Gene Therapy最新文献

{"title":"FMR1 gene therapy restores translationally relevant phenotypes in a mouse model for fragile X syndrome","authors":"Richard K. Lacher, Kari Henson, Lindsay N. Wathen, Caitlin Jones, Tiffany Arnold, McKenzie R. Rice, Heather M. Carles, Angela R. White, Elizabeth Ramsuchit, Darren Murrey, Rebecca Raig, Austen Fisher, Kaitlin Bucher, Grace C. Westerkamp, Adam L. Fritz, Brooke M. Gollaway, Sebastian Piloto, David Dismuke, J Elliott Robinson, Michael T. Williams, Charles V. Vorhees, Erandi K. De Silva, Durgesh Tiwari, Craig A. Erickson, Ernest V. Pedapati, Christina Gross","doi":"10.1038/s41434-026-00630-4","DOIUrl":"10.1038/s41434-026-00630-4","url":null,"abstract":"Fragile X Syndrome (FXS) is the most common inherited form of intellectual disability. It is caused by a trinucleotide expansion in the 5’ UTR of the Fragile X messenger ribonucleoprotein 1 (FMR1) gene leading to loss of expression of Fragile X messenger ribonucleoprotein (FMRP). There is currently no cure for FXS. We developed an FMR1 gene therapy based on an adeno-associated viral vector designed with strong translational potential for future clinical testing. The viral vector was tested in Fmr1 knockout mice using two translationally relevant delivery routes and ages corresponding to in utero, toddler, and adolescent ages in humans. Functional studies showed that the FMR1 gene therapy improved select translational FXS phenotypes spanning three critical domains: sensory hyperexcitability, adaptation to change, and altered brain activity. Expression after intracerebroventricular injection was most prominent in the forebrain, whereas intravenous delivery predominantly led to expression across midbrain and brainstem, suggesting that a dual route may be needed to achieve full brain coverage. Biodistribution analyses further suggested that FMRP expression must be titrated carefully for optimal rescue. In summary, we show that FMR1 gene therapy using delivery routes and vehicles approved for clinical use improves core phenotypes in a mouse model for FXS.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 5","pages":"515-531"},"PeriodicalIF":5.9,"publicationDate":"2026-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.comhttps://www.nature.com/articles/s41434-026-00630-4.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148420549","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":"Improving the precision of AAV lung gene therapy for SP-B deficiency using computationally derived lung-specific promoters","authors":"Nicole Zielinska, Erin. L. Howard, Brenna A. Y. Stevens, Melanie M. Goens, Elena S. B. Campbell, Madison E. Hughes, Yanlong Pei, Liqun Xu, Adithya Achuthan, Arul Vadivel, Arnold Apostol, Xiaolin He, Dinghai Zheng, D. Benjamin Gordon, Raja R. Srinivas, Alec A. K. Nielsen, Jeff L. Caswell, Luis G. Arroyo, Darren A. Yuen, Bernard Thebaud, Sarah K. Wootton","doi":"10.1038/s41434-026-00629-x","DOIUrl":"10.1038/s41434-026-00629-x","url":null,"abstract":"Recombinant adeno-associated virus (rAAV) platforms have achieved significant success in clinical gene therapy; however, many still rely on ubiquitous promoters. This robust and widespread transgene expression can cause off-target effects, immune activation, and systemic toxicity, limiting their suitability for diseases requiring tissue-specific expression, such as surfactant protein B (SP-B) deficiency. Here, we aimed to improve the precision of AAV-lung gene therapy by evaluating computationally predicted lung-specific promoters with AAV6.2FF, a capsid with strong lung tropism. Promoter strength and specificity were assessed following administration of AAV6.2FF encoding the human placental alkaline phosphatase (AP) reporter gene in mice. Cross-species activity was evaluated in precision-cut lung slices (PCLS) from ferrets and pigs. We identified promoter 5979 as lung-specific in mice, outperforming the ubiquitous CASI promoter (comprised of the cytomegalovirus enhancer, chicken β-actin promoter, and ubiquitin C regulatory elements) in transgene expression and specificity, independent of AAV capsid or route of administration. In a conditional SP-B knockout model, AAV6.2FF-5979-hSPB extended survival in SP-B-deficient mice and outperformed its CASI-driven counterpart. Promoter 5979 exhibited the highest activity among the four synthetic promoters in ferret PCLS, but demonstrated limited activity in pig PCLS, underscoring species-specific differences. This study demonstrates the therapeutic value of tissue-specific promoters in targeted gene therapy while emphasizing the importance of refining algorithmic prediction platforms to ensure reliable cross-species and clinical performance.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 5","pages":"571-585"},"PeriodicalIF":5.9,"publicationDate":"2026-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148360418","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":"Recent advancements in improving cross-species applicability of bioengineered AAV capsids","authors":"Haolai Pan, Nianci Li, Jieyu Qi, Renjie Chai","doi":"10.1038/s41434-026-00628-y","DOIUrl":"10.1038/s41434-026-00628-y","url":null,"abstract":"Adeno-associated virus (AAV) is widely accepted as a delivery vector for in vivo gene therapy due to its relatively low immunogenicity, minimal toxicity, sustained efficacy, and broad tropism. However, its unpredictable cross-species applicability remains a troublesome hurdle for broader clinical applications. Thus, designing novel AAV capsids with enhanced cross-species applicability is urgently needed. In this review, we present AAV bioengineering methods, including rational design, directed evolution, and artificial intelligence–based design, with the goal of creating novel AAV variants that are translatable to humans. Using representative examples, we also evaluate how each method addresses key species-dependent barriers—receptor usage, intracellular trafficking, immune recognition, and toxicity—that critically determine cross-species translatability.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 5","pages":"635-643"},"PeriodicalIF":5.9,"publicationDate":"2026-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148351755","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":"Assessment of F/HN-pseudotyped lentiviral vector following intravenous delivery to mice","authors":"Robyn V. Bell, Nikhil B. Faulkner, Anthony Sinadinos, Cuixiang Meng, Emily Castells, Mariana A. Viegas, Stephen C. Hyde, Deborah R. Gill, Eric WFW Alton, Uta Griesenbach","doi":"10.1038/s41434-026-00623-3","DOIUrl":"10.1038/s41434-026-00623-3","url":null,"abstract":"In pursuit of a gene transfer agent with efficient pulmonary transduction, the UK Respiratory Gene Therapy Consortium has developed a lentiviral vector pseudotyped with the envelope proteins, F and HN from Sendai virus (rSIV.F/HN). In contrast to other viral vectors, pulmonary rSIV.F/HN delivery achieves sustained gene expression ( ~ 2 years in mice) in the lungs and systemic circulation following a single dose. Here, we investigate the application of the rSIV.F/HN vector-platform for wider indications, including systemic disorders that require serum expression of therapeutic proteins. To assess the potential for rSIV.F/HN to produce systemic proteins, intravenous vector delivery was characterised and compared against intrapulmonary administration, achieved via ‘nasal sniffing’. Both delivery routes achieved sustained (at least 1 year) systemic expression of the secreted reporter protein Gaussia luciferase. Systemic rSIV.F/HN delivery resulted in widespread protein expression across multiple organs, accompanied by the generation of significant anti-vector neutralising antibodies limiting vector readministration. Conversely, localised airway transduction was observed following pulmonary administration, which we have previously shown is not an impediment to efficient vector readministration. These data support intrapulmonary rSIV.F/HN delivery for systemic protein production, with sustained high-level transgene expression and feasible readministration.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 5","pages":"586-595"},"PeriodicalIF":5.9,"publicationDate":"2026-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.comhttps://www.nature.com/articles/s41434-026-00623-3.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148277321","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":"Applications of genome editing technologies in the treatment of human diseases.","authors":"Jamal Alshorman, Mohammad Javad Mehran, Kingsley Miyanda Tembo, Niusha Mostafavi, Mansoor Bolideei, Yongping Wang","doi":"10.1038/s41434-026-00627-z","DOIUrl":"https://doi.org/10.1038/s41434-026-00627-z","url":null,"abstract":"<p><p>Genome editing has progressed from a laboratory capability for targeted DNA manipulation to a clinically relevant strategy for correcting, silencing, or regulating genes implicated in human disease. In this Review, we synthesize the mechanisms, capabilities, and constraints of the principal programmable platforms-zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR-Cas systems-and highlight how base editors, prime editors, and epigenetic editors expand the range of achievable outcomes beyond double-strand break-dependent repair to precise nucleotide substitutions, small insertions/deletions, and transcriptional modulation. We compare genome-editing cargo formats, including plasmid DNA, viral-vector DNA, mRNA, guide RNA, and ribonucleoprotein complexes, together with the delivery modalities used to transport them, including AAV, adenoviral and herpesviral vectors, lipid nanoparticles (LNPs), electroporation, and virus-like particles. We then consolidate key biomedical applications enabled by these technologies, spanning endogenous gene tagging, high-throughput functional variant screening, molecular recording, and the generation of genetically faithful disease models. Across oncology, respiratory, hematologic, cardiovascular, metabolic, neurodegenerative, viral, ocular, and immune disorders, genome editing is advancing both ex vivo and in vivo interventions, including engineered cellular immunotherapies, hematopoietic stem and progenitor cell editing for hemoglobinopathies, and emerging liver-directed programs for lipid and coagulation targets. Finally, we discuss priorities for broad clinical implementation: improving editing fidelity and PAM flexibility, increasing performance in non-dividing cells, enabling tissue-selective delivery to difficult organs (for example, lung and central nervous system), and addressing manufacturing scalability, long-term monitoring, and equitable global access.</p>","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2026-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148258008","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":"High resolution ES-DMA quantifies AAV capsid DNA content by electrical mobility to mass correlation","authors":"Preston Dennett,&nbsp;Lohra M. Young,&nbsp;Benjamin E. Draper,&nbsp;Jerome Schmitt,&nbsp;Martin Jarrold,&nbsp;Luis Javier Perez-Lorenzo","doi":"10.1038/s41434-026-00626-0","DOIUrl":"10.1038/s41434-026-00626-0","url":null,"abstract":"Recombinant Adeno-Associated Virus (rAAV) is the leading viral vector platform for gene therapy. A persistent challenge in rAAV manufacturing and quality control is accurate assessment of the DNA content of purified viral capsids. Empty particles, as well as ‘partial’ and over-filled species commonly contaminate even highly purified rAAV preparations. These impurities contribute to high production costs and reduced therapeutic efficacy, resulting in higher required doses and increased risks to patient health. Here we present high-resolution electrospray differential mobility analysis (ES-DMA) as a novel benchtop analytical modality for rAAV vector characterization. Sampling microliter volumes of analyte, this method achieves angstrom-scale particle sizing which we demonstrate is sufficient to resolve DNA encapsidation by rAAVs via differential electrical mobility. We benchmark this approach against charge detection mass spectrometry (CD-MS), an emerging gold standard for rAAV analytics, and identify a robust near-linear correlation between electrical mobility and mass of aerosolized rAAV particles. This relationship enables rapid identification and relative quantification of empty, full, and partial / over-filled rAAV capsids by ES-DMA.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 5","pages":"609-615"},"PeriodicalIF":5.9,"publicationDate":"2026-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.comhttps://www.nature.com/articles/s41434-026-00626-0.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148163052","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":"AAV8-mediated mouse/human PROC expression rescues thrombophilia in hereditary protein C-deficient mice.","authors":"Tingting Wu, Yanyi Tao, Hui Lu, Yaohua Cai, Yunqing Xia, Tingting Liu, Li Wang, Zhipeng Cheng, Yu Hu, Javier Corral, Liang V Tang","doi":"10.1038/s41434-026-00624-2","DOIUrl":"https://doi.org/10.1038/s41434-026-00624-2","url":null,"abstract":"<p><p>Hereditary protein C (PC) deficiency, which is caused by PROC gene mutations, increases the risk of venous thromboembolism and offers limited treatment options. In this study, we developed adeno-associated virus serotype 8 (AAV8) vectors carrying either murine PC (AAV8-mPROC) or human PC (AAV8-hPROC) transgenes. These vectors were delivered through tail vein injection into PROC knockout mice. The highest dose of AAV8-mPROC (6.00E + 12 vg/kg) resulted in PC activity and antigen levels reaching 200.7% and 190.1%, respectively, which were maintained at 171.8% and 165.2%, respectively, by week 48. Similarly, the highest dose of AAV8-hPROC (8.00E + 12 vg/kg) resulted in 295.4% PC activity and 3.72 μg/ml human protein C antigen, which were maintained at 195.1% and 1.13 μg/ml, respectively, by week 48. In the vein thrombosis model, AAV8-mPROC and AAV8-hPROC significantly reduced the thrombus weight from 12.11 ± 3.39 mg to 7.19 ± 2.28 mg and 6.81 ± 2.28 mg, respectively. In the pulmonary embolism model, the proportion of embolized vessels decreased from 88.53% to approximately 60.62% in the AAV8-mPROC group and 62.33% in the AAV8-hPROC group. Our study has established a preclinical foundation for the safe and effective application of AAV vector-based gene therapy in treating inherited PC deficiency.</p>","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148143093","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":"Environmental and developmental factors shape anti-AAV immunity in pigs.","authors":"Godwin I Iroanya, Ilangovan Raju, Chandra Boosani, Pradeep N Subramanyam, Addison K Byrne, Kevin D Wells, Jonathan A Green","doi":"10.1038/s41434-026-00625-1","DOIUrl":"10.1038/s41434-026-00625-1","url":null,"abstract":"<p><p>Use of adeno-associated virus (AAV) vectors has revolutionized in vivo gene therapy, but the presence of pre-existing neutralizing antibodies remains a major barrier that can hinder clinical application. While large-animal models such as non-human primates have been used to study anti-AAV immunity, their high cost and limited accessibility present challenges for studying the impact of AAV immunity on AAV-based therapies. Here, we evaluate pigs as an immunologically relevant large-animal model for investigating humoral barriers to AAV-based gene therapy. Using ELISA-based profiling across 11 AAV serotypes, we detected immunoglobulin G (IgG) antibodies against AAV capsids in pigs as early as two weeks of age, with titers increasing with age and displaying serotype-specific dynamics. Animals maintained in standard housing displayed greater inter-individual variations and broader serotype-specific reactivities. Functional assays demonstrated that these antibodies neutralized AAV particles in a dose- and serotype-dependent manner, with IgG depletion restoring transduction in vitro. Sequence analysis indicated that capsid identity can partially predict cross-reactive binding, but only under controlled conditions. These findings establish pigs as a tunable model capable of recapitulating age- and environment-dependent features of anti-AAV immunity, providing a platform for studying humoral barriers and evaluating immune evasion strategies in AAV-based gene therapy.</p>","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":""},"PeriodicalIF":5.9,"publicationDate":"2026-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13333265/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148054160","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":"Characterization of CAR-T cell factors that contribute to myeloid cell activation","authors":"Supreet Khanal,&nbsp;Md Kamal Hossain,&nbsp;Joseph Fischer,&nbsp;Santosh Panthi,&nbsp;Alan Baer,&nbsp;Nirjal Bhattarai","doi":"10.1038/s41434-026-00620-6","DOIUrl":"10.1038/s41434-026-00620-6","url":null,"abstract":"Chimeric antigen receptor (CAR) T cell therapies have shown remarkable success in the treatment of hematologic cancers; however, their use is often accompanied by inflammatory toxicities, including cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). These toxicities, ranging from mild to life-threatening, are partly driven by bystander myeloid cell activation (BMCA) and the subsequent release of pro-inflammatory cytokines such as IL-6 and IL-1β. Although previous studies have described the individual contributions of GM-CSF, IFN-γ, and TNFα secreted by CAR-T cells, a comprehensive characterization of CAR-T-derived inflammatory factors has been lacking. In this study, we characterized the soluble factors secreted by activated CAR-T cells derived from human peripheral blood and assessed their role in BMCA. Comparative cytokine analyses across human T cell subsets, including CAR-T cells, identified multiple candidates involved in BMCA. Antibody-mediated neutralization confirmed that four factors, GM-CSF, IFN-γ, TNFα, and GPIbα, play dominant roles in driving BMCA. Furthermore, siRNA-mediated knockdown of these factors in CAR-T cells significantly reduced BMCA without impairing their anti-tumor activity. These findings are consistent with prior reports on the inflammatory roles of GM-CSF, IFN-γ, and TNFα during CAR-T cell therapy and, importantly, identify GPIbα as a previously unrecognized contributor to CAR-T-associated inflammatory toxicities. Targeting these factors through antibody blockade or genetic modification may represent a promising strategy to mitigate inflammatory toxicities and improve the safety of CAR-T cell therapies.","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":"33 5","pages":"616-627"},"PeriodicalIF":5.9,"publicationDate":"2026-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.nature.comhttps://www.nature.com/articles/s41434-026-00620-6.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148029434","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":"DOCTER: a genetically encoded switchable protein module for ERα-mediated transcriptional regulation.","authors":"Jinmin Wang, Jingfang Liu, Dandan Peng, Yuhao Ji, Ke Huang, Junfen Fu, Yingke Xu","doi":"10.1038/s41434-026-00622-4","DOIUrl":"https://doi.org/10.1038/s41434-026-00622-4","url":null,"abstract":"<p><p>The inhibition of estrogen receptor (ER)-mediated genomic signaling in ER-positive cancer cells has long been a primary focus of therapeutic strategies. Here, we introduce a switchable competitive inhibition system for ERα-mediated transcriptional regulation, termed DOCTER (Drug-induced On-Off Competitor for Transcription mediated by ERα). DOCTER integrates the Tet-On induced Cre-loxP recombination system to enable precise, reversible on/off switching. We demonstrate that DOCTER effectively inhibits ERα-mediated transcriptional regulation in breast cancer cells, modulating both exogenous and endogenous gene expression, and remains effective in cells harboring ERα ligand-binding domain (LBD) mutations. Upon drug induction, DOCTER exhibits controllable and reversible inhibition. To visualize these dynamic switching events in real time, we developed a multi-color fluorescent reporting system that enables monitoring of complete DOCTER switch-off within 24 hours. Our study provides a novel approach for time-specific transcriptional regulation and offers broad potential for applications in genetic research and therapeutic development.</p>","PeriodicalId":12699,"journal":{"name":"Gene Therapy","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2026-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148029444","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}