Molecular Therapy-Methods & Clinical Development最新文献_第4页

Structural characterization and epitope mapping of the AAVX affinity purification ligand. AAVX 亲和纯化配体的结构特征和表位图谱。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-12 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101377

Mario Mietzsch, Manasi Kamat, Kari Basso, Paul Chipman, Juha T Huiskonen, Robert McKenna

{"title":"Structural characterization and epitope mapping of the AAVX affinity purification ligand.","authors":"Mario Mietzsch, Manasi Kamat, Kari Basso, Paul Chipman, Juha T Huiskonen, Robert McKenna","doi":"10.1016/j.omtm.2024.101377","DOIUrl":"10.1016/j.omtm.2024.101377","url":null,"abstract":"The application of adeno-associated virus (AAV) vectors in human gene therapies requires reproducible and homogeneous preparations for clinical efficacy and safety. For the AAV production process, often scalable affinity chromatography columns are utilized, such as the POROS CaptureSelect AAVX affinity resin, during downstream processing to ensure highly purified AAV vectors. The AAVX ligand is based on a camelid single-domain antibody capturing a wide range of recombinant AAV capsids. Described here is the identification of the AAV8 capsid epitope to AAVX at 2.3 Å resolution using cryo-electron microscopy. The ligand binds near the 5-fold axis of the capsid in a similar manner to the previously characterized AVB affinity ligand but does not conform to the capsid's icosahedral symmetry. The cross-reactivity of AAVX to other AAV capsids is achieved by primarily interacting with the peptide backbone of the AAV capsid's structurally conserved DE and HI loops. These observations will guide AAV capsid engineering efforts to retain the ability of future recombinant capsid designs to be purified using antibody-based affinity ligands.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101377"},"PeriodicalIF":4.6,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11638594/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142830827","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

Exogenous expression of ATP8, a mitochondrial encoded protein, from the nucleus in vivo. 线粒体编码蛋白ATP8在体内的外源表达。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-06 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101372

David V Begelman, Bhavna Dixit, Carly Truong, Christina D King, Mark A Watson, Birgit Schilling, Martin D Brand, Amutha Boominathan

{"title":"Exogenous expression of ATP8, a mitochondrial encoded protein, from the nucleus in vivo.","authors":"David V Begelman, Bhavna Dixit, Carly Truong, Christina D King, Mark A Watson, Birgit Schilling, Martin D Brand, Amutha Boominathan","doi":"10.1016/j.omtm.2024.101372","DOIUrl":"10.1016/j.omtm.2024.101372","url":null,"abstract":"Replicative errors, inefficient repair, and proximity to sites of reactive oxygen species production make mitochondrial DNA (mtDNA) susceptible to damage with time. We explore in vivo allotopic expression (re-engineering mitochondrial genes and expressing them from the nucleus) as an approach to rescue defects arising from mtDNA mutations. We used a mouse strain C57BL/6J(mtFVB) with a natural polymorphism (m.7778 G>T) in the mitochondrial ATP8 gene that encodes a protein subunit of the ATP synthase. We generated a transgenic mouse with an epitope-tagged recoded mitochondrial-targeted ATP8 gene expressed from the ROSA26 locus in the nucleus and used the C57BL/6J(mtFVB) strain to verify successful incorporation. The allotopically expressed ATP8 protein in transgenic mice was constitutively expressed across all tested tissues, successfully transported into the mitochondria, and incorporated into ATP synthase. The ATP synthase with transgene had similar activity to non-transgenic control, suggesting successful integration and function. Exogenous ATP8 protein had no negative impact on measured mitochondrial function, metabolism, or behavior. Successful allotopic expression of a mitochondrially encoded protein in vivo in a mammal is a step toward utilizing allotopic expression as a gene therapy in humans to repair physiological consequences of mtDNA defects that may accumulate in congenital mitochondrial diseases or with age.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101372"},"PeriodicalIF":4.6,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11629202/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142807731","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

Comparative assessment of the transduction efficiency and safety associated with the delivery of AAV9-GFP vector via lumbar puncture to cynomolgus macaques with and without anti-AAV9 pre-existing antibodies. AAV9-GFP载体经腰椎穿刺传递给具有和不具有抗aav9抗体的食蟹猴的转导效率和安全性的比较评估。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-06 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101371

Ghiabe H Guibinga, Janet Do, Binh Chu, Yin Gu, Rie Kikkawa, Xiaoguang Li, Fatih Ozsolak, Timothy MacLachlan

{"title":"Comparative assessment of the transduction efficiency and safety associated with the delivery of AAV9-GFP vector via lumbar puncture to cynomolgus macaques with and without anti-AAV9 pre-existing antibodies.","authors":"Ghiabe H Guibinga, Janet Do, Binh Chu, Yin Gu, Rie Kikkawa, Xiaoguang Li, Fatih Ozsolak, Timothy MacLachlan","doi":"10.1016/j.omtm.2024.101371","DOIUrl":"10.1016/j.omtm.2024.101371","url":null,"abstract":"Administration of AAV-based gene therapies into the intra-cerebrospinal fluid (CSF) compartments via routes such as lumbar puncture (LP) has been implemented as an alternative to intravenous dosing to target the CNS regions. This route enables lower doses, decreases systemic toxicity, and circumvents intravascular pre-existing anti-AAV antibodies. In this study, AAV9-GFP vectors were administered via LP to juvenile cynomolgus macaques with and without pre-existing serum anti-AAV9 antibodies at a 5.0 × 1013 vector genomes per mL (vg/mL) dose and examined for 28 days. CNS and peripheral tissues were surveyed for vector genome, mRNA, and protein expression. Histopathology, clinical pathology, and humoral immune response to the viral capsid and transgene were also assessed. In addition, serum and CSF samples were analyzed to examine 276 proteomic markers curated to evaluate neural injury, organ damage, and inflammatory response. This study reveals no noticeable difference in AAV9-mediated gene transfer in the CNS tissues in the two groups; however, differences were observed for endpoints such as liver enzyme activities, histopathology, and levels of protein markers in the serum and CSF. These findings provide a view into vector transduction efficiency and safety following LP-delivered AAV9 to juvenile cynomolgus macaques with and without pre-existing anti-AAV9 antibodies.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 4","pages":"101371"},"PeriodicalIF":4.6,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11664412/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142883091","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

Generation and maintenance of kidney and kidney cancer organoids from patient-derived material for drug development and precision oncology. 从患者来源的材料中生成和维持肾脏和肾癌类器官，用于药物开发和精确肿瘤学。

IF 4.6 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-11-05 eCollection Date: 2024-12-12 DOI: 10.1016/j.omtm.2024.101368

Jakub Gubala, Valentin Mieville, Daniel Benamran, Jean-Christophe Tille, Massimo Valerio, Patrycja Nowak-Sliwinska

引用次数: 0

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