Human gene therapy最新文献

{"title":"Novel VSV-G Variants with Enhanced Blinding for Targeted Delivery of Lentiviral Vectors.","authors":"Felix L Warnecke, Moritz Ertelt, Anjali Shrivastava, Jennifer Harre, Athanasia Warnecke, Angela E Araujo, Hildegard Büning, Michael Morgan, Clara T Schoeder, Juliane W Schott, Axel Schambach","doi":"10.1177/10430342261466823","DOIUrl":"10.1177/10430342261466823","url":null,"abstract":"<p><p>Targeted <i>in vivo</i> transduction, entailing direct administration of viral vector preparations to patients, is the next big step in gene therapy. To redirect lentiviral vector (LV) particles selectively to desired cell type(s), different glycoproteins have been engineered to alter their tropism, including the glycoprotein G from Vesicular Stomatitis Virus (VSV-G) as the most commonly employed tropism-defining protein for LV particles. For detargeting from its natural receptor, the low-density lipoprotein receptor (LDLR), a VSV-G variant with two blinding substitutions, K47Q and R354A, is commonly used (VSV-G.pub). We provide first evidence for insufficient blinding of VSV-G.pub in human and murine cell lines and primary cells. <i>In silico</i> modeling pointed toward only slightly reduced LDLR binding affinity of VSV-G.pub. To lower the affinity further, we generated three novel VSV-G variants based on charge-reversing substitutions in two, four, or six key residues. These variants achieved a more stringent blinding compared with VSV-G.pub in the tested cell lines and primary cells. Codisplay of a CD4 binder enabled lentiviral particles pseudotyped with the novel variants to selectively transduce CD4-expressing cells. In summary, we present improved VSV-G variants with a better on/off-target ratio as attractive tools for <i>in vivo</i> gene therapy applications.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"770-787"},"PeriodicalIF":4.6,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148663781","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":"Helper-Dependent Adenoviral Vectors: Recent Advances and Emerging Applications.","authors":"Montaha Alshawabkeh, Eric Ehrke-Schulz, Wenli Zhang, Anja Ehrhardt","doi":"10.1177/10430342261460742","DOIUrl":"10.1177/10430342261460742","url":null,"abstract":"<p><p>Helper-dependent adenoviral vectors (HDAdVs), which lack all viral coding sequences, enable the delivery of up to 35 kilobases (kb) of therapeutic DNA into target cells. This large packaging capacity facilitates the transfer of complex, large, or even multiple transgenes. Despite the technical complexity of their production, HDAdVs have been extensively evaluated in numerous preclinical studies, and several clinical trials have recently been initiated. This review provides a comprehensive, state-of-the-art overview of recent advances in the HDAdVs technology. It summarizes key strategies for vector development, including capsid engineering and improvements in production methodologies. Furthermore, it reviews preclinical <i>in vivo</i> studies with a focus on vector design, target indications, and therapeutic outcomes. Most preclinical applications have focused on targeting the liver and hematopoietic stem cells; however, additional organ systems and vaccination approaches based on HDAdVs are also being actively explored. In addition to preclinical progress, three recently initiated clinical trials using HDAdVs targeting knee osteoarthritis, chronic granulomatous disease, and solid tumors are highlighted. Nevertheless, several challenges must be addressed to enable broader clinical translation of HDAdVs. These include the complexity of large-scale manufacturing, achieving efficient and specific vector targeting, and overcoming host immune responses. In conclusion, HDAdV-based vector systems hold considerable promise for the treatment of a wide range of diseases. Continued technological advancements, including the development of next-generation adenoviral platforms and the integration of precision gene therapy approaches, are likely to further enhance their potential as versatile biotherapeutic tools.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"670-690"},"PeriodicalIF":4.6,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148648523","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":"Metagenomic Reconstruction of Adeno-Associated Virus Genomes.","authors":"Olena Maiakovska, Lukas Link, Margarita Zayas, Yumi Sano, Teng Wei Koay, Kleopatra Rapti, Jixin Liu, Felix Bubeck, Man Xu, Conradin Baumgartl, Ellen Wiedtke, Chiara Krämer, Lena Naber, Dirk Grimm","doi":"10.1177/10430342261419534","DOIUrl":"10.1177/10430342261419534","url":null,"abstract":"<p><p>The exponential expansion of metagenomic data obtained through high-throughput sequencing technologies has surpassed the petabyte-scale threshold, resulting in an unprecedented abundance of data now enabling the <i>in silico</i> discovery of previously unknown viral and bacterial species. Here, we demonstrate the power and promise of mining sequencing data to uncover natural adeno-associated virus (AAV) cap(sid) genes, with the synergistic aims to expand our repertoire of templates for vector development and to enhance our understanding of the AAV space and of virus evolution across species. Specifically, we harnessed the Serratus Explorer to identify 29 AAV variant genomes from publicly accessible raw metagenomic data generated from birds, nonhuman primates, or human samples, of which 16 were classified as high-quality based on the high coverage of their <i>cap</i> region. To this end, we devised a comprehensive computational pipeline comprising (i) reference candidate selection, (ii) prealigned data acquisition, (iii) variant calling and frequency estimation, (iv) consensus calling, (v) variant resolution, (vi) phylogenetic analysis, and (vii) protein structure analysis steps. Eight representative <i>cap</i> genes from four different host organism species were synthesized and used to produce so-called metAAV vectors, which exhibited intriguing and biomedically relevant properties including partial escape from neutralizing anti-AAV antibodies and muscle tropism combined with robust liver detargeting in systemically injected mice. We concurrently pursued a conventional, reference-independent metagenome-based genome assembly, which also successfully reconstructed AAV <i>cap</i> genes but solely for abundant variants. Together with the fact that this traditional reference-independent method necessitates substantial computational resources and misses to accurately resolve multiple closely related variants, this highlights the assets and superiority of our original consensus-based reconstruction pipeline for fundamental virus research and for future gene therapy vector bioengineering.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"713-732"},"PeriodicalIF":4.6,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146179452","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":"Lilly, Ascidian Launch Up-to-$1.9B RNA Exon Editor Collaboration.","authors":"Alex Philippidis","doi":"10.1177/10430342261465184","DOIUrl":"10.1177/10430342261465184","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"634-638"},"PeriodicalIF":4.6,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148368167","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":"Genome Editing for Familial Hemophagocytic Lymphohistiocytosis: Design Principles, Challenges, and Translational Perspectives.","authors":"Ghazale Minaiyan, Clotilde Aussel, Sandra Ammann, Toni Cathomen","doi":"10.1177/10430342261465555","DOIUrl":"10.1177/10430342261465555","url":null,"abstract":"<p><p>Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening hyperinflammatory syndrome caused by genetic defects in cytotoxic lymphocyte function. Current therapies can control disease activity, but transplantation of allogeneic hematopoietic stem and progenitor cells (HSPCs) remains the only curative option and is associated with substantial risks. These limitations have accelerated development of genome editing approaches enabling precise correction of disease-causing mutations in autologous cells. Familial HLH (FHL) represents a compelling target for genome editing, but successful and safe clinical translation has remained challenging. Preclinical studies demonstrate that targeted editing of key genes, such as <i>PRF1</i> and <i>UNC13D</i>, can restore cytotoxic function in HSPCs and T cells. Translation to the clinic, however, depends on multiple factors, including the choice of target cell population, the level of functional correction required, and gene-specific constraints such as locus complexity and regulation of gene expression. In this review, we synthesize current progress in genome editing for FHL and highlight critical biological and technical barriers to clinical implementation. We propose a conceptual framework for designing genome editing strategies tailored to FHL, emphasizing the alignment of editing platform, gene architecture, and cellular context to enable effective and clinically translatable therapies.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"702-712"},"PeriodicalIF":4.6,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148368242","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":"Biodistribution of AAV-TT and AAV9 in the Nonhuman Primate Brain.","authors":"Alberto J Rico, Nelly Jolinon, Teresa Torre-Muruzabal, R Michael Linden, Els Henckaerts, José L Lanciego","doi":"10.1177/10430342261427733","DOIUrl":"10.1177/10430342261427733","url":null,"abstract":"<p><p>The adeno-associated virus (AAV) capsid variant known as \"true type\" (AAV-TT) is a capsid with enhanced central nervous system tropism and efficient retrograde transport across brain circuits. This property enables AAV-TT to transduce neurons in multiple regions projecting to the site of injection, as previously shown in rodent and sheep models. Such spreading offers the potential for broad therapeutic vector distribution to functionally connected brain areas via a single intraparenchymal administration. To assess the clinical potential of AAV-TT, we delivered a green fluorescent protein (GFP)-expressing AAV-TT vector into the left commissural putamen of two young adult <i>Macaca fascicularis</i>. For comparison, two additional animals received AAV9-GFP. Four weeks post-injection, animals were euthanized, and whole-brain transduction patterns were analyzed using a dedicated Aiforia^® deep learning-based image analysis algorithm to quantify GFP+ neurons. We observed GFP expression in structures known to innervate the injection site, consistent with retrograde transport. Both AAV-TT and AAV9 showed similar overall distribution patterns, but AAV-TT demonstrated modestly enhanced neuronal transduction efficiency compared with AAV9. This study provides the first direct comparison of AAV-TT and AAV9 in the adult nonhuman primate brain and positions AAV-TT as a promising tool for disease modeling and therapeutic interventions in neurodegenerative disorders targeting anatomically distributed neural circuits.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"744-755"},"PeriodicalIF":4.6,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147377186","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":"Interview with Manlio Fusciello, PhD.","authors":"Thomas Gallagher","doi":"10.1177/10430342261471872","DOIUrl":"10.1177/10430342261471872","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"630-633"},"PeriodicalIF":4.6,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148470411","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":"The 2026 ESGCT Spring School.","authors":"Thomas Gallagher","doi":"10.1177/10430342261471871","DOIUrl":"https://doi.org/10.1177/10430342261471871","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":"37 15-16","pages":"639-640"},"PeriodicalIF":4.6,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148712195","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":"AAV Vectors Have Limited Impact on Host Chromatin Accessibility and Nuclear Architecture.","authors":"Joël W Zöllig, Maija K Pietilä, Melina O Bräuer, Kurt Tobler, Cornel Fraefel","doi":"10.1177/10430342261437975","DOIUrl":"10.1177/10430342261437975","url":null,"abstract":"<p><p>Recombinant adeno-associated virus (rAAV) vectors are widely used for gene therapeutics, yet their early effects on host chromatin remodeling and nuclear organization remain insufficiently characterized. In contrast, several DNA viruses are known to remodel host chromatin accessibility, for example, Herpes simplex virus type 1 (HSV-1) in mammalian cells and baculoviruses in insect systems. Here, we evaluated genome accessibility and nuclear organization of primary human fibroblast cells at 24 and 48 h after infection with wild-type AAV2, single-stranded (ss) and self-complementary rAAV2, as well as ssAAV2 and ssAAV-DJ in human lung carcinoma cells. Genome-wide assay for transposase-accessible chromatin using sequencing showed no detectable change in host chromatin accessibility at either time point. A DNase I digestion assay at five candidate loci supported this observation. Although host accessibility remained stable, immunofluorescence-based single-cell analyses uncovered modest changes in histone abundance, RNA polymerase II-associated signals, nuclear morphology, and the organization of liquid-liquid phase-separated nuclear compartments. These effects were generally mild in primary fibroblasts but showed greater dependence on vector type, cell cycle state, and transgene expression in carcinoma cells. Collectively, at the tested doses and times, AAV-based vectors did not remodel chromatin accessibility at scale and induced only small changes in polymerase-associated readouts and nuclear architecture. These data are consistent with limited nuclear perturbation under these conditions.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"756-769"},"PeriodicalIF":4.6,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147580221","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":"Dichotomies in <i>Ex Vivo</i> and <i>In Vivo</i> Performance of Receptor-Binding Mutants of Adeno-Associated Virus Vectors.","authors":"Olaniyi Olarewaju, Martin Bentler, Jacqueline Breuer, Fatih Alioglu, Anna Grabowski, Aileen Prosche, Haider Sami, Manfred Ogris, Hildegard Büning","doi":"10.1177/10430342261465461","DOIUrl":"10.1177/10430342261465461","url":null,"abstract":"<p><p>Receptor engagement plays a key role in the cellular uptake, intracellular trafficking, and overall transduction efficiency of adeno-associated virus (AAV) vectors. Although heparan sulfate proteoglycan (HSPG) and α5ß1 integrin-binding motifs of the AAV serotype 2 (AAV2) capsid were mapped, it has remained incompletely understood how loss of these interactions affects AAV vector performance. Hence, we generated capsid variants harboring mutations at capsid residues responsible for HSPG binding (AAV2ΔHSPG), α5ß1 integrin binding (AAV2ΔIntegrin), or both (AAV2ΔHSPGΔIntegrin), and investigated the mutants systematically <i>ex vivo</i> and <i>in vivo</i>. While neither production nor packaging efficiency was affected, variants revealed distinct physicochemical alterations, including altered electrophoretic mobility and thermal stability. <i>Ex vivo</i>, loss of HSPG binding completely abolished transgene expression across various cell lines, whereas ablation of α5ß1 integrin binding lowered transduction efficiency. While cellular uptake was reduced for AAV2ΔIntegrin and almost eliminated for AAV2ΔHSPG and AAV2ΔHSPGΔIntegrin, mutants nevertheless reached the nuclear compartment, albeit with lower efficiency compared with AAV2. Strikingly, <i>in vivo</i> performance diverged sharply from <i>ex vivo</i> findings as we observed a substantially enhanced transduction in multiple non-hepatic tissues for AAV2ΔHSPG and AAV2ΔHSPGΔIntegrin in C57BL/6N albino and BALB/c mice, as well as a strong liver detargeting, while AAV2ΔIntegrin was non-infectious <i>in vivo</i>. These data uncover a fundamental dichotomy between <i>ex vivo</i> and <i>in vivo</i> determinants of AAV2 transduction and identify receptor-binding ablation, especially integrin binding, as a potential alternative to detarget AAV2 vectors for next-generation capsid engineering and tissue-specific retargeting.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"788-801"},"PeriodicalIF":4.6,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148455662","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}