Gene Therapy最新文献_第7页

CRISPR-AuNP: physicochemical optimization of a gold nanoparticle platform for cost-effective and modular non-viral gene editing in HSPCs CRISPR-AuNP：在HSPCs中进行成本效益和模块化非病毒基因编辑的金纳米颗粒平台的物理化学优化。

IF 4.5 3区医学

Gene Therapy Pub Date : 2026-01-14 DOI: 10.1038/s41434-025-00591-0

Karthikeya S. V. Gottimukkala, Daniel D. Lane, Rachel Cunningham, Haleema S. Malik, Youngseo Jwa, Molly E. Cassidy, Jack M. P. Castelli, Mark R. Enstrom, Katrina Poljakov, Grady Gastelum, Samuel H. Ho, Carlos Tassa, Jennifer E. Adair

{"title":"CRISPR-AuNP: physicochemical optimization of a gold nanoparticle platform for cost-effective and modular non-viral gene editing in HSPCs","authors":"Karthikeya S. V. Gottimukkala, Daniel D. Lane, Rachel Cunningham, Haleema S. Malik, Youngseo Jwa, Molly E. Cassidy, Jack M. P. Castelli, Mark R. Enstrom, Katrina Poljakov, Grady Gastelum, Samuel H. Ho, Carlos Tassa, Jennifer E. Adair","doi":"10.1038/s41434-025-00591-0","DOIUrl":"10.1038/s41434-025-00591-0","url":null,"abstract":"Efficient delivery of CRISPR ribonucleoproteins into primary hematopoietic stem and progenitor cells (HSPCs) is essential for durable gene editing therapies but remains challenging. Here, we advance a modular, benchtop-assembled gold-polymer hybrid nanoparticle (CRISPR-AuNP) platform that enables non-viral delivery of multiple CRISPR systems into HSPCs. Guided by a mechanistic understanding of Cas9’s interaction with gold surfaces, we engineered the formulation by conjugating pre-formed RNP-polymer complexes, assembled using thiolated polyethyleneimine-polyethylene glycol, to gold nanoparticles. This system achieved efficient editing in primary CD34+ HSPCs for Cas9, Cas12a, and Cas12a-M29-1 without compromising cell viability. Notably, the nanoformulation can be assembled in under 2 h in a PCR tube for less than $70/million HSPCs treated. This work establishes a scalable, cost-effective, and accessible gene editing system with the potential to democratize CRISPR applications in HSPC research and therapy.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 2","pages":"188-202"},"PeriodicalIF":4.5,"publicationDate":"2026-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.comhttps://www.nature.com/articles/s41434-025-00591-0.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145970431","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}

引用次数: 0

The co-delivery of Programmed Death 1 ligands enhances and prolongs rAAV-mediated gene expression in pre-immunized mice 程序性死亡1配体的共递送增强并延长了raav介导的基因在预免疫小鼠中的表达。

IF 4.5 3区医学

Gene Therapy Pub Date : 2026-01-09 DOI: 10.1038/s41434-025-00588-9

Piia Käyhty, Tiina Nieminen, Reetta A. E. Eriksson, Ritva Tumelius, Ahmed Tawfek, Svetlana Laidinen, Anna-Kaisa Ruotsalainen, Aubrey Bailey, Lionel Galibert, Hanna P. Lesch, Seppo Ylä-Herttuala, Kari J. Airenne

{"title":"The co-delivery of Programmed Death 1 ligands enhances and prolongs rAAV-mediated gene expression in pre-immunized mice","authors":"Piia Käyhty, Tiina Nieminen, Reetta A. E. Eriksson, Ritva Tumelius, Ahmed Tawfek, Svetlana Laidinen, Anna-Kaisa Ruotsalainen, Aubrey Bailey, Lionel Galibert, Hanna P. Lesch, Seppo Ylä-Herttuala, Kari J. Airenne","doi":"10.1038/s41434-025-00588-9","DOIUrl":"10.1038/s41434-025-00588-9","url":null,"abstract":"Immune responses against recombinant adeno-associated virus (rAAV) are one of the major obstacles in gene therapy. We investigated the potential of Programmed Death 1 ligands 1 and 2 (PD-L1/2) to protect AAV-transduced cells from immunological clearance. Ligand compatibility for co-delivery was first evaluated using two transgenes, VEGF-B186 and muSEAP, separated from PD-L1/2 by a self-cleaving P2A peptide. After proper cleavage and biological activity of the co-produced proteins were demonstrated in vitro, the effect of PD-L1/2 co-expression on muSEAP production and persistence was studied in naïve and vector pre-immunized mice. Vectors (rAAV6-muSEAP, rAAV6-muSEAP-PD-L1, or rAAV6-muSEAP-PD-L2) were injected into two sites of the gastrocnemius muscle at a total dose of 1×1010 vg. Co-delivery of PD-L1, particularly, significantly enhanced muSEAP secretion into the bloodstream up to 12 weeks despite elevated anti-AAV6 responses in pre-immunized mice. muSEAP secretion increased 33.3- and 31.4-fold with the co-delivery of PD-L1, while the increase was only 5.6- and 9.3-fold in the muSEAP control group at 5 and 12 weeks, respectively. Ligand-treated pre-immunized animals also had less T-cell infiltration into the treated muscle compared to naïve animals. In summary, co-delivery of PD-L1/2 alongside a transgene represents a promising strategy for achieving sustained gene expression in individuals pre-exposed to AAV.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 2","pages":"127-137"},"PeriodicalIF":4.5,"publicationDate":"2026-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.comhttps://www.nature.com/articles/s41434-025-00588-9.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145943229","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}

引用次数: 0

Evaluation of methods to increase the expression of cytokine-induced killer cell chemoattractant cytokines in pancreatic cancer 增加胰腺癌细胞因子诱导的杀伤细胞趋化因子表达方法的评价。

IF 4.5 3区医学

Gene Therapy Pub Date : 2026-01-09 DOI: 10.1038/s41434-025-00590-1

Maria Bunuales, Susana Inoges, Ascension Lopez-Diaz de Cerio, Sandra Hervas-Stubbs, Javier Rodriguez, Manuela Gonzalez-Aparicio, Patricia Jauregui, Ruben Hernandez-Alcoceba

{"title":"Evaluation of methods to increase the expression of cytokine-induced killer cell chemoattractant cytokines in pancreatic cancer","authors":"Maria Bunuales, Susana Inoges, Ascension Lopez-Diaz de Cerio, Sandra Hervas-Stubbs, Javier Rodriguez, Manuela Gonzalez-Aparicio, Patricia Jauregui, Ruben Hernandez-Alcoceba","doi":"10.1038/s41434-025-00590-1","DOIUrl":"10.1038/s41434-025-00590-1","url":null,"abstract":"Adoptive Cell Therapies based on cytokine-induced killer cells (CIKs) can address the heterogeneity of solid tumors due to their multiple mechanisms of cancer cell recognition. However, tumor trafficking is one of the main limitations. In this work, we describe that a high proportion of CIKs obtained from pancreatic ductal adenocarcinoma patients express the CXCR3 and CCR5 receptors, and they migrate towards their corresponding chemokines CXCL10 and CCL5 in vitro. Using an immune competent orthotopic PDAC mouse model, we have investigated the ability of different clinically compatible interventions to increase the expression of these chemokines. No significant elevation was obtained with chemotherapy (5-fluorouracil, irinotecan, oxaliplatin, paclitaxel, gemcitabine or temozolomide), tyrosine kinase inhibitors sorafenib and sunitinib, or the immunostimulatory agents polyinosinic:polycytidylic acid, Mycobacterium tuberculosis antigens and diphtheria/pertussis/tetanus vaccine. In contrast, CXCL10 and CCL5 expression was stimulated by local administration of an adenoviral vector equipped with a drug-inducible expression system for interleukin-12 (IL-12). Combination of the vector and CIKs obtained a strong antitumor effect in the PDAC model, although it was mainly due to vector-mediated recruitment of endogenous immune cells. We conclude that additional barriers beyond chemokine expression should be overcome in order to unleash the full potential of CIKs on solid tumors.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 3","pages":"275-283"},"PeriodicalIF":4.5,"publicationDate":"2026-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145943170","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

Metadata assessment of non-human primate studies of AAV9 uncovers potential tissue specific variation in expression efficiency 对非人类灵长类动物AAV9研究的元数据评估揭示了潜在的组织特异性表达效率差异。

IF 4.5 3区医学

Gene Therapy Pub Date : 2026-01-07 DOI: 10.1038/s41434-025-00589-8

Muhammad Shahrukh, Julianne R. Sweeney, Tony del Rio, Fatih Ozsolak

{"title":"Metadata assessment of non-human primate studies of AAV9 uncovers potential tissue specific variation in expression efficiency","authors":"Muhammad Shahrukh, Julianne R. Sweeney, Tony del Rio, Fatih Ozsolak","doi":"10.1038/s41434-025-00589-8","DOIUrl":"10.1038/s41434-025-00589-8","url":null,"abstract":"Adeno-associated virus (AAV)-based gene therapies have garnered significant attention and investment due to their clinical success and potential to address underlying causes of many diseases. AAV vectors provide effective delivery of therapeutic genetic material to disease-relevant tissues. When evaluating safety and efficacy of recombinant AAV vectors, biodistribution profiles play a critical role in novel therapy development. Herein, a biodistribution metadata analysis was performed on ten studies involving 51 cynomolgus macaques (Macaca fascicularis). The macaques received a self-complementary or single-stranded AAV9 vector containing chicken ß-actin (CBA) or cytomegalovirus (CMV173) promoters expressing fluorescent reporters or a human SMN1 gene. These studies covered various routes of administration (ROA) including intravenous (IV), intracisternal magna (ICM), and lumbar puncture intrathecal (IT) injection. Metadata analysis of AAV9 biodistribution showed relatively uniform vector genome delivery throughout spinal cord tissues over multiple timepoints and ROAs. Moreover, decreased expression efficiency of viral DNA in liver was observed regardless of the ROA, macaque age, or viral construct used. To understand this trend, epigenetic profiling of tissue-localized AAV9 vector genome DNA was performed. Experimental evidence supports partial silencing and repression of transgene expression in macaque liver. These findings point to plausible strategies to consider in preclinical development of AAV9 mediated gene therapies.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 3","pages":"266-274"},"PeriodicalIF":4.5,"publicationDate":"2026-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.comhttps://www.nature.com/articles/s41434-025-00589-8.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145917494","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}

引用次数: 0

Secondary malignancy of T-cell origin after CAR T-cell therapy: EMA’s conclusions from the evaluation of 38 suspected cases CAR - t细胞治疗后t细胞来源的继发性恶性肿瘤：EMA对38例疑似病例的评估结论

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-12-22 DOI: 10.1038/s41434-025-00586-x

Philipp Berg, Charlotte Bakker, Moritz Sander, Nicklas Hasselblad Lundstrøm, Karin Erneholm, Flora Musuamba Tshinanu, Olga Kholmanshikh, Filip Van Nuffel, Susanne Müller, Gabriele Ruppert-Seipp, Gabriele D. Maurer, Justina Januskiene, Maria Mantziri, Bianca Mulder, Frederika A. van Nimwegen, Daiana Vasilcanu, Ulla Wändel Liminga

{"title":"Secondary malignancy of T-cell origin after CAR T-cell therapy: EMA’s conclusions from the evaluation of 38 suspected cases","authors":"Philipp Berg, Charlotte Bakker, Moritz Sander, Nicklas Hasselblad Lundstrøm, Karin Erneholm, Flora Musuamba Tshinanu, Olga Kholmanshikh, Filip Van Nuffel, Susanne Müller, Gabriele Ruppert-Seipp, Gabriele D. Maurer, Justina Januskiene, Maria Mantziri, Bianca Mulder, Frederika A. van Nimwegen, Daiana Vasilcanu, Ulla Wändel Liminga","doi":"10.1038/s41434-025-00586-x","DOIUrl":"10.1038/s41434-025-00586-x","url":null,"abstract":"This article provides a regulatory perspective on secondary malignancy of T-cell origin as a rare adverse reaction to the currently marketed CD19- or BCMA-directed chimeric antigen receptor (CAR) T-cell therapies. To assess the risk, causality between reported suspected adverse reactions and CAR T-cell therapy was assessed applying the principles of the World Health Organization-Uppsala Monitoring Centre causality categories, alongside a review of scientific publications and data from registries/ databases. By 11 April 2024, 38 cases of T-cell malignancy after CAR T-cell therapy were reported in patients aged 29–80 years. In 19 patients, tumour samples were tested for the presence of CAR transgene, which was detected in seven cases. Most of the T-cell malignancies were diagnosed within 12 months of treatment (22/33; 67%). The reporting rate is approximately one case per 1000 patients treated. An overall causal relationship was established with at least a reasonable possibility. Regulatory measures included updates to the product information, risk management plan, and educational materials. An additional pharmacovigilance activity was requested from the marketing authorisation holders (MAHs) to strengthen the process of genetic testing of residual tumour samples. To further characterise this risk and understand underlying mechanisms, continued efforts from healthcare professionals, MAHs and regulators are essential. Well-documented case reports, including information on genetic testing of tumour samples, are considered crucial elements.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 3","pages":"260-265"},"PeriodicalIF":4.5,"publicationDate":"2025-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.comhttps://www.nature.com/articles/s41434-025-00586-x.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145809298","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}

引用次数: 0

Lipid nanoparticle mediated base editing of the Q344X rhodopsin mutation associated with retinitis pigmentosa. 脂质纳米颗粒介导的与视网膜色素变性相关的Q344X视紫红质突变的碱基编辑

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-12-10 DOI: 10.1038/s41434-025-00584-z

Victoria A C Palmgren, Miffy Hok Yan Cheng, Yao Zhang, Tiffany Carlaw, Tessa Morin, Jerry Leung, Colin Jd Ross, Pieter R Cullis, Robert S Molday

{"title":"Lipid nanoparticle mediated base editing of the Q344X rhodopsin mutation associated with retinitis pigmentosa.","authors":"Victoria A C Palmgren, Miffy Hok Yan Cheng, Yao Zhang, Tiffany Carlaw, Tessa Morin, Jerry Leung, Colin Jd Ross, Pieter R Cullis, Robert S Molday","doi":"10.1038/s41434-025-00584-z","DOIUrl":"https://doi.org/10.1038/s41434-025-00584-z","url":null,"abstract":"Retinitis pigmentosa (RP) associated with mutations in the rhodopsin gene (RHO) is a significant cause of blindness. Here we report on the application of adenine base editing of the c.1030C>T (p.Q344X) RHO mutation linked to RP. Using a fluorescence reporter cell system, we optimized editing by exploring base editors, sgRNA, and delivery methods. Flow cytometry, western blotting, and immunofluorescence microscopy confirmed the restoration of full-length rhodopsin after editing. DNA sequencing verified editing at the target nucleotide and the absence of bystander edits within the editing window. Polyethylenimine cationic polymer transfection of cells with a plasmid containing the NG-ABE8e adenine base editor and A6 guide RNA that placed the targeted adenine in position 6 of the editing window resulted in 31.0% gDNA sequence correction and 26.3% rhodopsin protein correction as determined by flow cytometry. Purified NG-ABE8e protein complexed with A6-sgRNA showed 32.2% gDNA editing and 44.5% rhodopsin correction. Plasmid NG-ABE8e and A6-sgRNA co-encapsulated into lipid nanoparticles (LNPs) and transfected into the reporter cell system resulted in the highest editing (42.6% gDNA editing and 65.9% rhodopsin correction). These results demonstrate the successful correction of the c.1030C>T RHO mutation and provide the foundation for base editing as a treatment for RP.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145722583","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

Intranasal versus intravenous AAV delivery: A comparative analysis of brain-targeting efficiency and peripheral exposure in mice 鼻内与静脉内AAV递送：小鼠脑靶向效率和外周暴露的比较分析。

IF 5.9 3区医学

Gene Therapy Pub Date : 2025-12-08 DOI: 10.1038/s41434-025-00585-y

Chinwendu Chukwu, Jinyun Yuan, Hong Chen

{"title":"Intranasal versus intravenous AAV delivery: A comparative analysis of brain-targeting efficiency and peripheral exposure in mice","authors":"Chinwendu Chukwu, Jinyun Yuan, Hong Chen","doi":"10.1038/s41434-025-00585-y","DOIUrl":"10.1038/s41434-025-00585-y","url":null,"abstract":"Adeno-associated viruses (AAVs) hold significant promise for gene therapy targeting the central nervous system (CNS). However, current delivery methods are either invasive or cause significant systemic exposure. Intranasal (IN) delivery presents a promising noninvasive alternative for direct CNS targeting, though its efficacy in delivering AAVs to the brain has seldom been explored. Here, we quantitatively assessed AAV transduction in the brain and peripheral organs of Swiss, BALB/c, and C57BL/6 J mice following IN administration, using intravenous (IV) injection as a benchmark for comparison. Our findings revealed that IN administration of the AAV9 vector achieved approximately 15% of the transduction efficiency and 9% of the gene expression levels observed with IV delivery. Importantly, IN delivery significantly reduced systemic exposure to most major peripheral organs by up to 1.34 × 104-fold compared to IV injection. The ratios of gene transduction between the brain and various peripheral tissues were calculated, revealing that for key organs such as the liver, stomach, kidney, and spleen, IN delivery achieved higher brain-to-peripheral transduction ratios than IV delivery. These findings underscore the potential of IN delivery for noninvasive brain-targeted gene delivery with significant reductions in peripheral exposure.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 4","pages":"486-494"},"PeriodicalIF":5.9,"publicationDate":"2025-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12851798/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145707868","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}

引用次数: 0

Precision rewriting of muscle genetics: therapeutic horizons of base and prime editing in skeletal muscle disorders 肌肉遗传学的精确重写：骨骼肌疾病的碱基和起始编辑的治疗视野。

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-12-04 DOI: 10.1038/s41434-025-00574-1

Selin Saydam, Pervin Dinçer

{"title":"Precision rewriting of muscle genetics: therapeutic horizons of base and prime editing in skeletal muscle disorders","authors":"Selin Saydam, Pervin Dinçer","doi":"10.1038/s41434-025-00574-1","DOIUrl":"10.1038/s41434-025-00574-1","url":null,"abstract":"Base Editing (BE) and Prime Editing (PE), novel precision tools of the CRISPR/Cas toolbox, have emerged as transformative technologies that enable highly specific genetic modifications. Their compatibility with post-mitotic cell types makes them invaluable for treating genetic skeletal muscle disorders. Despite their severity and progressive nature, monogenic muscle diseases remain without definitive treatments. They are caused by diverse mutations in critical muscle proteins, for which gene editing offers a promising therapeutic avenue. However, traditional CRISPR/Cas9 applications face challenges such as genotoxicity and inefficiency in post-mitotic tissues. BE and PE technologies overcome these limitations by enabling safe and efficient modifications without causing double-strand breaks or requiring homology-directed repair. Their therapeutic potential comes from two key features: their ability to work in non-dividing cells such as myotubes and cardiomyocytes, and their capacity to target a broad range of mutations found in genetic muscle diseases. In this review, we explore mechanisms of BE and PE and summarize their current applications in monogenic skeletal muscle disorders. We discuss the challenges of in vivo application in skeletal muscle and highlight innovations to bypass them. Collectively, both systems offer flexible precision solutions with immense potential for mutation-specific and personalized gene therapy approaches for monogenic skeletal muscle disorders.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 3","pages":"249-259"},"PeriodicalIF":4.5,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145677185","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

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 5.9 3区医学

Gene Therapy Pub Date : 2025-12-01 Epub Date: 2025-09-28 DOI: 10.1038/s41434-025-00566-1

Stefano Cattaneo, Barbara Bettegazzi, Selene Ingusci, Gianluca Verlengia, Anna Sofia Tascini, Silvia Zucchini, 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, Anna Sofia Tascini, Silvia Zucchini, Franca Codazzi, Marco J Morelli, Marco Marzulli, Joseph C Glorioso, Michele Simonato","doi":"10.1038/s41434-025-00566-1","DOIUrl":"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":"581-593"},"PeriodicalIF":5.9,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12714583/pdf/","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":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Dystrophin/mini-dystrophin expression analysis by immunoaffinity liquid chromatography-tandem mass spectrometry after gene therapy for DMD. 免疫亲和液相色谱-串联质谱法分析DMD基因治疗后肌营养不良蛋白/微肌营养不良蛋白的表达。

IF 4.5 3区医学

Gene Therapy Pub Date : 2025-12-01 Epub Date: 2025-08-02 DOI: 10.1038/s41434-025-00554-5

Jason Walsh, Joe Palandra, Nicole Duriga, David Beidler, Avery McIntosh, Michael Binks, Hendrik Neubert

{"title":"Dystrophin/mini-dystrophin expression analysis by immunoaffinity liquid chromatography-tandem mass spectrometry after gene therapy for DMD.","authors":"Jason Walsh, Joe Palandra, Nicole Duriga, David Beidler, Avery McIntosh, Michael Binks, Hendrik Neubert","doi":"10.1038/s41434-025-00554-5","DOIUrl":"10.1038/s41434-025-00554-5","url":null,"abstract":"Adeno-associated virus (AAV)-based gene replacement therapies in Duchenne muscular dystrophy (DMD) aim to restore dystrophin function via the introduction of micro- or mini-dystrophins. We report dystrophin and mini-dystrophin concentrations generated by immunoaffinity liquid chromatography-tandem mass spectrometry (IA-LC-MS/MS) in skeletal muscle biopsies from ambulatory participants with DMD in a phase 1b study of fordadistrogene movaparvovec, an AAV9-based gene replacement construct. The assay performed robustly for 26 months, as demonstrated by limited variability in calibration standards for peptides LLQV (dystrophin and mini-dystrophin) and LEMP (mini-dystrophin only), quality control samples consisting of spiked mini-dystrophin in DMD skeletal muscle lysate, as well as unspiked, pooled, non-dystrophic skeletal muscle lysate (normal pool). Average values for LLQV in the normal pool tested as part of clinical sample and long-term stability runs were similar to validated values. Biopsy samples showed minor or absent LLQV and absent LEMP signals pre-treatment with fordadistrogene movaparvovec infusion, but signals substantially increased at Days 60 and 360, on average. There was strong concordance in LEMP and LLQV expression change between Days 60 and 360 (R2 = 0.91; p < 0.001). IA-LC-MS/MS enables reproducible, stable, and reliable quantification of dystrophin/mini-dystrophin following fordadistrogene movaparvovec infusion. ClinicalTrials.gov identifier: NCT03362502.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":"573-580"},"PeriodicalIF":4.5,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12714582/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144768611","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}

引用次数: 0