Human gene therapy最新文献

{"title":"Quantification of Linear Polyethylenimines in Recombinant Adeno-Associated Virus by High-Performance Liquid Chromatography with Charged Aerosol Detection.","authors":"Amelia W Paine, Stacey Nash, Nicolsond Nelson, Eric Crain, Xiaohui Lu, James Warren, Jin Seon Park","doi":"10.1177/10430342251411041","DOIUrl":"10.1177/10430342251411041","url":null,"abstract":"<p><p>Polyethylenimine (PEI) is widely employed as a transfection reagent in recombinant adeno-associated virus (rAAV) manufacturing, but it must be removed from the final product due to its potential toxicity. Accurate quantification of PEI in complex biological matrices such as rAAVs is challenging, largely because the strong electrostatic attraction between PEI and nucleic acids can hinder the accuracy of its quantification. Here, we report a robust high-performance liquid chromatography method with charged aerosol detection for the quantification of residual linear PEI in purified AAV samples. Sample preparation includes treatment with trifluoroacetic acid and hydrochloric acid at 60°C to denature capsid protein, disrupt PEI-DNA polyplexes, and hydrolyze nucleic acids. The method achieves a limit of detection of 5 µg/mL and a limit of quantitation of 10 µg/mL in spike-and-recovery studies, with quantification confirmed via visual peak identification. This approach enables sensitive, specific, and reproducible PEI measurement and provides a valuable tool for process monitoring and quality control in gene therapy manufacturing.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"365-371"},"PeriodicalIF":4.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146131664","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Brain-Directed AAV Gene Therapy Rescues a Mouse Model of the CLN5 Form of Neuronal Ceroid Lipofuscinosis Disease and Normalizes a Blood Plasma Biomarker of Neurodegeneration.","authors":"Wenfei Liu, Amy F Geard, Giulia Massaro, Michael P Hughes, Mikel Aristorena, Oliver Coombe-Tennant, Liangyuan Xu, Olha Semenyuk, Reuben Bush, Danielle Te Vruchte, David Priestman, Rhiannon Laban, Elena Veleva, Amanda J Heslegrave, Henrik Zetterberg, Frances M Platt, Alexander J Smith, Sara E Mole, Robin R Ali, Ahad A Rahim","doi":"10.1177/10430342251403448","DOIUrl":"10.1177/10430342251403448","url":null,"abstract":"<p><p>CLN5 disease, caused by mutations in the <i>CLN5</i> gene, is a form of neuronal ceroid lipofuscinoses (Batten disease). Patients suffer progressive motor dysfunction, vision loss, seizures, and dementia, leading to premature death. Here, we report a preclinical study of AAV9-mediated gene therapy in a <i>Cln5</i>^-/- mouse model. Single-dose AAV9 carrying human <i>CLN5</i> driven by the CAG or human synapsin 1 promoter (hSYN) was administered via intracerebroventricular injection into neonatal and juvenile <i>Cln5</i>^-/- mice. Treatment efficacy was evaluated by assessment of neurodegeneration, neuroinflammation, locomotor function, and survival. AAV9 expressing <i>CLN5</i> driven by the hSYN promoter significantly alleviated neurodegeneration, improved biochemical and glycosphingolipid profiles, neuropathological and locomotor function, and extended lifespan of the <i>Cln5</i>^-/- mice. However, gene transfer employing the CAG promoter demonstrated limited therapeutic efficacy. Furthermore, delayed intervention in juveniles provided superior therapeutic response compared with early neonatal intervention and normalized lifespan. Finally, blood plasma neurofilament light that is significantly elevated in the <i>Cln5</i>^-/- mice is restored to normal wildtype levels following treatment. These results indicate that brain-directed adeno-associated virus (AAV) gene therapy could be a promising treatment strategy for CLN5 disease and efficacy might be monitored using a noninvasive blood plasma biomarker.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"241-261"},"PeriodicalIF":4.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145850108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Maximizing Access to Cell and Gene Therapy for Patients with Rare Diseases.","authors":"Terence R Flotte, Guangping Gao","doi":"10.1177/10430342261422716","DOIUrl":"10.1177/10430342261422716","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"163-164"},"PeriodicalIF":4.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146118890","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ikarovec Signs Up-to-$1B+ Exclusive Worldwide Option for VectorBuilder AAV Capsid Technology.","authors":"Alex Philippidis","doi":"10.1177/10430342261423148","DOIUrl":"https://doi.org/10.1177/10430342261423148","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":"37 5-6","pages":"165-169"},"PeriodicalIF":4.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147432472","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Learnings from Patient Mortality after Delandistrogene Moxeparvovec Administration: A Report of Two Cases and Expert Committee Considerations for Future Mitigation and Management.","authors":"Perry B Shieh, Crystal Proud, Tamir Diamond, Catherine A Chapin, Jawad Ahmad, Alan D Salama, Carsten G Bönnemann, Jonathan Soslow, Barry J Byrne, Aravindhan Veerapandiyan, John F Brandsema, Susan Matesanz, Benjamin J Samelson-Jones, Benjamin J Wilkins, Marianne Gerber, Kara E Godwin Wild, Stefanie Mason, Damon Asher, Craig M McDonald, Jerry R Mendell","doi":"10.1177/10430342261423168","DOIUrl":"10.1177/10430342261423168","url":null,"abstract":"<p><p>Acute liver injury (ALI) is a recognized adverse event with adeno-associated virus (AAV)-based gene therapies, including delandistrogene moxeparvovec, an AAVrh74-based gene therapy for Duchenne muscular dystrophy. Progression of ALI to acute liver failure (ALF) is rare. In clinical trials, ALF was defined as an international normalized ratio ≥1.5 with encephalopathy and/or ascites, occurring <26 weeks since product/treatment exposure, and in the absence of identified preexisting liver disease as assessed by the treating investigator. The two cases presented here represent the only known instances of ALF following delandistrogene moxeparvovec to date, both resulting in fatal outcomes. Both cases occurred in nonambulatory patients (ages 15 and 16 years). Both patients exhibited abrupt elevations in aminotransferases from baseline approximately 4 weeks after delandistrogene moxeparvovec administration. Over the subsequent weeks and despite interventions, aminotransferases, including gamma-glutamyl transferase (GGT), declined concurrently with a continued rise in total bilirubin, consistent with severe hepatocellular injury and a harbinger of ALF. An interdisciplinary expert panel was convened and concluded that no single biomarker or clinical feature consistently predicted the rapid progression of severe ALI to ALF. Advisors recommended basing treatment intervention for severe ALI on the rate and magnitude of changes from baseline in liver biomarkers (<i>e.g.,</i> 2-3× increase in alanine aminotransferase within ≤1 week). For these severe cases, most advisors recommended intravenous methylprednisolone as the initial treatment whereas typical ALI may be managed with oral corticosteroid adjustment or may resolve with time, as seen in the clinical trial experience. Advisors hypothesized that delandistrogene moxeparvovec-related ALI is T-cell-mediated and discussed the potential of additional immunosuppression. Advisors also recommended enhancing baseline evaluation for hepatic comorbidities with lipid profiling, transient elastography, and abdominal ultrasound to further study potential risk factors for ALI progressing to ALF. Finally, there was strong support for generating additional real-world evidence and conducting prospective clinical trials to inform clinical management of ALI in practice.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"262-279"},"PeriodicalIF":4.6,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13123743/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146179456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multivariate Process Optimization for Fixed-Bed Bioreactor-Based AAV Production Improves Total Batch Yield.","authors":"Ruchita Selot, Ashish Khaparde, Sharath Babu G R, Chitra Gopinath, Trailokyanath Panigrahi, Subhradeep Sarkar, Joy Elvin Dhinakar, Riya Patra, Priyalakshmi Panikker, Arkasubhra Ghosh","doi":"10.1177/10430342251396547","DOIUrl":"10.1177/10430342251396547","url":null,"abstract":"<p><p>Adeno-associated viral vectors (AAVs) are promising tools for gene therapy. However, scaling up the production of AAVs to produce high-quality vectors at high yields for clinical purposes has proven to be challenging. In the present study, we optimized the production process of AAV in a fixed-bed bioreactor using transient transfection in adherent HEK-293T cells. We systematically optimized the key process parameters, namely cell seeding density, cell density at transfection, and DNA-to-cell ratio, based on the yield obtained, starting from a prototype batch, followed by ten batch runs. Here, we packaged a reporter gene (enhanced green fluorescent protein) and a therapeutic gene (lysyl oxidase) into AAV9 capsids as part of our process development program to be applied for future current Good Manufacturing Practices production and clinical trial application. Throughout the experiments, media conditions, transfection processes, and mechanical parameters were kept identical, while monitoring pH, dissolved oxygen, and media glucose concentration during a production process of approximately 10 days. We demonstrate that by optimizing these parameters, the fixed-bed bioreactor was able to support as many as 1.6-2.8 × 10⁶ cells/carrier strip, up to 3 × 10⁹ cells/m² bioreactor. Through this multivariate optimization process, we increased viral yield by about 7.6-fold (range of 5.7-10.4-fold for the optimized process runs) over the prototype batch. The total AAV vector yield average was 2.3 × 10¹⁴ vg (range 1.1 × 10¹⁴ vg to 4.95 × 10¹⁴ vg), corresponding to an average per cell yield of 1.4 × 10⁵ vg/cell (range 0.85 × 10⁵-2.46 × 10⁵vg/cell). In conclusion, our findings highlight that optimizing process parameters in a fixed-bed bioreactor presents a promising strategy for scalable and cost-effective AAV vector production.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"280-289"},"PeriodicalIF":4.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145587289","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Systemic Administration of AAV9-G266A with Focused Ultrasound Enables Robust Brain Transduction with Minimal Liver Transduction.","authors":"Andrew T Nelson, Aidan Smith, Colleen T Curley, Brian Kelly, Ahil N Ganesh, Drew Peterson, Emily Fabyanic, Eric Kostuk, Thomas Weber, Esteban A Engel","doi":"10.1177/10430342251411044","DOIUrl":"10.1177/10430342251411044","url":null,"abstract":"<p><p>Systemic delivery of adeno-associated viral (AAV) vectors is a promising approach for brain gene therapy, particularly in combination with emerging techniques such as focused ultrasound (FUS), which can transiently and noninvasively open the blood-brain barrier to facilitate delivery of AAVs to the brain. However, off-target vector accumulation, particularly in the liver, remains a significant safety concern. To address this, we introduced a mutation into variable region 1 of the AAV9 capsid (G266A) and assessed its effect on liver and brain transduction, testing both direct intraparenchymal injection and intravenous injection with FUS. Interestingly, we found that G266A mutation strongly decreased liver transduction while having only a modest effect on transduction of the brain and other tissues. We also found that the G266A mutation had minimal impact on cell attachment and uptake but strongly decreased binding of the capsid to both human and mouse AAV receptor (AAVR), which likely explains the decrease in liver transduction. Overall, our findings suggest that the G266A mutation and modification of AAVR binding could be a useful strategy to mitigate liver toxicity associated with systemic brain gene therapy.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"199-209"},"PeriodicalIF":4.6,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146142189","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CRISPR in Medicine: A Systematic Review of Clinical Trials and Therapeutic Applications.","authors":"Mohammad Rahmanian, Mohadeseh Khoshandam, Marziyeh Mousazadeh, Piao Yang, Hossein Soltaninejad, Pouya Karami Dehkordi, Majid Sadeghizadeh, Mohammad Taghi Hedayati Goudarzi, Amir Hossein Azimi, Mohsen Sheykhhasan","doi":"10.1177/10430342251409714","DOIUrl":"10.1177/10430342251409714","url":null,"abstract":"<p><p>Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR associated protein 9 (CRISPR/Cas9) technology has become a revolutionary tool in medicine, offering substantial potential for treating a wide range of diseases, including hematological disorders, cancers, genetic conditions, and ophthalmological diseases. This systematic review evaluates the efficacy, safety, and applicability of CRISPR/Cas9 in clinical trials. A comprehensive search of the PubMed, Scopus, Web of Science, and Cochrane databases was conducted. All studies, up to November 2024, meeting the eligibility criteria assessing the application of CRISPR for the treatment of diseases were included. A quality assessment of the included studies was conducted using the Cochrane risk of bias tool. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 statement for systematic reviews and meta-analyses was followed, and a total of 17 studies were included. This systematic review of CRISPR/Cas9 technology focused on its effectiveness and safety across various diseases. In nonmalignant hematological disorders, CRISPR successfully treated β-thalassemia and sickle cell disease, resulting in high transfusion independence and the elimination of disease crises. In malignant hematological disorders, B-cell acute lymphoblastic leukemia, CRISPR-engineered chimeric antigen receptor T (CAR-T) cells achieved an 83.3% complete remission rate. Furthermore, CRISPR-based CAR-T cells showed promising results in B-cell non-Hodgkin's lymphoma. In oncology, lung cancer and other solid tumors are among the diseases that have been safely engineered using CRISPR gene editing technology. For genetic disorders, CRISPR improved vision in retinal degeneration and reduced symptoms in hereditary angioedema and transthyretin amyloidosis with mild side effects. The results demonstrated CRISPR's potential across a wide range of conditions. In conclusion, the findings underscore the potential role of CRISPR/Cas9 technology across a wide range of diseases. However, challenges remain, including optimizing delivery systems, minimizing off-target effects, addressing immunogenicity concerns, and ethical considerations.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":"37 5-6","pages":"170-182"},"PeriodicalIF":4.6,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147432447","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Prenatal Intraportal Delivery of Polymeric Nanoparticles to Fetal Rhesus Monkeys (<i>Macaca mulatta</i>).","authors":"Alexandra S Piotrowski-Daspit, Anna Y Lynn, David A Eaton, Laura G Bracaglia, Arianna I Markey, Ryland D Mortlock, David H Stitelman, Michele L Martinez, Charles C Lee, Harvey J Kliman, Peter M Glazer, Alice F Tarantal, W Mark Saltzman","doi":"10.1177/10430342251414881","DOIUrl":"10.1177/10430342251414881","url":null,"abstract":"<p><p>Preliminary investigations focused on biodistribution of polymeric nanoparticles (NPs) shortly after ultrasound-guided delivery via the portal vein in early second-trimester fetal rhesus monkeys. Results demonstrated that poly(lactic-co-glycolic acid) (PLGA) NPs (<i>N</i> = 3; 3 mg administered at 75-80 days gestational age) and poly(amine-co-ester)-polyethylene glycol (PACE-PEG) NPs (<i>N</i> = 3; 3 mg at 75-80 days gestational age) distributed to fetal tissues when assessed 24 h post-administration. No adverse findings were observed. PLGA NPs were found primarily in the fetal liver and spleen, whereas PACE-PEG NPs showed more widespread biodistribution to a range of anatomical sites. In another fetal subset with PACE-PEG NPs (<i>N</i> = 2; 90 days gestational age) assessed within 48-h post-administration, results demonstrated enhanced green fluorescent protein reporter mRNA expression in select tissues. These early-stage short-term studies suggest that polymeric NPs, particularly those composed of PACE-PEG, can be safely administered and are potential candidates for fetal delivery of therapeutic nucleic acids. While preliminary, these studies provide evidence to support further investigations in this species to address long-term safety and efficiency.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"189-198"},"PeriodicalIF":4.6,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12978997/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146124779","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Efficient LAMA1 Gene Activation by Epigenome Editing as a Therapeutic Approach for LAMA2-CMD.","authors":"Yuanbo Qin, Talha Akbulut, Rajakumar Mandraju, Keith Connolly, John Bechill, Farzaneh Assadian, Claudia Foster, Alison Shottek, Seth Levy, Jamie Benoit, Tetsuya Yamagata","doi":"10.1177/10430342251401873","DOIUrl":"10.1177/10430342251401873","url":null,"abstract":"<p><p>Epigenome editing technology holds great promise for treating diverse genetic disorders. In this study, we demonstrate epigenetic activation of the <i>LAMA1</i> gene for the treatment of <i>LAMA2</i>-CMD, a severe congenital muscle dystrophy (CMD) caused by biallelic mutations in the <i>LAMA2</i> gene. <i>LAMA1</i> is a sister homolog that is known to compensate for the function of <i>LAMA2</i>. However, supplementing <i>LAMA1</i> or <i>LAMA2</i> gene via viral platform is not feasible due to the large size of their coding sequences. Through a single administration of our adeno-associated virus (AAV) vector encoding all the necessary elements for epigenetic activation, we observed significant <i>LAMA1</i> gene upregulation and phenotype improvements in mouse disease models. The muscle-tropic AAV capsid exhibited desired vector biodistribution and promising pharmacodynamics with good safety profiles in 2-year-old juvenile nonhuman primates (NHPs). Moreover, administration to 8-month-old infant NHPs demonstrated superior pharmacodynamics compared with 2-year-old juveniles, even at half the dose. Our approach holds broad applicability for a range of loss-of-function genetic disorders and could offer a therapeutic breakthrough where active epigenome offers clinical benefit.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"210-222"},"PeriodicalIF":4.0,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145819021","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}