Molecular Therapy-Methods & Clinical Development最新文献_第6页

Intrathecal or intravenous AAV9-IDUA/RGX-111 at minimal effective dose prevents cardiac, skeletal and neurologic manifestations of murine MPS I. 鞘内或静脉注射最小有效剂量的AAV9-IDUA/RGX-111可预防小鼠MPS I的心脏、骨骼和神经表现。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-04 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101369

Lalitha R Belur, Avery K Huber, Hillary Mantone, Mason Robertson, Miles C Smith, Andrea D Karlen, Kelley F Kitto, Li Ou, Chester B Whitley, Elizabeth Braunlin, Justin Furcich, Troy C Lund, Davis Seelig, Carolyn A Fairbanks, Nicholas Buss, Kwi Hye Kim, R Scott McIvor

{"title":"Intrathecal or intravenous AAV9-IDUA/RGX-111 at minimal effective dose prevents cardiac, skeletal and neurologic manifestations of murine MPS I.","authors":"Lalitha R Belur, Avery K Huber, Hillary Mantone, Mason Robertson, Miles C Smith, Andrea D Karlen, Kelley F Kitto, Li Ou, Chester B Whitley, Elizabeth Braunlin, Justin Furcich, Troy C Lund, Davis Seelig, Carolyn A Fairbanks, Nicholas Buss, Kwi Hye Kim, R Scott McIvor","doi":"10.1016/j.omtm.2024.101369","DOIUrl":"10.1016/j.omtm.2024.101369","url":null,"abstract":"Mucopolysaccharidosis type I (MPS I) is a rare metabolic disorder caused by deficiency of α-L-iduronidase (IDUA), resulting in glycosaminoglycan (GAG) accumulation and multisystemic disease. Current treatments include hematopoietic stem cell transplantation and enzyme replacement therapy, but these do not address all manifestations of the disease. We infused MPS I mice with an adeno-associated virus 9 (AAV9)-IDUA vector (RGX-111) at doses from 107 to 1010 vector genomes (vg) via intrathecal (IT), intravenous (IV), and intrathecal+intravenous (IT+IV) routes of administration. In mice administered doses ≤109 vg IT or ≤108 vg IV, there was no therapeutic benefit, while in mice administered 109 vg IV, there was a variable increase in IDUA activity with inconclusive neurocognitive and cardiac assessments. However, at the 1010 vg dose, we observed substantial metabolic correction, with restored IDUA levels and normalized tissue GAGs for all treatment groups. Aortic insufficiency was mostly normalized, neurologic deficit was prevented, and microcomputed tomography (micro-CT) analysis showed normalization of skeletal parameters. Histologic analysis showed minimal GAG storage and lysosomal pathology. We thus report a minimal effective dose of 1010 vg (5 × 1011 per kg) RGX-111 for IV and IT routes of administration in MPS I mice, which prevented neurocognitive deficit, cardiac insufficiency, and skeletal manifestations, as a model for genetic therapy of human MPS I.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101369"},"PeriodicalIF":4.6,"publicationDate":"2024-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11646787/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142840384","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Comprehensive analysis of off-target and on-target effects resulting from liver-directed CRISPR-Cas9-mediated gene targeting with AAV vectors. 肝靶向crispr - cas9介导的AAV载体基因靶向的脱靶和靶标效应综合分析

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-04 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101365

Kshitiz Singh, Raffaele Fronza, Hanneke Evens, Marinee K Chuah, Thierry VandenDriessche

{"title":"Comprehensive analysis of off-target and on-target effects resulting from liver-directed CRISPR-Cas9-mediated gene targeting with AAV vectors.","authors":"Kshitiz Singh, Raffaele Fronza, Hanneke Evens, Marinee K Chuah, Thierry VandenDriessche","doi":"10.1016/j.omtm.2024.101365","DOIUrl":"10.1016/j.omtm.2024.101365","url":null,"abstract":"Comprehensive genome-wide studies are needed to assess the consequences of adeno-associated virus (AAV) vector-mediated gene editing. We evaluated CRISPR-Cas-mediated on-target and off-target effects and examined the integration of the AAV vectors employed to deliver the CRISPR-Cas components to neonatal mice livers. The guide RNA (gRNA) was specifically designed to target the factor IX gene (F9). On-target and off-target insertions/deletions were examined by whole-genome sequencing (WGS). Efficient F9-targeting (36.45% ± 18.29%) was apparent, whereas off-target events were rare or below the WGS detection limit since only one single putative insertion was detected out of 118 reads, based on >100 computationally predicted off-target sites. AAV integrations were identified by WGS and shearing extension primer tag selection ligation-mediated PCR (S-EPTS/LM-PCR) and occurred preferentially in CRISPR-Cas9-induced double-strand DNA breaks in the F9 locus. In contrast, AAV integrations outside F9 were not in proximity to any of ∼5,000 putative computationally predicted off-target sites (median distance of 70 kb). Moreover, without relying on such off-target prediction algorithms, analysis of DNA sequences close to AAV integrations outside the F9 locus revealed no homology to the F9-specific gRNA. This study supports the use of S-EPTS/LM-PCR for direct in vivo comprehensive, sensitive, and unbiased off-target analysis.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101365"},"PeriodicalIF":4.6,"publicationDate":"2024-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11626537/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142803323","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Repeated dosing of AAV-mediated liver gene therapy in juvenile rat and mouse models of Crigler-Najjar syndrome type I. aav介导的肝基因治疗在ⅰ型Crigler-Najjar综合征幼鼠和小鼠模型中的应用

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-10-28 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101363

Xiaoxia Shi, Giulia Bortolussi, Fanny Collaud, Pierre-Romain Le Brun, Lysbeth Ten Bloemendaal, Nicolas Guerchet, Dirk Rudi de Waart, Pauline Sellier, Suzanne Duijst, Philippe Veron, Federico Mingozzi, Takashi Kei Kishimoto, Giuseppe Ronzitti, Piter Bosma, Andrés F Muro

{"title":"Repeated dosing of AAV-mediated liver gene therapy in juvenile rat and mouse models of Crigler-Najjar syndrome type I.","authors":"Xiaoxia Shi, Giulia Bortolussi, Fanny Collaud, Pierre-Romain Le Brun, Lysbeth Ten Bloemendaal, Nicolas Guerchet, Dirk Rudi de Waart, Pauline Sellier, Suzanne Duijst, Philippe Veron, Federico Mingozzi, Takashi Kei Kishimoto, Giuseppe Ronzitti, Piter Bosma, Andrés F Muro","doi":"10.1016/j.omtm.2024.101363","DOIUrl":"10.1016/j.omtm.2024.101363","url":null,"abstract":"Crigler-Najjar syndrome is an ultra-rare monogenic recessive liver disease caused by UGT1A1 gene mutations. Complete UGT1A1 deficiency results in severe unconjugated hyperbilirubinemia in newborns that, if not treated, may lead to brain damage and death. Treatment is based on intensive phototherapy, but its efficacy decreases with age, rendering liver transplantation the only curative option. Adeno-associated virus (AAV)-mediated gene therapy has shown long-term correction in adult patients, but loss of viral DNA and therapeutic efficacy are expected in younger patients associated with liver growth. Effective vector re-administration is hindered by anti-AAV neutralizing antibodies generated during the first administration. Here, we investigated AAV vector re-administration by modulating the immune response with rapamycin-loaded nanoparticles (ImmTOR) in Gunn rats (Ugt1a -/- ) and Ugt1a -/- mice. We administered a liver-specific AAV8 vector expressing a codon-optimized hUGT1A1 cDNA (1.0E11 vg/kg) in P25-P28 mutant animals and, upon loss of efficacy after 3 to 5 weeks, a higher second dose (1.0E12 or 5.0E12 vg/kg) was given. ImmTOR co-administration reduced anti-AAV neutralizing antibodies and immunoglobulin Gs generation in male animals of both models allowing effective re-dosing, underscored by a significant and long-term decrease in plasma bilirubin, although efficacy was affected by low-titer residual anti-AAV antibodies suggesting that re-administration in patients may require combination with other methods.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101363"},"PeriodicalIF":4.6,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11607602/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142774903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Ocular toxicity, distribution, and shedding of intravitreal AAV-eqIL-10 in horses. 视网膜内 AAV-eqIL-10 对马眼部的毒性、分布和脱落。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-10-28 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101360

Kim Young, Tomoko Hasegawa, Naveen Vridhachalam, Nichol Henderson, Jacklyn H Salmon, Trace F McCall, Matthew L Hirsch, Brian C Gilger

{"title":"Ocular toxicity, distribution, and shedding of intravitreal AAV-eqIL-10 in horses.","authors":"Kim Young, Tomoko Hasegawa, Naveen Vridhachalam, Nichol Henderson, Jacklyn H Salmon, Trace F McCall, Matthew L Hirsch, Brian C Gilger","doi":"10.1016/j.omtm.2024.101360","DOIUrl":"10.1016/j.omtm.2024.101360","url":null,"abstract":"Non-infectious uveitis (NIU) is a painful recurrent disease affecting 2%-5% of horses. Current treatments require frequent administration with associated adverse events. In a previous study, intravitreal (IVT) adeno-associated virus (AAV) harboring equine interleukin-10 (eqIL-10) cDNA inhibited experimental uveitis in rats. The goal of this study was to evaluate the ocular tolerability, vector genome (vg) distribution, and vector shedding following an IVT injection of AAV8-eqIL-10 in normal horses with the hypothesis that it would be well tolerated in a dose-dependent manner in horses. Injections were well tolerated with mild transient signs of ocular inflammation; however, horses receiving the highest dose developed keratic precipitates. The vgs were not detected in the tears 3 days after injection, or in urine or feces at any time. Aqueous and vitreous humor eqIL-10 levels increased to higher than 1.5 ng/mL, more than 20 times higher than reported effective endogenous and induced levels. The vgs were detected in ocular tissues, and systemic distribution was identified only in the liver and kidney. No systemic effects were identified 86 days after dosing with IVT AAV-eqIL-10. Further investigation of lower doses of IVT AAV8-eqIL-10 therapy is an important next step toward a safe and effective single-dose treatment of equine uveitis with broader implications for treating NIU in humans.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101360"},"PeriodicalIF":4.6,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11656199/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142866292","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Off-the-shelf allogeneic natural killer cells for the treatment of COVID-19. 用于治疗COVID-19的现成同种异体自然杀伤细胞。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-10-28 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101361

Winnie L Liu, Eleftheria Kampouri, John K Bui, Mandeep K Sekhon, Almudena Tercero, Dan Finlay, Liya H Asghedom, Gladys R Romasanta, Natalie T Rice, Fatima Ranjbaran, Carrie Stoltzman, Jody Cook, Joe Blake, Colleen S Delaney, Joshua A Hill

{"title":"Off-the-shelf allogeneic natural killer cells for the treatment of COVID-19.","authors":"Winnie L Liu, Eleftheria Kampouri, John K Bui, Mandeep K Sekhon, Almudena Tercero, Dan Finlay, Liya H Asghedom, Gladys R Romasanta, Natalie T Rice, Fatima Ranjbaran, Carrie Stoltzman, Jody Cook, Joe Blake, Colleen S Delaney, Joshua A Hill","doi":"10.1016/j.omtm.2024.101361","DOIUrl":"10.1016/j.omtm.2024.101361","url":null,"abstract":"Low levels and function of natural killer (NK) cells are associated with increased coronavirus disease 2019 (COVID-19) severity. NK cell immunotherapy may improve immune function to reduce infection severity. We conducted a first-in-human, open-label, phase 1, dose-escalating (100 × 106, 300 × 106, or 900 × 106 cells) study of a single dose of DVX201, a cord-blood-derived allogeneic NK cell therapy, in hospitalized patients with COVID-19. Participants were followed for 28 days. The maximum allowed steroid dose for eligibility was up to 0.5 mg/kg prednisone (or equivalent) daily. We enrolled nine participants, 3 per dose level. Eight participants had ≥1 comorbidity associated with increased COVID-19 severity, three of whom had a hematologic malignancy. Infusions were well tolerated, with no treatment-related adverse events. There was no evidence of inflammatory complications related to infusions. Peripheral blood NK cells generally increased after infusion, peaking by day 7. The median time from infusion to discharge was 2 days (range: 1-13). Two patients (both with acute lymphoblastic leukemia) were readmitted with recurrent COVID-19. This trial demonstrates the safety of allogeneic NK cell immunotherapy as a potential antiviral. Larger controlled trials are needed to establish efficacy.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101361"},"PeriodicalIF":4.6,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11609367/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142774901","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Design and validation of cell-based potency assays for frataxin supplementation treatments. 细胞效价法的设计与验证。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-27 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101347

Shibani Mukherjee, Larisa Pereboeva, Daniel Fil, Achisha Saikia, Jeon Lee, Jixue Li, M Grazia Cotticelli, Elisabetta Soragni, Robert B Wilson, Marek Napierala, Jill S Napierala

{"title":"Design and validation of cell-based potency assays for frataxin supplementation treatments.","authors":"Shibani Mukherjee, Larisa Pereboeva, Daniel Fil, Achisha Saikia, Jeon Lee, Jixue Li, M Grazia Cotticelli, Elisabetta Soragni, Robert B Wilson, Marek Napierala, Jill S Napierala","doi":"10.1016/j.omtm.2024.101347","DOIUrl":"10.1016/j.omtm.2024.101347","url":null,"abstract":"Friedreich's ataxia (FRDA) is a multisystem, autosomal recessive disorder caused by mutations in the frataxin (FXN) gene. As FRDA is considered an FXN deficiency disorder, numerous therapeutic approaches in development or clinical trials aim to supplement FXN or restore endogenous FXN expression. These include gene therapy, protein supplementation, genome editing or upregulation of FXN transcription. To evaluate efficacy of these therapies, potency assays capable of quantitative determination of FXN biological activity are needed. Herein, we evaluate the suitability of mouse embryonic fibroblasts derived from Fxn G127V knockin mice (MUT MEFs) as a candidate for cell-based potency assays. We demonstrate that these cells, when immortalized, continue to express minute amounts of Fxn and exhibit a broad range of phenotypes that result from severe Fxn deficiency. Exogenous FXN supplementation reverses these phenotypes. Thus, immortalized MUT MEFs are an excellent tool for developing potency assays to validate novel FRDA therapies. Care needs to be exercised while utilizing these cell lines, as extended passaging results in molecular changes that spontaneously reverse FRDA-like phenotypes without increasing Fxn expression. Based on transcriptome analyses, we identified the Warburg effect as the mechanism allowing cells expressing a minimal level of Fxn to thrive under standard cell culture conditions.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101347"},"PeriodicalIF":4.6,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11735916/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143016389","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Adeno-associated virus serotype 9 antibodies in neonates and young children: Seroprevalence and kinetics. 腺相关病毒血清型9抗体在新生儿和幼儿：血清阳性率和动力学。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-21 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101344

Rudolf W van Olden, Christophe Lo Bianco, Keith W Dilly, Marina Savelieva, Siyan Xu, Aloys Tijsma, Carel van Baalen, Harsh Sharma, Nayla Mumneh

{"title":"Adeno-associated virus serotype 9 antibodies in neonates and young children: Seroprevalence and kinetics.","authors":"Rudolf W van Olden, Christophe Lo Bianco, Keith W Dilly, Marina Savelieva, Siyan Xu, Aloys Tijsma, Carel van Baalen, Harsh Sharma, Nayla Mumneh","doi":"10.1016/j.omtm.2024.101344","DOIUrl":"10.1016/j.omtm.2024.101344","url":null,"abstract":"Gene therapies such as onasemnogene abeparvovec for spinal muscular atrophy (SMA) utilize adeno-associated virus 9 (AAV9) for targeted gene delivery, which requires an AAV9 antibody (AAV9-Ab) immunoglobulin G (IgG) ≤1:50 titer threshold. This retrospective cohort study evaluated age-related AAV9-Ab IgG seroprevalence for patients with SMA (part 1) and AAV9-Ab IgG kinetics and time to 1:50 titer threshold in newborns with elevated AAV9-Ab IgG titers (≥1:100) (part 2). A semi-quantitative ELISA assay was used in part 1 (N = 1,323 patients). For patients aged <12 months, 3.9% (n/N = 31/795) had elevated AAV9-Ab IgG titers (≥1:100); prevalence declined with age. In part 2, a new quantitative ELISA (linear mixed effects model) described continuous AAV9-Ab IgG concentrations for patients with initial titers ≥1:100. AAV9-Ab IgG concentrations waned according to first-order kinetics (58 samples; N = 18 patients). The model-based estimation of the AAV9-Ab IgG average half-life was 41 (95% CI, 38-44) days. Based on visualization, 200 ELISA units/mL was a reasonable approximate for the 1:50 titer threshold. In conclusion, initially elevated titers ≥1:100 in newborn patients declined with age. The new quantitative ELISA may allow for quantification of time to threshold for AAV9-Ab IgG retesting for onasemnogene abeparvovec treatment, leading to treatment as early as possible for patients with SMA.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101344"},"PeriodicalIF":4.6,"publicationDate":"2024-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11866128/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143525250","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Non-encapsidated miRNA contaminants found in AAV preparations 在 AAV 制剂中发现的非包囊 miRNA 杂质

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-20 DOI: 10.1016/j.omtm.2024.101336

Mark A. Brimble, Stephen M. Winston

引用次数: 0

A novel AAV9-dual microRNA- vector targeting GRIK2 in the hippocampus as a treatment for mesial temporal lobe epilepsy 以海马中的 GRIK2 为靶点的新型 AAV9 双 microRNA 载体可用于治疗中位颞叶癫痫病

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-16 DOI: 10.1016/j.omtm.2024.101342

Stéphane J. Baudouin, April Giles, Nick Pearson, Severine Deforges, Chenxia He, Céline Boileau, Nicolas Partouche, Andreas Borta, Justine Gautron, Morgane Wartel, Irena Bočkaj, Didier Scavarda, Fabrice Bartolomei, Guillaume Penchet, Jérôme Aupy, Jennifer Sims, Jared Smith, Andrew Mercer, Olivier Danos, Christophe Mulle, Richard Porter

{"title":"A novel AAV9-dual microRNA- vector targeting GRIK2 in the hippocampus as a treatment for mesial temporal lobe epilepsy","authors":"Stéphane J. Baudouin, April Giles, Nick Pearson, Severine Deforges, Chenxia He, Céline Boileau, Nicolas Partouche, Andreas Borta, Justine Gautron, Morgane Wartel, Irena Bočkaj, Didier Scavarda, Fabrice Bartolomei, Guillaume Penchet, Jérôme Aupy, Jennifer Sims, Jared Smith, Andrew Mercer, Olivier Danos, Christophe Mulle, Richard Porter","doi":"10.1016/j.omtm.2024.101342","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101342","url":null,"abstract":"Mesial temporal lobe epilepsy (mTLE) is the most prevalent type of epilepsy in adults. First and subsequent generations of anti-epileptic therapy regimens fail to decrease seizures in a large number of patients suffering from mTLE, leaving surgical ablation of part of the hippocampus as the only therapeutic option to potentially reach seizure freedom. GluK2 has recently been identified as a promising target for the treatment of mTLE using gene therapy. Here, we engineered an adeno-associated virus serotype 9 vector expressing a cluster of two synthetic microRNAs (miRNAs), expressed from the human synapsin promoter, that target GRIK2 mRNA. Intra-hippocampal delivery of this vector in a mouse model of mTLE significantly reduced GRIK2 expression and daily seizure frequency. This treatment also improved the animals' health, reduced their anxiety, and restored working memory. Focal administration of the vector to the hippocampus of cynomolgus monkeys in GLP toxicology studies led to the selective transduction of hippocampal neurons with little exposure elsewhere in the brain and no transduction outside the central nervous system. Expression of miRNAs in hippocampal neurons resulted in substantially decreased GRIK2 mRNA expression. These data suggest that the intra-hippocampal delivery of a GMP-grade AAV9 coding for a synthetic miRNAs targeting GRIK2 is a promising treatment strategy for mTLE.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"15 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252865","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Optimal different adeno-associated virus capsid/promoter combinations to target specific cell types in the common marmoset cerebral cortex 针对普通狨猴大脑皮层特定细胞类型的最佳腺相关病毒壳/启动子组合

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-13 DOI: 10.1016/j.omtm.2024.101337

Yasunori Matsuzaki, Yuuki Fukai, Ayumu Konno, Hirokazu Hirai

{"title":"Optimal different adeno-associated virus capsid/promoter combinations to target specific cell types in the common marmoset cerebral cortex","authors":"Yasunori Matsuzaki, Yuuki Fukai, Ayumu Konno, Hirokazu Hirai","doi":"10.1016/j.omtm.2024.101337","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101337","url":null,"abstract":"To achieve cell type-specific gene expression, using target cell type-tropic different adeno-associated virus (AAV) capsids is advantageous. However, their tropism across brain cell types in nonhuman primates has not been fully elucidated. We assessed the tropism of nine AAV serotype capsids (AAV1, 2, 5, 6, 7, 8, 9, rh10, and DJ) expressing enhanced green fluorescent protein (EGFP) by chicken β-actin hybrid (CBh) promoter in marmoset cerebral cortical cells. All nine AAV capsid vectors, especially AAV9 and AAVrh10, caused highly neuron-selective EGFP expression. Some AAV capsids, including AAV5, induced EGFP expression to a lesser extent in oligodendrocytes. Different ubiquitous cytomegalovirus (CMV) and CMV early enhancer/chicken β actin (CAG) promoters exhibited similar neuron-predominant transgene expression. Conversely, all nine AAV capsid vectors with the astrocyte-specific hGFA(ABC1D) promoter selectively expressed EGFP in astrocytes, except AAV5, which modestly expressed EGFP in oligodendrocytes. Oligodendrocyte-specific mouse myelin basic protein (mMBP) promoter in AAV5 vectors expressed EGFP in oligodendrocytes specifically and efficiently. The following are optimal combinations of capsids and promoters for cell type-specific expression: AAV9 or AAVrh10 and ubiquitous CBh or CMV promoter for neuron-specific transgene expression; AAV2 or AAV7 and hGFA(ABC1D) promoters for astrocyte-specific transgene expression; and AAV5 and mMBP promoters for oligodendrocyte-specific transgene expression.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"27 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142197239","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0