brand stands for trust, reliability and quality throughout the world. In fiscal 2023, the Group employed around 100,000 people and had sales of 47.6 billion euros. R&D expenses before special items amounted to 5.8 billion euros. For more information, go towww.bayer.com.

About Viralgen Vector Core

Viralgen is a Contract Development and Manufacturing Organization (CDMO) founded in 2017 and exists as an independently operated subsidiary of AskBio, which is wholly owned and independently operated as a subsidiary of Bayer AG. As a manufacturer of Current Good Manufacturing Practice (cGMP) certified AAV, Viralgen offers the Pro10 based suspension manufacturing platform, a technology licensed from AskBio and developed by Chief Technical Officer Josh Grieger, PhD, and Co-Founder R. Jude Samulski, PhD, at University of North Carolina. The Pro10 platform has been found to increase scalability, performance, and yield of AAV therapies.8 Located in Spain, in the Gipuzkoa Science and Technology Park, Viralgen produces AAV gene therapy treatments for pharmaceutical and biotech companies with the aim of accelerating the delivery of new treatments that may improve patients lives.

The companys clinical facilities have four cGMP manufacturing suites, with 250-liter and 500-liter bioreactors. In 2020, Viralgen expanded within the Scientific Park by constructing a new building with three modules for large-scale commercial manufacturing. Each module of the state-of-the-art space includes three cGMP suites with a manufacturing capacity of >2,000 liters. The first module, which includes a suite dedicated to fully automated fill and finish operations, has received cGMP certification by the Spanish Agency for Medicines and Medical Devices (AEMPS) as part of the EMA network. For more information, visit viralgenvc.com.

Bayer Forward-Looking Statements

This release may contain forward-looking statements based on current assumptions and forecasts made by Bayer management. Various known and unknown risks, uncertainties and other factors could lead to material differences between the actual future results, financial situation, development or performance of the company and the estimates given here. These factors include those discussed in Bayers public reports which are available on the Bayer website atwww.bayer.com. The company assumes no liability whatsoever to update these forward-looking statements or to conform them to future events or developments.

AskBio Forward-Looking Statements

This press release contains forward-looking statements. Any statements contained in this press release that are not statements of historical fact may be deemed to be forward-looking statements. Words such as believes, anticipates, plans, expects, will, intends, potential, possible, and similar expressions are intended to identify forward-looking statements. These forward-looking statements include, without limitation, statements regarding AskBios clinical trials. These forward-looking statements involve risks and uncertainties, many of which are beyond AskBios control. Known risks include, among others: AskBio may not be able to execute on its business plans and goals, including meeting its expected or planned clinical and regulatory milestones and timelines, its reliance on third-parties, clinical development plans, manufacturing processes and plans, and bringing its product candidates to market, due to a variety of reasons, including possible limitations of company financial and other resources, manufacturing limitations that may not be anticipated or resolved in a timely manner, potential disagreements or other issues with our third-party collaborators and partners, and regulatory, court or agency feedback or decisions, such as feedback and decisions from the United States Food and Drug Administration or the United States Patent and Trademark Office. Any of the foregoing risks could materially and adversely affect AskBios business and results of operations. You should not place undue reliance on the forward-looking statements contained in this press release. AskBio does not undertake any obligation to publicly update its forward-looking statements based on events or circumstances after the date hereof.

References

[1] Balestrino R, Schapira AHV. Parkinson disease. Eur J Neurol. 2020;27(1):27-42.

[2] World Health Organization. Parkinson Disease. Available at:https://www.who.int/news-room/fact-sheets/detail/parkinson-disease. Accessed April 2024.

[3] Medina A, et al. Prevalence and Incidence of Huntington's Disease: An Updated Systematic Review and Meta-Analysis. Mov Disord. 2022;37(12):2327-2335.

[4] Malik A, et al. Congestive Heart Failure. In: StatPearls. Treasure Island (FL): StatPearls Publishing; November 7, 2022.

[5] MedlinePlus Genetics. NIH. Limb-girdle muscular dystrophy. Available at:https://medlineplus.gov/genetics/condition/limb-girdle-muscular-dystrophy/#frequency. Accessed April 2024.

[6] Goh Y, et al. Multiple system atrophy [published online ahead of print, 2023 Mar 16]. Pract Neurol. 2023; practneurol-2020-002797.

[7] Stevens D, et al. Pompe Disease: a Clinical, Diagnostic, and Therapeutic Overview. Curr Treat Options Neurol. 2022;24(11):573-588.

[8] Grieger JC, Soltys SM, Samulski RJ. Production of Recombinant Adeno-associated Virus Vectors Using Suspension HEK293 Cells and Continuous Harvest of Vector From the Culture Media for GMP FIX and FLT1 Clinical Vector. Mol Ther. 2016;24(2):287-297.

See the original post:
AskBio Announces Nine Presentations at American Society of Gene and Cell Therapy 27th Annual Meeting 2024 - GlobeNewswire

SOLANA BEACH, Calif., May 02, 2024 (GLOBE NEWSWIRE) -- ClearPoint Neuro, Inc. (Nasdaq: CLPT) (the Company), a global device, cell, and gene therapy-enabling company offering precise navigation to the brain and spine, today announced it will present novel research and exhibit at the 27th American Society of Gene & Cell Therapy Annual Meeting to be held in Baltimore, MD from May 7-11.

The following original research abstracts will be presented at the Poster Session:

Additionally, the Companys technology will be featured in multiple partner posters and presentations. Conference attendees may visit the ClearPoint Neuro booth, #1033, to obtain a comprehensive list. Attendees may also pre-book meetings with our team directly here.

The ASGCT Annual Meeting provides an extraordinary opportunity for us to connect with our existing partners, and attract potential new customers, stated Jeremy Stigall, Chief Business Officer at ClearPoint Neuro. This year, in addition to the posters listed above, there are more than 10 partner-authored abstracts that are enabled by ClearPoint Neuro innovations.

About ClearPoint Neuro

ClearPoint Neuro is a device, cell, and gene therapy-enabling company offering precise navigation to the brain and spine. The Company uniquely provides both established clinical products as well as pre-clinical development services for controlled drug and device delivery. The Companys flagship product, the ClearPoint Neuro Navigation System, has FDA clearance and is CE-marked. ClearPoint Neuro is engaged with healthcare and research centers in North America, Europe, Asia, and South America. The Company is also partnered with the most innovative pharmaceutical/biotech companies, academic centers, and contract research organizations, providing solutions for direct CNS delivery of therapeutics in pre-clinical studies and clinical trials worldwide. To date, thousands of procedures have been performed and supported by the Companys field-based clinical specialist team, which offers support and services to our customers and partners worldwide. For more information, please visit http://www.clearpointneuro.com.

Forward-Looking Statements

This press release contains forward-looking statements within the context of the federal securities laws, which may include the Companys expectations for the future performance, market, and revenue of its products and services. These forward-looking statements are based on managements current expectations and are subject to the risks inherent in the business, which may cause the Company's actual results to differ materially from those expressed in or implied by forward-looking statements. Particular uncertainties and risks include those relating to: global and political instability, supply chain disruptions, labor shortages, and macroeconomic and inflationary conditions; future revenue from sales of the Companys products and services; the Companys ability to market, commercialize and achieve broader market acceptance for new products and services offered by the Company; the ability of our biologics and drug delivery partners to achieve commercial success, including their use of the Companys products and services in their delivery of therapies; the Companys expectations, projections and estimates regarding expenses, future revenue, capital requirements, and the availability of and the need for additional financing; the Companys ability to obtain additional funding to support its research and development programs; the ability of the Company to manage the growth of its business; the Companys ability to attract and retain its key employees; and risks inherent in the research, development, and regulatory approval of new products. More detailed information on these and additional factors that could affect the Companys actual results are described in the Risk Factors section of the Companys Annual Report on Form 10-K for the year ended December 31, 2023, which has been filed with the Securities and Exchange Commission, and the Companys Quarterly Report on Form 10-Q for the three months ended March 31, 2024, which the Company intends to file with the Securities and Exchange Commission on or before May 15, 2024. The Company does not assume any obligation to update these forward-looking statements.

Excerpt from:
ClearPoint Neuro to Present Novel Research and Exhibit at the 27th American Society of Gene & Cell Therapy Annual ... - GlobeNewswire

San Diego, USA-based clinical-stage cell and gene therapy company Poseida Therapeutics (Nasdaq: PSTX) saw its shares close up more than 20% at $2.92 yesterday, when it revealed a second collaboration with Japanese drug major Astellas Pharma (TYO: 4503).

Astellas wholly-owned Xyphos Bioscience and Poseidahave entered into a research collaboration and license agreement to develop novel convertible CAR programs by combining the innovative cell

This article is accessible to registered users, to continue reading please register for free. A free trial will give you access to exclusive features, interviews, round-ups and commentary from the sharpest minds in the pharmaceutical and biotechnology space for a week. If you are already a registered user pleaselogin. If your trial has come to an end, you cansubscribe here.

Try before you buy

All the news that moves the needle in pharma and biotech. Exclusive features, podcasts, interviews, data analyses and commentary from our global network of life sciences reporters. Receive The Pharma Letter daily news bulletin, free forever.

Become a subscriber

Unfettered access to industry-leading news, commentary and analysis in pharma and biotech. Updates from clinical trials, conferences, M&A, licensing, financing, regulation, patents & legal, executive appointments, commercial strategy and financial results. Daily roundup of key events in pharma and biotech. Monthly in-depth briefings on Boardroom appointments and M&A news. Choose from a cost-effective annual package or a flexible monthly subscription.

Original post:
Astellas and Poseida enter second research collaboration - The Pharma Letter

Yeargin-Allsopp, M. et al. Prevalence of cerebral palsy in 8-year-old children in three areas of the United States in 2002: a multisite collaboration. Pediatrics 121, 547554 (2008).

Article PubMed Google Scholar

Lee, R. W. et al. A diagnostic approach for cerebral palsy in the genomic era. Neuromolecular Med. 16, 821844 (2014).

Article CAS PubMed PubMed Central Google Scholar

Beysen, D. et al. Genetic testing contributes to diagnosis in cerebral palsy: Aicardi-Goutieres syndrome as an example. Front Neurol. 12, 617813 (2021).

Article PubMed PubMed Central Google Scholar

Chang, M. J., Ma, H. I. & Lu, T. H. Estimating the prevalence of cerebral palsy in Taiwan: a comparison of different case definitions. Res. Dev. Disabil. 36C, 207212 (2015).

Article PubMed Google Scholar

MacLennan, A. H., Thompson, S. C. & Gecz, J. Cerebral palsy: causes, pathways, and the role of genetic variants. Am. J. Obstet. Gynecol. 213, 779788 (2015).

Article PubMed Google Scholar

Smithers-Sheedy, H. et al. A special supplement: findings from the Australian Cerebral Palsy Register, birth years 1993 to 2006. Dev. Med. Child Neurol. 58, 510 (2016).

Article PubMed Google Scholar

Sellier, E. et al. Trends in prevalence of cerebral palsy in children born with a birthweight of 2,500 g or over in Europe from 1980 to 1998. Eur. J. Epidemiol. 25, 635642 (2010).

Article PubMed Google Scholar

Gonzalez-Mantilla, P. J. et al. Diagnostic yield of exome sequencing in cerebral palsy and implications for genetic testing guidelines: a systematic review and meta-analysis. JAMA Pediatr. 177, 472478 (2023).

Article PubMed PubMed Central Google Scholar

Moreno-De-Luca, A., Ledbetter, D. H. & Martin, C. L. Genetic insights into the causes and classification of cerebral palsies. Lancet Neurol. 11, 283292 (2012).

Article CAS PubMed PubMed Central Google Scholar

Hemminki, K., Li, X. J., Sundquist, K. & Sundquist, J. High familial risks for cerebral palsy implicate partial heritable aetiology. Paediatr. Perinat. Epidemiol. 21, 235241 (2007).

Article PubMed Google Scholar

Tollanes, M. C., Wilcox, A. J., Lie, R. T. & Moster, D. Familial risk of cerebral palsy: population based cohort study. BMJ 349, g4294 (2014).

Article PubMed PubMed Central Google Scholar

Zhang, G. et al. Genetic associations with gestational duration and spontaneous preterm birth. N. Engl. J. Med. 377, 11561167 (2017).

Article CAS PubMed PubMed Central Google Scholar

Lynex, C. N. et al. Homozygosity for a missense mutation in the 67 kDa isoform of glutamate decarboxylase in a family with autosomal recessive spastic cerebral palsy: parallels with Stiff-Person Syndrome and other movement disorders. BMC Neurol. 4, 20 (2004).

Article PubMed PubMed Central Google Scholar

Lerer, I. et al. Deletion of the ANKRD15 gene at 9p24.3 causes parent-of-origin-dependent inheritance of familial cerebral palsy. Hum. Mol. Genet. 14, 39113920 (2005).

Article CAS PubMed Google Scholar

Abou Jamra, R. et al. Adaptor protein complex 4 deficiency causes severe autosomal-recessive intellectual disability, progressive spastic paraplegia, shy character, and short stature. Am. J. Hum. Genet. 88, 788795 (2011).

Article CAS PubMed PubMed Central Google Scholar

Moreno-De-Luca, A. et al. Adaptor protein complex-4 (AP-4) deficiency causes a novel autosomal recessive cerebral palsy syndrome with microcephaly and intellectual disability. J. Med. Genet. 48, 141144 (2011).

Article CAS PubMed Google Scholar

Verkerk, A. J. et al. Mutation in the AP4M1 gene provides a model for neuroaxonal injury in cerebral palsy. Am. J. Hum. Genet. 85, 4052 (2009).

Article CAS PubMed PubMed Central Google Scholar

Kruer, M. C. et al. Mutations in gamma adducin are associated with inherited cerebral palsy. Ann. Neurol. 74, 805814 (2013).

Article CAS PubMed Google Scholar

Hirata, H. et al. ZC4H2 mutations are associated with arthrogryposis multiplex congenita and intellectual disability through impairment of central and peripheral synaptic plasticity. Am. J. Hum. Genet. 92, 681695 (2013).

Article CAS PubMed PubMed Central Google Scholar

van Eyk, C., MacLennan, S. C. & MacLennan, A. H. All patients with a cerebral palsy diagnosis merit genomic sequencing. JAMA Pediatr. 177, 455456 (2023).

Article PubMed Google Scholar

Liu, J. C. et al. Multi-omics research in sarcopenia: current progress and future prospects. Ageing Res. Rev. 76, 101576 (2022).

Article CAS PubMed Google Scholar

Srivastava, S. et al. Molecular diagnostic yield of exome sequencing and chromosomal microarray in cerebral palsy: a systematic review and meta-analysis. JAMA Neurol. 79, 12871295 (2022).

Article PubMed PubMed Central Google Scholar

Nagy, E. et al. The usefulness of MRI Classification System (MRICS) in a cerebral palsy cohort. Acta Paediatr. 109, 27832788 (2020).

Article PubMed Google Scholar

Himmelmann, K. et al. MRI classification system (MRICS) for children with cerebral palsy: development, reliability, and recommendations. Dev. Med. Child Neurol. 59, 5764 (2017).

Article PubMed Google Scholar

Paneth, N. & Stark, R. I. Cerebral palsy and mental retardation in relation to indicators of perinatal asphyxia. An epidemiologic overview. Am. J. Obstet. Gynecol. 147, 960966 (1983).

Article CAS PubMed Google Scholar

van Rappard, D. F., Boelens, J. J. & Wolf, N. I. Metachromatic leukodystrophy: disease spectrum and approaches for treatment. Best. Pract. Res. Clin. Endocrinol. Metab. 29, 261273 (2015).

Article PubMed Google Scholar

Lerner-Ellis, J. P. et al. Identification of the gene responsible for methylmalonic aciduria and homocystinuria, cblC type. Nat. Genet. 38, 93100 (2006).

Article CAS PubMed Google Scholar

Allewelt, H. et al. Long-term functional outcomes after hematopoietic stem cell transplant for early infantile Krabbe disease. Biol. Blood Marrow Transpl. 24, 22332238 (2018).

Article Google Scholar

Zhong, C. et al. M6A-modified circRBM33 promotes prostate cancer progression via PDHA1-mediated mitochondrial respiration regulation and presents a potential target for ARSI therapy. Int J. Biol. Sci. 19, 15431563 (2023).

Article CAS PubMed PubMed Central Google Scholar

Haussler, M., Hoffmann, G. F. & Wevers, R. A. l-DOPA and selegiline for tyrosine hydroxylase deficiency. J. Pediatr. 138, 451452 (2001).

Article CAS PubMed Google Scholar

Huppke, P. et al. Activating de novo mutations in NFE2L2 encoding NRF2 cause a multisystem disorder. Nat. Commun. 8, 818 (2017).

Article PubMed PubMed Central Google Scholar

Friedman, J., Hyland, K., Blau, N. & MacCollin, M. DOPA-responsive hypersomnia and mixed movement disorder due to sepiapterin reductase deficiency. Neurology 67, 20322035 (2006).

Article PubMed Google Scholar

Moreno-De-Luca, A. et al. Molecular diagnostic yield of exome sequencing in patients with cerebral palsy. JAMA 325, 467475 (2021).

Article CAS PubMed PubMed Central Google Scholar

Ioannidis, N. M. et al. REVEL: an ensemble method for predicting the pathogenicity of rare missense variants. Am. J. Hum. Genet. 99, 877885 (2016).

Article CAS PubMed PubMed Central Google Scholar

Ke, H. N. et al. Landscape of pathogenic mutations in premature ovarian insufficiency. Nat. Med. 29, 483492 (2023).

Article CAS PubMed PubMed Central Google Scholar

McMichael, G. et al. Whole-exome sequencing points to considerable genetic heterogeneity of cerebral palsy. Mol. Psychiatry 20, 176182 (2015).

Article CAS PubMed Google Scholar

Takezawa, Y. et al. Genomic analysis identifies masqueraders of full-term cerebral palsy. Ann. Clin. Transl. Neurol. 5, 538551 (2018).

Article CAS PubMed PubMed Central Google Scholar

Matthews, A. M. et al. Atypical cerebral palsy: genomics analysis enables precision medicine. Genet. Med. 21, 16211628 (2019).

Article CAS PubMed Google Scholar

Bamshad, M. J., Nickerson, D. A. & Chong, J. X. Mendelian gene discovery: fast and furious with no end in sight. Am. J. Hum. Genet. 105, 448455 (2019).

Article CAS PubMed PubMed Central Google Scholar

van Eyk, C. L. et al. Analysis of 182 cerebral palsy transcriptomes points to dysregulation of trophic signalling pathways and overlap with autism. Transl. Psychiatry 8, 88 (2018).

Article PubMed PubMed Central Google Scholar

van Eyk, C. L. et al. Yield of clinically reportable genetic variants in unselected cerebral palsy by whole genome sequencing. NPJ Genom. Med. 6, 74 (2021).

Article PubMed PubMed Central Google Scholar

MacLennan, A. A template for defining a causal relation between acute intrapartum events and cerebral palsy: international consensus statement. BMJ 319, 10541059 (1999).

Article CAS PubMed PubMed Central Google Scholar

OBrien, J. R., Usher, R. H. & Maughan, G. B. Causes of birth asphyxia and trauma. Can. Med. Assoc. J. 94, 10771085 (1966).

PubMed PubMed Central Google Scholar

Windle, W. F. Neurological and psychological deficits from asphyxia neonatorum. Public Health Rep. (1896) 72, 646650 (1957).

Article CAS PubMed Google Scholar

Phelan, J. P., Martin, G. I. & Korst, L. M. Birth asphyxia and cerebral palsy. Clin. Perinatol. 32, 6176 (2005).

Article PubMed Google Scholar

Sartwelle, T. P., Johnston, J. C. & Arda, B. A half century of electronic fetal monitoring and bioethics: silence speaks louder than words. Matern. Health Neonatol. Perinatol. 3, 21 (2017).

Article PubMed PubMed Central Google Scholar

Zarrei, M. et al. De novo and rare inherited copy-number variations in the hemiplegic form of cerebral palsy. Genet. Med. 20, 172180 (2018).

Article PubMed Google Scholar

Han, V. X., Patel, S., Jones, H. F. & Dale, R. C. Maternal immune activation and neuroinflammation in human neurodevelopmental disorders. Nat. Rev. Neurol. 17, 564579 (2021).

Article PubMed Google Scholar

Jung, E. et al. Clinical chorioamnionitis at term: definition, pathogenesis, microbiology, diagnosis, and treatment. Am. J. Obstet. Gynecol. 230, S807S840 (2024).

Article PubMed Google Scholar

Prieto-Villalobos, J. et al. Astroglial hemichannels and pannexons: the hidden link between maternal inflammation and neurological disorders. Int. J. Mol. Sci. 22, 9503 (2021).

Article CAS PubMed PubMed Central Google Scholar

Excerpt from:
Exome sequencing reveals genetic heterogeneity and clinically actionable findings in children with cerebral palsy - Nature.com

WATCH TIME: 6 minutes

"Geographic diversity is incredibly important in Parkinson research. Tasso's device helps reach patients in remote areas where access to phlebotomists may be limited."

PD GENEration: Mapping the Future of Parkinsons Disease,an international genetics study led by the Parkinsons Foundation, tests for mutations among select, clinically relevant genes to accelerate research, advance treatments, and improve care for patients with Parkinson disease (PD). The goal of the study is to make genetic testing accessible to patients with PD, empower those with the disease and their clinicians to know their genetic status, and identify clinical trials in which they might enroll. PD GENEration returns genetic findings to all participants through a genetic counseling session, bridging the knowledge gap between patients and clinicians to accelerate research collectively.

In recent news, the foundation announced a partnership with Tasso that would leverage the companys patient-centric, end-to-end sample collection and logistics platform for PD GENEration.1 Patients in the study will receive a kit containing a Tasso device that will collect a small blood sample in the comfort and convenience of their home. Online proctors will help guide participants through the collection process and provide support for a successful collection. After collection, patients can ship their sample in a pre-paid box to a lab for research analysis. Overall, the goal of this research is to screen the collected samples for mutations among select, clinically relevant PD genes.

James Beck, PhD, senior vice president and chief scientific officer of the Parkinsons Foundation, and Ben Casavant, PhD, CEO and cofounder of Tasso, recently sat down with NeurologyLive in an interview to discuss how the blood collection device can simplify the process of collecting a sample of blood compared with traditional methods. The duo also talked about the role of genetic analysis in Parkinson research, and how Tasso is contributing to it. Additionally, Beck and Casavant talked about the importance of geographic diversity in the context of Parkinson research and patient access to testing.

Click here to learn more about PD GENEration.

Excerpt from:
Using a Blood Collection Device to Advance Genetic Research in Parkinson Disease: James Beck, PhD & Ben ... - Neurology Live

A high-risk cancer genetics program at Erlanger is helping improve cancer treatment and screenings for patients in our community.

Actress Olivia Munn says a breast cancer risk assessment helped save her life. She announced last year that she had been diagnosed with Luminal B Cancer, which is an aggressive, fast-moving cancer.

A similar program is available at Erlanger.

McKenzie Smartt, Erlanger, NP-C says, Our cancer genetics program we can do genetic testing to test your DNA to see if you have any hereditary conditions that were passed down from your family that could possibly increase your risk for cancer.

Smartt says patients can also enroll in their high-risk program.

McKenzie Smartt says, Where if you do not have anything genetic you can still have an increased risk for cancer based off family history. So, we can manage the increased risk as well.

Smartt says everyones risk is different. While this program is for everyone, it is geared toward women.

McKenzie Smartt says Specifically, women over the age of 25. The guidelines now suggest women over 25 should have a risk assessment.

A lot of that has to do with the fact that 1 in 8 women will be diagnosed with breast cancer in their lifetime.

Smartt says they also are concerned about other cancers like colon cancer and prostate cancer.

McKenzie Smartt says We want to be proactive in the community and be able to identify those who are at risk, catch cancer early so we can begin treatment as quickly as possible.

For more details, click here.

Visit link:
EYE ON HEALTH: High risk and genetic testing being offered at local hospital - Local 3 News

GREENFIELD It took nearly 35 years, but the human remains found just off Route 78 in Warwick in 1989 have been identified as belonging to Constance (Holminski) Bassignani, who was 65 years old at the time of her murder.

The Northwestern District Attorneys Office held a press conference Thursday morning to announce that the victims identity was learned about eight months after her DNA was submitted to Othram, a Texas corporation that specializes in using forensic genetic genealogy to resolve unsolved murders, disappearances and identification of unidentified decedents or homicide victims.

According to the DAs office, Bassignani was born in Hawaii in 1924 and was living with her second husband, William Bassignani, in Woonsocket, Rhode Island, at the time of her death. William reportedly told family that she had moved back to Hawaii, though the DAs office stated investigators found no evidence of this. William, who died in 1993, is considered a person of interest in the case.

The State Police Detective Unit attached to DAs office collaborated with the State Police Crime Laboratory to submit the genetic material to Othram. This led to living potential relatives, who submitted their own DNA and confirmed the victims identity.

Were all very appreciative of the dedication, the hard work and the perseverance in this case, Northwestern District Attorney David Sullivan said. Seeking justice for the unknown victim has been their driving force from Day 1.

Bassignanis body was found on June 24, 1989, by a passing motorist. Her remains were found 10 to 20 feet off the roadway of Route 78, in a lightly wooded area near a small gravel pit about 1 miles south of the New Hampshire state line and about the same distance north of the entrance to Mount Grace State Forest. Sullivan said local and State Police responded to the scene, documenting evidence and collecting the remains.

First Assistant District Attorney Steven Gagne explained that identifying the victim is the first step in any homicide investigation.

From there, investigators can determine who the victims circle of family, friends and co-workers were and attempt to retrace their last known steps and contacts, he said. In this case, investigators were hampered from the start in their efforts to solve this homicide without an identification of the victim.

Article continues after...

Gagne said investigators have learned Bassignani got married and divorced in the 1940s before marrying William in 1945. Authorities have tracked down and spoken with the three grandchildren born to a son that has died, as well as a daughter who lives on the West Coast. Gagne said the DAs office was assisted by law enforcement in Washington state and in Hawaii.

So this investigation, which is now reinvigorated, has literally spanned half the globe, he said.

Gagne explained Bassignani and her husband lived in an apartment in Woonsocket, Rhode Island, approximately 80 miles from Warwick. The couple had previously lived in Franklin, Massachusetts. Sullivan said there is no known connection between the couple and Warwick, and it is unknown where the murder took place.

According to the DAs office, the victim was last seen alive on Memorial Day weekend in 1989. Gagne said William reportedly told relatives that his wife had decided to move back to Hawaii and that they would not be seeing or hearing from her again.

Gagne said he hopes this news provides some closure to the victims family, shedding some light on what may have been a looming cloud of doubt surrounding her disappearance for decades.

Retired Warwick Police Chief Brian Peters, who was at the departments helm in 1989, said it is refreshing to get some answers.

Its a big relief, he said. [We] never had anything like this happen in town, and hopefully never will [again].

Paul Marguet, the State Polices lead investigator on this case, said the victims grandchildren initially did not believe authorities, having long believed their grandfathers story about his wifes abrupt disappearance. But, he said, their DNA was used to confirm the truth.

Michael Vogen, an Othram representative, appeared via Zoom during Thursdays press conference and lauded the DAs office and the State Police before briefly speaking about his companys mission.

We were purpose-built to do just this, he said. Thats to generate human ID from forensic evidence.

Gagne previously told the Greenfield Recorder the decision was made to reach out to Othram due to the companys success in identifying the so-called Granby Girl as Patricia Ann Tucker, a 28-year-old woman found shot to death in 1978, but who went unidentified until a little over a year ago. Gerald Coleman, Tuckers husband when she died, is a person of interest in that murder. He died in state prison in 1996, and prosecutors say he never reported his wife as missing.

Othram also helped identify the Lady of the Dunes a 37-year-old woman found murdered in Provincetown on July 26, 1974 as Ruth Marie Terry. Her now-deceased husband, Guy Muldavin, was officially named as the killer in August 2023.

David Mittelman, founder and CEO of Othram, previously told the Recorder that his company tests DNA based on hundreds of thousands of markers, whereas the FBIs Combined DNA Index System (CODIS) uses 20. He also said victims are often not in CODIS because it was designed about 30 years ago to track the repeat offenses of known criminals. But Othram, which employs 60 people, can work from evidence generally considered unusable because it is too old or too degraded.

Gagne mentioned authorities hope to bring renewed attention to the Warwick case and trigger some new leads that have a domino effect that ends with additional answers. Anyone with information that might be helpful in this case is encouraged to call the State Police Detective Unit attached to the Northwestern District Attorneys Office at 413-512-5361. Messages can also be submitted anonymously through northwesternda.org.

Its like were trying to piece together an ancient puzzle here, Gagne told reporters, but any small piece would certainly help.

Reach Domenic Poli at: dpoli@recorder.com or 413-930-4120.

Read the original here:
1989 homicide victim found in Warwick ID'd through genetic testing, but some mysteries remain - The Recorder

WASHINGTON, May 2, 2024 /PRNewswire/ -- One of the biggest scams targeting Medicare beneficiaries in the last decade has been the genetic testing scam. Recently, the Senior Medicare Patrol (SMP) has seen an increase in genetic testing complaints. Across the nation, genetic testing company representatives are offering "free" genetic tests, also referred to as DNA screenings, cancer screenings, and hereditary testing, in exchange for the beneficiaries' Medicare numbers.

Beneficiaries are getting calls about genetic tests claiming that the results will help them avoid or detect diseases like cancer or Alzheimer's. The SMP has also received reports of genetic testing claims on Medicare statements when the beneficiaries never received any contact about genetic testing. It is incredibly important to review your Medicare statements and report these claims.

The genetic testing scam can be dangerous. "Scammers can steal people's medical identity and falsely bill Medicare (around $10,000 a claim), draining the Medicare program. Additionally, tests ordered under these circumstances could lead to confusion and inaccurate medical records," said Nicole Liebau, SMP Resource Center director.

The SMP recommends that Medicare beneficiaries:

The Senior Medicare Patrol (SMP) is ready to provide you with the information you need to PROTECT yourself from Medicare fraud, errors, and abuse; DETECT potential fraud, errors, and abuse; and REPORT your concerns. SMPs help educate and empower Medicare beneficiaries in the fight against health care fraud. Your SMP can help you with your questions, concerns, or complaints about potential fraud and abuse issues. It also provides information and educational presentations. To locate the local Senior Medicare Patrol, contact 1-877-808-2468 or go to http://www.smpresource.org.

SMP Resource Center Nicole Liebau 319-284-0702 [emailprotected]

SOURCE SMP Resource Center

Read this article:
Double Helix Deception: The Return of Genetic Testing Scams in Medicare Fraud - PR Newswire

Finally, the moment arrived for my BRCA test a simple blood draw, almost anticlimactic for something that had the potential to be life-altering and life-saving. But how did I get here?

This moment had been something Id been building up in my head since 2017 when I saw The Bold Type and Jane Sloans personal journey as she navigated her mothers battle with breast cancer and her own subsequent decision to undergo genetic testing to look for BRCA genes. Now, here I was at my very own appointment in 2024. Sure, maybe it took me seven years but I was finally confronting, and struggling to manage, the overwhelming mix of anxiety, fear, grief, and empowerment I felt (but at least my therapist is fantastic). I was facing my own mortality squarely in the face and saying bring it.

So here I was, at a large hospital, as ready as I could be for my hour-long session where my family tree, personal history, and risks would be reviewed and evaluated. I was immediately struck by the warmth and empathy of my genetic counselor, whose name was Emily. She was kind and welcoming to both me and my partner, and never said any of the awkward veiled homophobia I usually hear in medical offices like is this your friend or worse, is this your sister. She was able to delicately balance the clinical aspects of genetics with the emotional weight of familial cancer history.

Most of my known family history and risk is on one half of my family, and the cancers that show up in my family tree can all be linked genetically. The counselor took all of this anecdotal information and plugged it into a mathematical formula. She calculated the risk that I would have a genetic mutation that is known to cause cancer. Even with only one half of my family history known to me, Emily confirmed my risk level was considered high enough to make me a strong candidate for testing.

It was strange hearing that out loud. I had already done my own research and knew that was likely to be the outcome, but having an expert confirm that I had a high risk level was both scary and exciting. I wanted to get the test, even if doing so seemed like asking a psychic when I was going to die.

Emily gave me information regarding my insurance coverage and financial support available through the genetic testing lab itself (not the hospital where I was doing the testing, but the organization that would actually analyze my genes). Fortunately, because of fairly recent health laws that require insurance to cover BRCA testing for high risk patients, my insurance would cover the test based on the fact that I was confirmed to be a good candidate.

Next it was time to consider how extensive I wanted the test to be, which required thinking about some of the most complex parts of the entire process: mastectomy, hormone therapy, hysterectomy, and more.

The genetic counselor was again fantastic and made sure that I didnt boil over like an out-of-control pot of water. She told me that the recommendations for medical intervention can vary based on the specific gene combination that is found. For example, a mastectomy might be a recommendation for a really high risk gene profile, but monitoring and hormone treatment might be the recommendation for a more moderate risk gene profile.

The question of whether or not to get a hysterectomy or mastectomy or even do hormone therapy is inextricably tied to notions of gender and what that means, personally and societally. I was first introduced to those conversations ten years ago, when Angelina Jolie revealed that she had gotten a mastectomy because of her own BRCA mutation. At the time it terrified me. It sounded like such an intense surgery and it was impossible for me, at the time, to separate breasts from femininity. The media confirmed this with countless op-eds at the time arguing whether or not Angelina Jolie was still a woman.

Of course, I no longer believe that breasts are necessary to be a woman. Trans women are women, even if they dont take hormones or get top surgery.I could still be a woman after a mastectomy. Tig Notaro, who I adore, went on stage shirtless r in her comedy special Boyish Girl Interrupted after a mastectomy because of breast cancer, and she identifies as a woman and a lesbian. Id also spent time as the primary caretaker for someone recovering from top surgery and had first hand knowledge about the bandages, drains, scars, and all of the details in between. Top surgery and a mastectomy arent exactly the same, but the surgeries have a lot in common, and my queer experience gave me a unique perspective to bear witness to the ways in which breast removal could be a joyful experience and not solely something done out of the fear and pain that comes with cancer. It helped me understand the surgery from a practical perspective, rather than the fear and tragedy that often accompanies a mastectomy in the case of cancer.

Hysterectomy and hormones are a little harder to comprehend, because they are not as visible and public. I also have endometriosis and the lesser-known adenomyosis, both of which mean that the lining of the uterus grows where they are not supposed to, meaning it is wildly painful and extraordinarily torturous to have a period. For me, a hysterectomy would address these conditions as well, making it feel easier to consider such a big surgery. Pair all of this with the BRCA statistics: A BRCA mutation can cause up to a 72% lifetime risk of developing cancer and a 44% lifetime risk of developing ovarian cancer. For those with BRCA who do develop cancer, it is much more likely to be aggressive and more likely to be deadly. I had already decided if I had high genetic risk, I wanted to take an aggressive approach to fighting and preventing it.

In fact, my coming to this specific hospital was the result of a lot of research to confirm that things like hysterectomy would be an option. Navigating the healthcare system as a disabled LGBTQ individual in a post-Roe and post-Trump America is far from straightforward, and I knew that it was possible that religious hospitals and state laws could affect what treatment options are available to me. Having received affirming care at One Medical, known for its LGBTQ+-friendly environment and inclusive practices, I was taken aback when I was first referred to Providence hospitals. Despite Providences outward portrayal of inclusivity, deeper research revealed instances of discriminatory practices, such as denying transgender individuals necessary medical procedures or restricting access to birth control and abortion based on religious doctrine. Choosing a hospital system became a key part of my process and I had to do extra work to ensure comprehensive gynecologic care options and avoid discriminatory practices. I didnt want to risk being denied all of the possible treatment options.

Now, sitting with Emily, I felt confident that I made the right decision. I wouldnt have to worry about artificial restrictions on my healthcare. Instead I could just consider the range of genetic test options knowing that even if the test revealed a high genetic cancer risk, this hospital had a team who would give me all of the options.

Finally, it was time to decide how many genes I actually wanted to have tested. They could perform a targeted test focusing on specific cancer-linked mutations or a comprehensive panel. Emily noted that one reason to choose more targeted testing was because going with the full panel increased the odds of finding an unknown genetic mutation and that this could cause anxiety. Scientific understanding of genes and cancer is still evolving, and some genetic variations have been identified, but not studied enough to conclusively say whether or not they increase the risk for cancer. However, she assured me that the testing company would provide ongoing updates, ensuring I remained informed as the science developed. I decided to go with the full, comprehensive panel of eighty eight genes, choosing to confront the unknown with as much data as possible. Id prefer to have all available information that science could offer me.

(Plus, the cost for the test would be the same whether I tested all eighty eight genes or a much smaller number, and I love a good bargain. Eighty-eight genes for the price of two, yes please!)

It feels like after all of the build up the actual genetic test should take place around a campfire with chanting and handholding, but the reality was anticlimactic and impersonal. If anything, the test itself felt really awkward. The nurse struggled to take my blood even though I am usually told by medical professionals that I have good veins. It felt like some part of my body was determined to express the anxiety I still felt by physically fighting back against the test. Finally after several minutes of poking at me the blood began to flow, they collected two small tubes, packaged it in a box to send to the lab, and I was finished.

As I left the appointment, I carried a sense of relief tinged with apprehension, knowing that the next few weeks would surely make my anxiety boil over.

But what convinced me to do the test in the first place remains true: if I have the genetic mutations that increase my risk for multiple types of cancer, especially breast cancer, theyre already there whether or not I get the test.

Now, as I await the verdict of eighty-eight potential cancer-causing genes, I am buoyed by the legacy of LGBTQ+ people who came before and whose courage and honesty helped bring awareness, empathy, and less stigmatization to breast cancer and the gender-based shame that can come with it. There is Wanda Sykes, who opted for a double mastectomy after finding what she called stage 0 cancer because she wanted to reduce her chance of it spreading as much as possible. Melissa Etheridge, who destigmatized the connection between breast cancer and femininity by performing bald at the GRAMMY Awards shortly after completing chemotherapy. Robin Roberts and her partner Amber Laig,n who both have had breast cancer and shared their experiences via Robins platform as a host on Good Morning America. Angelina Jolie, and her pivotal role in bringing BRCA genes and treatment options into the national spotlight. Audre Lorde, who published The Cancer Journals and detailed her own struggles with and views of post-cancer femininity.

I also still think of Tig Notaro, who in the middle of a stand-up comedy special being recorded for broadcast boldly unbuttoned her shirt, revealing her post-mastectomy chest. She did the remaining half of her stand-up set like this, forcing viewers to confront their own discomfort with cancer, mortality, gender, and health all in one subtle but significant move.

Tig stands on a stage for 20 minutes, literally in a spotlight and on camera, shamelessly showing her nipple-less, slightly concave chest with obvious red surgery scars. The first time I watched it I felt uncomfortable, being forced to face medical and gender stigmas at the same time while feeling amazed at Tigs boldness. It felt like I was watching something public that was supposed to be kept hidden and private and it was inspiring to see that see the mystery, and with it the stigma, stripped away. Its what I aim to do with this series in some small, similar way. I think perhaps if the worst case scenario for my health and my gender presentation is that I have something in common with Tig Notaro then maybe thats not so bad.

What to Expect Whenis a series from Katie Reilly shedding light on cancer and the intersection of genetics, identity, and health.

Join AF+!

Read the original here:
What to Expect While Awaiting the Results of a Life Changing Test - http://www.autostraddle.com