Human gene therapy最新文献

{"title":"FDA Issues Plausible Mechanism Pathway Draft Guidance.","authors":"Alex Philippidis","doi":"10.1177/10430342261437961","DOIUrl":"10.1177/10430342261437961","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"387-391"},"PeriodicalIF":4.0,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147580277","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":"Prime Editing of Alzheimer's Disease High-Risk APOE4 Allele by Brain-Directed Adeno-Associated Virus Vectors.","authors":"Caner Günaydin, Neil R Hackett, Victor Wakim, Dolan Sondhi, Stephen M Kaminsky, Ronald G Crystal","doi":"10.1177/10430342251401888","DOIUrl":"10.1177/10430342251401888","url":null,"abstract":"<p><p>Common variants of the apolipoprotein E (APOE) gene have a major impact on the risk of developing Alzheimer's disease (AD). Relative to homozygotes with the common E3 allele, the APOE4 variant (C112R) increases risk by 3.5-fold in E3/E4 heterozygotes and 15-fold in E4 homozygotes. Since the E3 and E4 alleles differ only by a single nucleotide, gene editing of E4 to E3 is a potential strategy to reduce AD risk in E4 homozygotes. Because the APOE pool in the brain is separate from systemic APOE, editing to treat AD would ideally be directed to the brain. Following <i>in vitro</i> optimization of prime editing guide RNAs, efficient prime editing expression cassettes were inserted into the adeno-associated virus (AAV) split-intein system and packaged into pairs of AAV vectors for <i>in vivo</i> editing. The AAV vectors were administered to human homozygous APOE4-targeted replacement mice (TRE4), and APOE4 to APOE3 editing efficiency was assessed after 4 weeks. The prime editing construct designated APOE3/4-3_10 was the most efficient at APOE4 to APOE3 conversion, both in liver following intravenous delivery and in brain following intrahippocampal delivery. To assess brain-wide editing, two AAV capsids were compared, including AAVrh.10 with administration either directly to the hippocampus or to the cerebrospinal fluid via the cisterna magna and AAV-CAP.B10 administered intravenously. Other than minor differences in APOE4/3-3_10 mediated E4 to E3 editing in the cerebellum, the different capsids and routes yielded similar editing efficacy throughout the brain. This may represent a candidate treatment to reduce the risk of AD.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"403-411"},"PeriodicalIF":4.0,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145833815","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":"<i>Corrigendum to:</i> Leveraging CRISPR-Cas9 for Accurate Detection of AAV-Neutralizing Antibodies: The AAV-HDR Method.","authors":"","doi":"10.1177/10430342261436594","DOIUrl":"https://doi.org/10.1177/10430342261436594","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"10430342261436594"},"PeriodicalIF":4.0,"publicationDate":"2026-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147638621","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":"Preferential Encapsulation of DNA Impurities Derived from pAAV Plasmid Backbone into Recombinant Adeno-Associated Virus Capsids.","authors":"Yuzhe Yuan, Kiyoko Higashiyama, Emi Ito-Kudo, Kyoko Masumi-Koizumi, Keisuke Yusa, Kazuhisa Uchida","doi":"10.1177/10430342251415385","DOIUrl":"10.1177/10430342251415385","url":null,"abstract":"<p><p>Recombinant adeno-associated viruses (rAAVs) are useful vectors for clinical gene therapy. It is crucial to examine DNA impurities, such as plasmid DNA, for quality control of rAAV products. In this study, we examined highly purified rAAV1, rAAV2, rAAV5, and rAAV6 samples produced on a three-plasmid platform, using a high-throughput sequencer. These samples contained 0.49-3.80% detectable DNA impurities derived from the three plasmids, as estimated by the ampicillin resistance gene (<i>amp</i>^R). The plasmid impurities consisted of 90.62-95.84% pAAV, 3.21-6.83% pRC, and 0.95-2.55% pHelper DNA. These trends were consistent with those of DNA impurities determined by droplet digital PCR (ddPCR), indicating that cleaved pAAV backbone DNA was the primary source of DNA impurities encapsulated into the capsids. To examine the preferential encapsulation of pAAV backbone DNA into capsids, short sequencing reads were mapped to the entire pAAV backbone sequence, and we found that the reads were relatively evenly distributed across the backbone sequence, with occasional sharp drops. Furthermore, the read length distribution containing the pAAV backbone sequence showed a main peak at 3.2 kb in Oxford Nanopore Technologies sequencing. This length was consistent with that of backbone DNA nicked at two terminal resolution sites by Rep78/68. Analysis of the terminal sequences of the long reads containing backbone sequences revealed that 92.7-98.4% of them contained Rep-binding elements. These results indicate that the pAAV backbone in linear DNA form was cleaved from pAAV-ZsGreen1 by Rep78/68 nicking at <i>trs</i> in the nucleus, and that the single-stranded DNA was efficiently translocated into the capsids through the encapsulating machinery, similar to the rAAV genome.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"349-364"},"PeriodicalIF":4.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146142213","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, Seamless Ink Up-to-$1.12B Hearing Loss Collaboration.","authors":"Alex Philippidis","doi":"10.1177/10430342261426584","DOIUrl":"10.1177/10430342261426584","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"295-299"},"PeriodicalIF":4.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147325297","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":"Comparative Analysis of Induced Pancreatic Stem Cells Generated with Different Factors.","authors":"Hirofumi Noguchi, Chika Miyagi-Shiohira, Takuya Sadahira, Masami Watanabe, Issei Saitoh","doi":"10.1177/10430342251414882","DOIUrl":"10.1177/10430342251414882","url":null,"abstract":"<p><p>Recently, our research group generated induced tissue-specific stem/progenitor (iTS/iTP) cells. Compared with induced pluripotent stem (iPS) cells, iTS/iTP cells offer several advantages, that is, easy generation, higher differentiation efficiency, and no teratoma formation. In this study, iTS cells were generated from mouse pancreatic tissues (iTS-P cells) using two different methods. Plasmid vectors were used for expressing <i>Oct3/4</i>, <i>Sox2</i>, <i>Klf4</i>, <i>c-Myc</i> (<i>OSKM</i>), or <i>Yap1</i> (<i>YAP</i>) to evaluate the efficiency and differentiation potential of the resulting cells. No significant difference in reprogramming efficiency between the <i>OSKM</i>- and <i>YAP</i>-based methods was observed. Among the established clones, iTS-P OSKM2 and iTS-P YAP9 cells, which demonstrated high efficiency in differentiating to insulin-producing cells (IPCs), were selected for further comparison. Both iTS-P OSKM2 and iTS-P YAP9 cells expressed genetic markers of endoderm and pancreatic progenitors and differentiated into IPCs more efficiently than the embryonic stem (ES) cells. Genomic bisulfite sequencing revealed that the pluripotency factors <i>Oct3/4</i> and <i>Nanog</i> were partially methylated in both iTS-P OSKM2 and iTS-P YAP9 cells. Unsupervised hierarchical clustering of gene expression profiles showed that iTS-P YAP9 cells clustered more closely with the ES cells than with the iTS-P OSKM2 cells. However, the expression levels of <i>Oct3/4</i> and <i>Nanog</i> were significantly lower in both iTS-P OSKM2 and YAP9 cells than in the ES cells. These results conclude that no substantial difference is present in the characteristics between iTS-P cells induced by <i>OSKM</i> or <i>YAP</i>, and that their higher differentiation efficiency than the ES cells indicates promising potential for clinical applications.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"329-340"},"PeriodicalIF":4.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146112853","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":"Less Invasive Light Irradiation Methods for Inner Ear Gene Therapy Based on Photoswitch Technology: Toward Clinical Translation.","authors":"Masao Noda, Takahiro Otabe, Ryota Koshu, Naomi Takino, Mika Ito, Makoto Ito, Fuun Kawano, Takahiro Nakajima, Moritoshi Sato, Shin-Ichi Muramatsu","doi":"10.1177/10430342251415191","DOIUrl":"10.1177/10430342251415191","url":null,"abstract":"<p><p>Hearing impairment, one of the most prevalent sensory disorders, remains a major risk factor for dementia in the aging population. Although interventions such as hearing aids and cochlear implants provide partial benefit, they do not address the underlying pathology of sensorineural hearing loss. Inner ear gene therapy has attracted significant attention as a promising approach; however, its clinical translation requires minimally invasive and controllable methods for gene activation. We previously developed a photoactivatable Cre recombinase (PA-Cre) system for spatiotemporal regulation of gene expression. In this study, we evaluated the feasibility of irradiating the external auditory canal (EAC) and tympanic membrane (TM) as minimally invasive approaches for activating cochlear gene expression. Tyrosine-mutant AAV9/3 vectors (AAV.GTX) encoding PA-Cre and a Cre-dependent reporter (sfGFP-to-tdTomato) were injected via the round window membrane in 9-week-old C57BL/6J mice. Seven days later, light irradiation was applied using three approaches: (1) Direct cochlear irradiation via postauricular access, (2) TM irradiation with a fiber-optic probe, and (3) noninvasive EAC irradiation through the intact TM. Recombination efficiency in inner hair cells (IHCs) was quantified using whole-mount immunohistochemistry. AAV.GTX efficiently transduced IHCs and drove robust sfGFP expression. In the absence of light, tdTomato expression remained minimal (<5%), indicating low basal Cre leak activity. Direct cochlear irradiation produced strong recombination (conversion rate: 88.4 ± 1.5%), confirming the functionality of PA-Cre in the mouse inner ear. TM and EAC irradiation yielded high conversion efficiencies (95.8 ± 1.7% and 97.6 ± 1.2%, respectively), comparable to direct irradiation, while preserving cochlear integrity. These findings indicate that PA-Cre functions effectively in the mouse cochlea with minimal leak activity and that TM and EAC irradiation enable robust, minimally invasive gene activation. This strategy highlights the light-mediated, noninvasive modulation of cochlear gene expression, informing future translational development.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"341-348"},"PeriodicalIF":4.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146112839","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":"Lipid Nanoparticle-Delivered mRNA Therapy Corrects Neonatal Murine MPS I-H.","authors":"Jiamei Fu, Mei Luo, Xiu Jin, Jing Su, Li Song, Qiuxia Xu, Fanfei Liu, Xiaoyi Wu, Min Luo, Qin Ye, Qiqi Li, Yifang An, Chengda Ren, Ming Hu, Man Liu, Manjun Li, Shaohua Yao, Yang Yang","doi":"10.1177/10430342251412392","DOIUrl":"10.1177/10430342251412392","url":null,"abstract":"<p><p>Mucopolysaccharidosis type I-Hurler (MPS I-H) is a severe lysosomal storage disorder caused by α-L-iduronidase (IDUA) deficiency, leading to glycosaminoglycan (GAG) accumulation and progressive multisystem dysfunction, including the central nervous system (CNS). Early hematopoietic stem cell transplantation remains the primary disease-modifying intervention for MPS I-H despite its clinical challenges. Conventional enzyme replacement therapy (ERT), which has limited CNS efficacy due to blood-brain barrier (BBB) restrictions, is being addressed by the development of novel BBB-penetrating ERT platforms. Recently, mRNA therapy has become a promising treatment option for rare genetic diseases by enabling the <i>in vivo</i> supplementation of defective or deficient proteins to ameliorate the disease phenotype. To evaluate mRNA therapy for MPS I-H, we screened ionizable lipids to develop lipid nanoparticle systems optimized for hepatic delivery, which were subsequently used to encapsulate mRNA encoding human IDUA (hIDUA) fused to a validated BBB-penetrating melanotransferrin peptide (MTfp). This formulation was intravenously administered to neonatal MPS I-H mice at a dose of 1.0 mg/kg every 2 weeks for 4 months. The treatment significantly increased IDUA enzyme activity in both the serum and liver. Concurrently, it reduced GAG accumulation in the urine, peripheral tissues, and partial CNS regions (olfactory bulbs, hippocampus, and cerebellum). Furthermore, the treatment significantly improved cardiac and skeletal development, as assessed by echocardiography and micro-computed tomography, respectively. Notably, it also enhanced cognitive function, as evidenced by the improved performance in the Delayed-Matching-to-Place dry maze test. Our findings demonstrate that LNP-MTfp-hIDUA treatment effectively ameliorates key pathological features and promotes neurodevelopment in MPS I-H, establishing a promising and safe therapeutic strategy for this disorder.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":"37 7-8","pages":"311-328"},"PeriodicalIF":4.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147511854","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":"An Analysis of Biomarkers for the Evaluation of Gene Therapy in Niemann-Pick Disease Type C1 Mice.","authors":"Chika Watanabe, Masamitsu Maekawa, Eriko Jimbo, Yoshie Kurokawa, Karin Kojima, Kazuhiro Muramatsu, Keitaro Miyoshi, Chen Wu, Miki Igarashi, Shin-Ichi Muramatsu, Hitoshi Osaka, Yoshikatsu Eto, Takanori Yamagata","doi":"10.1177/10430342251412391","DOIUrl":"10.1177/10430342251412391","url":null,"abstract":"<p><p>Niemann-Pick disease type C1 (NPC1) is an autosomal recessive lysosomal storage disorder caused by pathogenic variants of the <i>NPC1</i> gene that encodes a protein essential for lysosomal cholesterol transport. A deficiency in NPC1 results in the accumulation of unesterified cholesterol and sphingolipids, leading to neurological, psychiatric, and hepatic manifestations from infancy to adulthood. The currently approved treatment is palliative. Although the efficacy of gene therapy has been demonstrated in murine models, reliable biomarkers for evaluating the treatment effects remain unknown. We evaluated adeno-associated virus (AAV) vector-mediated NPC1 gene therapy in <i>Npc1</i> homo-knockout (<i>Npc1^-/-</i>) mice, focusing on blood-based biomarkers. An AAV vector carrying human <i>NPC1</i> under a cytomegalovirus promoter (AAV-<i>hNPC1</i>) was administered intraperitoneally on days 6-8 after birth at varying vector doses and analyzed at multiple time points: 1.8 × 10¹¹ vector genomes/mouse analyzed at 7 weeks (Low/7w) and 1.0 × 10¹² vector genomes/mouse at 4 weeks (High/4w) and 9 weeks (High/9w). <i>hNPC1</i> is expressed in the brain and liver, and a degree of neuronal cell survival is observed. High-dose AAV treatment improves body weight and rotarod performance. Plasma <i>N</i>-palmitoyl-<i>O</i>-phosphocholine-serine (PPCS) and lysosphingomyelin (lyso-SM) levels were significantly elevated in <i>Npc1^-/-</i> mice. PPCS increased with disease progression but was significantly decreased after later points of high-dose AAV treatment (saline-treated <i>Npc1^-/-</i> mice: 12.88 ± 3.53 ng/mL, AAV-treated <i>Npc1^-/-</i> mice: 7.87 ± 1.67 ng/mL, <i>p</i> = 0.0008). Lyso-SM and oxysterols showed limited changes after therapy. Vector genome analysis revealed higher and more sustained levels in the brain than in the liver, which is consistent with rapid hepatocyte proliferation-reducing vector persistence. These findings demonstrate that systemic AAV-<i>hNPC1</i> therapy ameliorates motor and neurological deficits but has a limited impact on several cholesterol-related biomarkers. PPCS has been suggested as a sensitive biomarker of therapeutic response and warrants further evaluations in preclinical and clinical NPC1 gene therapy trials.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":"37 7-8","pages":"300-310"},"PeriodicalIF":4.6,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147511811","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":"Exploring Development Options of a Polishing Chromatography Step for AAV7 and AAV8.","authors":"Xiaotong Fu, Colin Haws, Darren Begley, Richard Gyamfi Atta","doi":"10.1177/10430342251408498","DOIUrl":"10.1177/10430342251408498","url":null,"abstract":"<p><p>As impact of adeno-associated virus (AAV) empty capsids on drug product safety and quality remains inconclusive, downstream purification strategy has been focusing on empty capsid removal. Anion-exchange chromatography (AEX) has made significant progress in separating empty from full capsids in recent years. Still, achieving baseline resolution between different AAV subpopulations remains challenging due to subtle charge differences. With a certain AAV construct design, this difficulty is compounded when upstream packaging efficiency is low or when empty and full capsids of a particular serotype have similar electrostatic charge profiles. To improve separation and product purity, secondary interaction mechanisms using multimodal (mix-mode) chromatography are often introduced.In this study, we present a case study on developing a polishing chromatography step to remove empty capsids from AAV7 and AAV8 preparations. To create a challenging feed material for the polishing step, we used small gene-of-interest (GOI) and poorly packaged starting materials. We investigated multiple critical process parameters, including buffer matrix, salt concentration, pH, peak fractionation strategies, and column chemistry (strong AEX vs. mix-mode). Mass photometry (MP) and charge detection mass spectrometry (CDMS) were used to characterize capsid populations. Optimized AEX conditions for AAV8 achieved 80% full capsids by MP and 90% GOI-containing capsids by CDMS. For AAV7, the mix-mode column demonstrated improved resolution compared with the standard AEX gradient method. These results demonstrate that mix-mode chromatography provides an alternative polishing option for serotypes where traditional AEX fails to achieve the desired separation.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"372-382"},"PeriodicalIF":4.0,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145855742","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}