Gene Therapy最新文献

Genetic mutations in HSV-1 replication-defective vectors: Implications for their safety in gene therapy applications. HSV-1复制缺陷载体的基因突变：对其在基因治疗应用中的安全性的影响。

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-09-28 DOI: 10.1038/s41434-025-00566-1

Stefano Cattaneo, Barbara Bettegazzi, Selene Ingusci, Gianluca Verlengia, Tascini Anna Sofia, Zucchini Silvia, Franca Codazzi, Marco J Morelli, Marco Marzulli, Joseph C Glorioso, Michele Simonato

{"title":"Genetic mutations in HSV-1 replication-defective vectors: Implications for their safety in gene therapy applications.","authors":"Stefano Cattaneo, Barbara Bettegazzi, Selene Ingusci, Gianluca Verlengia, Tascini Anna Sofia, Zucchini Silvia, Franca Codazzi, Marco J Morelli, Marco Marzulli, Joseph C Glorioso, Michele Simonato","doi":"10.1038/s41434-025-00566-1","DOIUrl":"https://doi.org/10.1038/s41434-025-00566-1","url":null,"abstract":"Beyond its well-known role in orofacial recurrent infections, HSV-1 has garnered significant attention in neuroscience for contrasting reasons. On one hand, it has been found to be involved in neurodegenerative processes; on the other, it may represent a versatile platform for gene therapy of brain diseases, due to its large genome that enables the delivery of sizable or multiple genes. These opposite features underscore the importance of understanding HSV-1 interactions with neural tissues in view of its employment as a gene therapy platform. We recently developed a new generation of highly defective backbones that proved very efficient and safe after direct injection in the brain parenchyma. Here we aimed at probing in depth the safety of viral batches that lack obvious unwanted (specifically, fusogenic) activities during production and, therefore, may escape negative selection. We employed whole-genome sequencing, electrophysiology, and viral engineering to compare different viral batches. We identified mutations (in particular A to I at position 549 in the UL27 gene) that confer fusogenic capacity to the envelop glycoprotein gB, inducing a hyperexcitable phenotype in transduced neurons. Such syncytial variants should be identified and avoided for any application of HSV-1 vectors implicating their direct injection in the nervous system.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2025-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145185499","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}

引用次数: 0

Overcoming matrix effects in AAV neutralization assays with a constant serum concentration approach. 用恒血清浓度法克服AAV中和试验中的基质效应。

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-09-26 DOI: 10.1038/s41434-025-00567-0

Beatrix Kovács, Viktória Szabó, Domonkos Horváth, Attila Balázs Dobos, Zoltán Zsolt Nagy, Wim Vanduffel, Zsuzsanna Szemlaky, Áron Szepesi, István Ulbert, Balázs Rózsa, Dániel Hillier

{"title":"Overcoming matrix effects in AAV neutralization assays with a constant serum concentration approach.","authors":"Beatrix Kovács, Viktória Szabó, Domonkos Horváth, Attila Balázs Dobos, Zoltán Zsolt Nagy, Wim Vanduffel, Zsuzsanna Szemlaky, Áron Szepesi, István Ulbert, Balázs Rózsa, Dániel Hillier","doi":"10.1038/s41434-025-00567-0","DOIUrl":"https://doi.org/10.1038/s41434-025-00567-0","url":null,"abstract":"Sensitive quantification of adeno-associated virus (AAV) neutralizing antibodies (NAbs) is essential for gene therapy success. Conventional cell-based transduction inhibition assays often encounter matrix-induced artifacts resulting from variable serum content across dilutions, which artificially inflate transduction baselines and mask partial neutralization. To address this challenge, we developed the constant serum concentration (CSC) assay, which maintains constant serum levels across dilutions to stabilize assay baselines and enhance NAb detection sensitivity. Using human sera across multiple AAV serotypes, we demonstrated that CSC reclassified up to 21.7% of samples classified as non-neutralizing by a conventional variable serum concentration (VSC) assay format. This improved sensitivity was validated using monoclonal antibody and multi-species serum test benchmarks and enhanced the reliability of seronegative control pool selection. Importantly, CSC detected persistent seropositivity in preclinical seroreversion models up to one year longer than a conventional VSC assay. Since even low-level neutralizing antibodies can significantly impact gene therapy efficacy, these findings have direct implications for optimizing AAV redosing strategies and refining patient stratification. By addressing fundamental limitations in NAb quantification while maintaining procedural simplicity, the CSC assay provides crucial insights into antibody persistence with translational relevance across species and clinical settings.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2025-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145174670","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}

引用次数: 0

Correction: PPARγ is essential for protection against nonalcoholic steatohepatitis. 更正：PPARγ对预防非酒精性脂肪性肝炎至关重要。

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-09-26 DOI: 10.1038/s41434-025-00568-z

C W Wu, E S H Chu, C N Y Lam, A S L Cheng, C W Lee, V W S Wong, J J Y Sung, J Yu

引用次数: 0

Correction: Identification of AAV variants with improved transduction of human vascular endothelial cells by screening AAV capsid libraries in non-human primates. 更正：通过筛选非人灵长类动物的AAV衣壳文库，鉴定具有改进的人血管内皮细胞转导的AAV变体。

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-09-05 DOI: 10.1038/s41434-025-00565-2

Maria Stamataki, Julia Lüschow, Christina Schlumbohm, Malik Alawi, Lars Lunding, Eberhard Fuchs, Martin Trepel, Markus Schwaninger, Jakob Körbelin

引用次数: 0

AAV-mediated MUC5AC siRNA delivery to prevent mucociliary dysfunction in asthma. aav介导的MUC5AC siRNA递送预防哮喘患者粘膜纤毛功能障碍。

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-08-23 DOI: 10.1038/s41434-025-00564-3

Sahana Kumar, Maria Corkran, Yahya Cheema, Margaret A Scull, Gregg A Duncan

{"title":"AAV-mediated MUC5AC siRNA delivery to prevent mucociliary dysfunction in asthma.","authors":"Sahana Kumar, Maria Corkran, Yahya Cheema, Margaret A Scull, Gregg A Duncan","doi":"10.1038/s41434-025-00564-3","DOIUrl":"https://doi.org/10.1038/s41434-025-00564-3","url":null,"abstract":"The main structural components of mucus produced in the lung are mucin 5B (MUC5B) and mucin 5AC (MUC5AC) where a relatively higher expression of MUC5B is typical in health. In the lungs of individuals with asthma, there is a shift from MUC5B to MUC5AC as the predominantly secreted mucin which has been shown to impair mucociliary clearance (MCC) and increase airway mucus plug formation. Given its role in asthmatic lung disease, MUC5AC represents a potential therapeutic target where a gene delivery approach could be leveraged to modulate its expression. For these purposes, we explored adeno-associated virus serotype 6 (AAV6), as a viral gene vector to transduce airway epithelial cells and reduce MUC5AC expression via siRNA delivery. We confirmed that AAV6 was able to transduce epithelial cells in vitro as well as in the airways of healthy mice in vivo with high transgene expression in mucus-secreting goblet cells. Using multiple particle tracking analysis, we observed that AAV6 was capable of penetrating both normal and MUC5AC-enriched mucus barriers. AAV6 carrying MUC5AC-targeting siRNA was evaluated as a prophylactic treatment in HAE cell cultures before IL-13 challenge. IL-13 stimulated HAE cultures treated with AAV6-MUC5AC siRNA had significantly reduced MUC5AC mRNA and protein expression compared to untreated controls. Mucociliary transport in IL-13 stimulated HAE cultures was also maintained and comparable to healthy controls following AAV6-MUC5AC siRNA treatment. Together, these findings support that AAV6 may be used as an inhaled gene therapy to suppress MUC5AC overexpression and restore normal airway clearance function in asthma.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2025-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144951532","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}

引用次数: 0

Identification of AAV variants with improved transduction of human vascular endothelial cells by screening AAV capsid libraries in non-human primates. 通过筛选非人灵长类动物AAV衣壳文库，鉴定具有改善人血管内皮细胞转导的AAV变体。

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-08-22 DOI: 10.1038/s41434-025-00563-4

Maria Stamataki, Julia Lüschow, Christina Schlumbohm, Malik Alawi, Lars Lunding, Eberhard Fuchs, Martin Trepel, Markus Schwaninger, Jakob Körbelin

{"title":"Identification of AAV variants with improved transduction of human vascular endothelial cells by screening AAV capsid libraries in non-human primates.","authors":"Maria Stamataki, Julia Lüschow, Christina Schlumbohm, Malik Alawi, Lars Lunding, Eberhard Fuchs, Martin Trepel, Markus Schwaninger, Jakob Körbelin","doi":"10.1038/s41434-025-00563-4","DOIUrl":"10.1038/s41434-025-00563-4","url":null,"abstract":"The development of targeted vector systems for gene therapy has made impressive progress during the last decade. Promising vector candidates were identified by screening large pools of adeno-associated virus (AAV) mutants in small animal models. However, it became apparent that targeted AAV mutants isolated from rodents may not function in humans as the tropism of individual AAV mutants can differ between species. To identify novel vascular-targeted AAV capsid mutants suitable for treating human patients, we generated a set of AAV2 display peptide libraries and screened them in the common marmoset, a non-human primate. To evaluate the impact of different AAV library production methods, progress of the screening process was monitored by next generation sequencing. Particle distribution and enrichment was compared between different AAV libraries and selection rounds. We observed enrichment of AAV variants in the brain and other well-perfused organs (lung, heart, kidney) potentially mediated by high capsid affinity for the vascular endothelium in general. In vitro experiments on primary human microvascular endothelial cells isolated from a set of different organs (brain, heart, lung, liver, kidney and spleen) confirmed superior transduction of a selected AAV variant displaying the \"DWP\" amino acid sequence motif compared to natural AAV serotypes 1-9.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2025-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144951564","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}

引用次数: 0

Gene therapy restores auditory function and rescues damaged inner hair cells in an aged Vglut3 knockout mouse model. 在Vglut3基因敲除小鼠模型中，基因治疗可恢复听觉功能并挽救受损的内毛细胞。

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-08-21 DOI: 10.1038/s41434-025-00558-1

Xingle Zhao, Hongen Xu, Chengyu Lian, Shousen Hu, Yue Zhao, Jia Wang, Rongqun Zhai, Mihuan Yang, Yuanjing Zhang, Wei Lu, Wenxue Tang, Liang Wang

{"title":"Gene therapy restores auditory function and rescues damaged inner hair cells in an aged Vglut3 knockout mouse model.","authors":"Xingle Zhao, Hongen Xu, Chengyu Lian, Shousen Hu, Yue Zhao, Jia Wang, Rongqun Zhai, Mihuan Yang, Yuanjing Zhang, Wei Lu, Wenxue Tang, Liang Wang","doi":"10.1038/s41434-025-00558-1","DOIUrl":"https://doi.org/10.1038/s41434-025-00558-1","url":null,"abstract":"Vesicular glutamate transporter 3 (VGLUT3) is prominently expressed in the inner hair cells of the cochlea, playing a vital role in auditory signal transmission to the brain. Previous studies have shown that Vglut3 gene knockout in mice causes severe sensorineural hearing loss without affecting hair cell integrity. However, the cochlear structure of the aged Vglut3KO remains inadequately explored. In this study, we analyzed the cochlear structure of aged Vglut3KO mice, revealing significant degeneration of inner hair cells, synapses, and stereocilia. To explore the potential of gene therapy to restore cochlear structure, we employed AAV8 vectors to express Vglut3 in the cochleae of 5-week-old Vglut3KO mice. Twenty-seven weeks post-injection, we conducted a series of experiments to evaluate the efficacy of our gene therapy approach. Auditory brainstem response (ABR) testing demonstrated restoration of auditory function following gene therapy. Immunohistochemical staining and scanning electron microscopy (SEM) analysis revealed substantial recovery of inner hair cells and stereocilia post-injection. Our findings provide important insights into the development of novel therapeutic strategies for age-related hearing loss.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2025-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144951497","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}

引用次数: 0

Rapid detection of AAV8 binding antibodies in gene therapy candidates: development of a point-of-care approach. 基因治疗候选药物中AAV8结合抗体的快速检测：一种即时护理方法的发展。

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-08-21 DOI: 10.1038/s41434-025-00559-0

Alexander Kozikowski, Qing Wang, Cheng Yang, Neil Gordon, Kristina M Ciociola, Asanka Yapa, Claudia Villa, Paul Lambotte, Thomas Pisani, Javan Esfandiari, Angelo H Gunasekera

{"title":"Rapid detection of AAV8 binding antibodies in gene therapy candidates: development of a point-of-care approach.","authors":"Alexander Kozikowski, Qing Wang, Cheng Yang, Neil Gordon, Kristina M Ciociola, Asanka Yapa, Claudia Villa, Paul Lambotte, Thomas Pisani, Javan Esfandiari, Angelo H Gunasekera","doi":"10.1038/s41434-025-00559-0","DOIUrl":"https://doi.org/10.1038/s41434-025-00559-0","url":null,"abstract":"Preexisting anti-AAV antibodies pose a significant challenge to the success of Adeno-associated Virus (AAV) mediated gene therapies, as they can diminish therapeutic effectiveness, restrict patient eligibility for treatment, and cause serious health issues during treatment. This study introduces the first point-of-care (POC) test for the rapid, quantitative detection of AAV8 binding antibodies in patients' plasma, serum, and blood, leveraging Chembio's Dual Path Platform (DPP) technology. The DPP AAV8 Total Antibody (TAb) assay delivers results within 20 min from sample addition, with a dynamic range of 0-32 µg/ml when evaluated with purified human polyclonal antibodies that bind to AAV8, with reasonable specificity and sensitivity relative to Chembio's AAV8 TAb ELISA (R² = 0.90). Moreover, the assay demonstrated strong correlations with Chembio's AAV8 neutralizing antibody (NAb) ELISA and cell-based NAb assays (R² = 0.97 in plasma) (Cell-based assay adapted from BioAgilytix EU protocol). This rapid and reliable test can facilitate the screening of potential gene therapy patients for pre-existing antibodies that bind to AAV8 and assess their suitability for AAV8-mediated gene therapy.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2025-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144951525","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}

引用次数: 0

AAV microdystrophin gene replacement therapy for Duchenne muscular dystrophy: progress and prospects. AAV微营养不良蛋白基因替代治疗杜氏肌营养不良：进展与展望。

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-08-15 DOI: 10.1038/s41434-025-00561-6

Katarzyna Chwalenia, Vivi-Yun Feng, Nicole Hemmer, Hans J Friedrichsen, Ioulia Vorobieva, Matthew J A Wood, Thomas C Roberts

{"title":"AAV microdystrophin gene replacement therapy for Duchenne muscular dystrophy: progress and prospects.","authors":"Katarzyna Chwalenia, Vivi-Yun Feng, Nicole Hemmer, Hans J Friedrichsen, Ioulia Vorobieva, Matthew J A Wood, Thomas C Roberts","doi":"10.1038/s41434-025-00561-6","DOIUrl":"https://doi.org/10.1038/s41434-025-00561-6","url":null,"abstract":"Duchenne muscular dystrophy (DMD) is caused by pathogenic sequence variants occurring in the DMD gene which lead to the loss of the dystrophin protein, a molecular 'shock absorber' that protects muscle from contraction-induced injury. The large size of the dystrophin open reading frame precludes delivery of the full-length protein using a single adeno-associated virus (AAV) vector, which led to the development of internally-deleted dystrophin minigenes encoding partially-functional dystrophin. Indeed, five such microdystrophin therapies have been assessed in various clinical programmes. In 2023, Elevidys (Sarepta Therapeutics) received accelerated approval based on levels of dystrophin as a surrogate biomarker. In 2024, it received full approval despite unclear efficacy (i.e. not meeting primary or secondary outcomes in a phase 3 trial). Additionally, in 2025, two DMD individuals treated with Elevidys died after acute liver failure. A separate microdystrophin therapy, PF-06939926 (Pfizer) was discontinued for both efficacy and safety reasons (including the deaths of two clinical trial participants). Solid Biosciences, Genethon, REGENXBIO, and Insmed continue to develop microdystrophin therapies differing in transgene structure, promoter sequences, and AAV serotype. Here we describe recent progress in AAV-microdystrophin therapeutics development, and discuss the challenges facing such approaches, including pre-existing anti-capsid immunity, anti-transgene immunity, the unknown functionality of microdystrophin transgenes, transduction of muscle stem cells, and long-term transgene persistence.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144859100","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}

引用次数: 0

Correction: Expression of anti-amyloid CARs in microglia promotes efficient and selective phagocytosis of Aβ1‒42. 更正：小胶质细胞中抗淀粉样蛋白CARs的表达促进了Aβ1-42的有效和选择性吞噬。

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-08-15 DOI: 10.1038/s41434-025-00562-5

Christina N Heiss, Rebecca Riise, Eric Hanse, Stefanie Fruhwürth, Henrik Zetterberg, Andreas Björefeldt

引用次数: 0