Human gene therapy最新文献

{"title":"The Application of Cell and Gene-Modified Cell Therapy in the Treatment of Osteopetrosis.","authors":"Jelena G Najdanović, Jelena M Živković, Stevo J Najman","doi":"10.1177/10430342261446500","DOIUrl":"10.1177/10430342261446500","url":null,"abstract":"<p><p>Bone tissue represents a dynamic tissue that undergoes continuous renewal and remodeling through the coupled actions of osteoblast-driven bone formation and osteoclast-mediated bone resorption. Proper coupling between these two processes is essential for maintaining bone homeostasis, whereas its disruption leads to skeletal pathology. Excessive osteoclast activity underlies disorders such as osteoporosis and Paget's disease, while osteoclast deficiency, either quantitative or functional, results in osteopetrosis. Osteopetrosis comprises a group of genetically heterogeneous and clinically variable rare metabolic bone diseases characterized by increased bone mass. Clinically, osteopetrosis is commonly classified into severe autosomal recessive osteopetrosis (ARO), also known as infantile malignant osteopetrosis, intermediate ARO, milder autosomal dominant osteopetrosis, and X-linked recessive osteopetrosis. Effective treatment options for both severe and milder forms of osteopetrosis remain an unresolved clinical challenge. This review sheds light on the potential of cell-based and gene-modified cell therapies as emerging strategies for the treatment of osteopetrosis. Current cell-based therapy approaches increasingly focus on induced pluripotent stem cells, while recent advances in gene therapy enable the correction of causative genetic defects through viral vector-mediated gene transfer, CRISPR/Cas9-based genome editing, or RNA interference-mediated gene silencing. In addition, the therapeutic potentials of immunological approaches based on recombinant human interferon gamma-1b is discussed. Despite significant progress, sustained research efforts are still required to translate cell and gene-modified cell therapy into effective and personalized clinical treatments for osteopetrosis.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"566-578"},"PeriodicalIF":4.0,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147928783","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":"Monitoring Fetal Somatic Cell Genome Editing <i>In Vivo</i> with Total-Body Positron Emission Tomography: Studies in Rhesus Macaques (<i>Macaca mulatta</i>).","authors":"Alice F Tarantal, Michele L Martinez, Lionel Sanz, Charles I Lee, He Yang, Henriette O'Geen, Dennis J Hartigan-O'Connor, David J Segal","doi":"10.1177/10430342261438941","DOIUrl":"10.1177/10430342261438941","url":null,"abstract":"<p><p>Translational development of somatic cell genome editing requires monitoring the extent of editing in the body at a given time, the specificity of editing, durability, and the potential for adverse events. A noninvasive approach that can identify edited cells <i>in vivo</i> is beneficial for addressing these and related questions. These studies used total-body positron emission tomography (PET) to identify somatic cell gene editing <i>in vivo</i> in fetal rhesus macaques. Rhesus dams were screened to confirm they were seronegative for AAV serotype 9 and <i>Sa</i>Cas9 antibodies, then selected for the study. Fetuses were administered a dual imaging vector (AAV9/<i>Sa</i>Cas9 and AAV9/<i>PCSK9</i> gRNA/HSV-sr39TK) <i>in utero</i> using an ultrasound-guided fetal intrahepatic approach in the second trimester (10¹² vector genomes/fetus). After maternal intravenous administration of 9-(4-(18)F-Fluoro-3-[hydroxymethyl]butyl)guanine ((18)F-FHBG) (∼3 mCi/kg), PET imaging was performed in the second and third trimesters. PET imaging provided evidence of editing in the fetal liver, which was sustained. Appropriate insertion of the promoterless HSV-sr39TK reporter in frame with the <i>PCSK9</i> gene was confirmed in the fetal liver near term using RNA sequencing, and correctly targeted insertions were observed. These studies have shown that total-body PET can provide insights into gene-edited somatic cells <i>in utero</i> and without evidence of adverse effects.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"579-587"},"PeriodicalIF":4.6,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13180422/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147689433","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":"Regeneron Wins FDA Approval for First Neurosensory Gene Therapy.","authors":"Alex Philippidis","doi":"10.1177/10430342261454500","DOIUrl":"10.1177/10430342261454500","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"547-550"},"PeriodicalIF":4.0,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148014994","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":"Harvest Process and Affinity Resin Selection Impacts on Adeno-Associated Virus Residual Host Cell Protein Retention.","authors":"Thomas M Leibiger, Lie Min, Kelvin H Lee","doi":"10.1177/10430342261443799","DOIUrl":"10.1177/10430342261443799","url":null,"abstract":"<p><p>Scalable purification platforms have been developed for adeno-associated virus (AAV) processing to support large-scale vector manufacturing. The ability of column chromatography to recover packaged vectors and remove empty capsids has been well-established, but knowledge gaps remain for understanding process parameter impacts on impurity retention. In this work, we examine the impacts of two key process parameters-the harvest method and affinity resin selection-on residual host cell protein (HCP) retention. Sequential window acquisition of all theoretical fragment ion mass spectra (SWATH-MS) proteomics is applied to comprehensively profile residual HCPs in affinity chromatography (AC) elution pools from four AAV serotypes (AAV2, -5, -8, and -9) produced by suspension HEK293 cells. Vectors were purified from cell culture lysates and from supernatants using POROS^™ CaptureSelect^™ AAVX (AAVX) and one additional serotype-specific affinity resin-Capto^™ AVB (AVB), POROS CaptureSelect AAV8 (PAAV8), or POROS CaptureSelect AAV9 (PAAV9). Significant divergence in residual HCP profiles was observed with the use of different affinity resins, with AVB and PAAV9 showing reduced residual HCP content in elution pools compared with AAVX and PAAV8. Processing of null culture lysates with fresh resins and resins with digested single-domain antibody fragments (sdAbs) shows that differences in resin performance are driven by variable nonspecific sdAb association with cellular impurities. Proteomic analysis of vector preparations from lysates compared with supernatants demonstrates product quality advantages of designing a media-only harvest process, specifically for AAV8, which was measured to contain an average of 66% of total vector genome content in the cell culture media. This work highlights the importance of serotype-specific tailoring of AAV downstream process design for improved product quality attributes to support clinical manufacture and small-scale analytics workflows.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"600-608"},"PeriodicalIF":4.0,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147698688","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-Mediated Base Editing for Correction of <i>RSPH4A</i> Mutations in Primary Ciliary Dyskinesia: A Proof-of-Concept Study.","authors":"Alessandro De Carli, Sara Pastore, Debora Maj, Fabio Filippini, Matteo Matteucci, Maria Elisa Di Cicco, Diego Peroni, Gabriele Donzelli, Stefania Crucitta, Lorena Zentilin, Angela Michelucci, Chiara Gabellini, Giulia Freer, Michele Lai, Massimo Pifferi, Mauro Pistello","doi":"10.1177/10430342261442511","DOIUrl":"10.1177/10430342261442511","url":null,"abstract":"<p><p>Primary ciliary dyskinesia (PCD) is a rare, genetically heterogeneous disorder, with abnormal ciliary motility, usually due to an ultrastructural defect, with chronic airway infections. Currently, no curative therapy exists for PCD. Given the prevalence of single nucleotide variants (SNVs) among causative mutations, we evaluated a novel base-editing approach. Specifically, we used a nickase Cas9 fused to adenosine deaminase to correct mutations in the radial spoke head component 4 A (<i>RSPH4A</i>) gene, causing PCD. We selected two PCD patients sharing the same SNV in <i>RSPH4A</i>, one with compound heterozygosity (child, patient 1) and one with homozygosity (adult, patient 2). After designing gRNAs, HEK293T cells with or without a DNA fragment containing the SNV in <i>RSPH4A</i>, were co-transfected with base editor plasmids. Complex formation and editing efficiency were validated by Western blot and digital PCR. We then treated patient cells with AAV containing the base editors and assessed ciliary beat frequency and motion pattern using high-speed video and confocal microscopy to evaluate delivery.Base editor complexes formed efficiently <i>in vitro</i>. AAV-mediated delivery in patient 1 cells led to an approximately 30.4% increase in normal motion pattern, with a corresponding reduction in circular motions (<i>p</i> < 0.001) compared with pre-treatment, and a 20% of editing efficiency detected by dPCR in transduced cells. Our data indicate that this limited editing efficiency is due to reduced AAV penetration in the lower layers of cells.This proof-of-concept study demonstrates the therapeutic potential of base editing for PCD, though current limitations include low editing efficiency and restricted delivery to inner cell layers in our experimental model. Future work should focus on optimizing base editors and testing novel delivery strategies to target progenitor cells, thereby enhancing the prospects for personalized gene therapy in PCD.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"588-599"},"PeriodicalIF":4.0,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147769862","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":"Strategies and Advances in Site-Specific Integration of Exogenous Large Genes.","authors":"Zhencheng Wu, Jia Chen, Manqi Huang, Wenqi Hu, Yiyu Liu, Hongwei Shao, Wenfeng Zhang","doi":"10.1177/10430342261445050","DOIUrl":"10.1177/10430342261445050","url":null,"abstract":"<p><p>Large genomic deletions (≥1 kb) are a recurrent class of disease-causing lesions in monogenic disorders, frequently leading to complete gene inactivation or the loss of critical cis-regulatory elements. Addressing these defects in a therapeutically relevant manner requires integration modalities capable of delivering and stably installing large exogenous DNA sequences at predefined genomic loci with an improved safety profile. By contrast, legacy approaches-including viral-vector delivery, recombinase-based strategies, and transposon-mediated insertion-typically achieve integration through random or semi-random mechanisms, which, despite their practicality and often favorable efficiencies, limit control over insertion site and copy number and may increase the risk of insertional mutagenesis and position-dependent variability in transgene expression. The past few years have witnessed rapid methodological diversification driven by genome editing, resulting in a growing repertoire of locus-specific strategies for large-fragment DNA insertion that are reshaping both disease-model construction and genetic therapeutics. In this Review, we synthesize the main classes of targeted large-fragment integration technologies reported to date. We begin with homology-directed repair (HDR)-dependent CRISPR-Cas9 knock-in strategies and discuss how donor architecture and local donor recruitment can be leveraged to improve integration outcomes for kilobase-scale payloads. We then examine approaches centered on prime editing, particularly those that couple prime editing with engineered serine/tyrosine recombinases to support programmable insertion of large DNA cargos. We close by surveying emerging HDR-independent systems based on CRISPR-guided transposition and retrotransposition, and we provide a comparative perspective on their performance envelopes, constraints, and trajectories toward broader biomedical applications.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"551-565"},"PeriodicalIF":4.0,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147769900","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":"Optimization of Alpha-1 Antitrypsin Expression from Adeno-Associated Virus Vectors.","authors":"Qiushi Tang, Debora Pires-Ferreira, Christian Mueller, Alisha M Gruntman, Terence R Flotte","doi":"10.1177/10430342261444637","DOIUrl":"10.1177/10430342261444637","url":null,"abstract":"<p><p>Alpha-1 antitrypsin deficiency (AATD) is an inherited disorder caused by mutations in <i>SERPINA1</i> that result in insufficient circulating alpha-1 antitrypsin (AAT) and progressive lung and liver diseases. Adeno-associated virus (AAV)-mediated gene therapy offers the potential for durable AAT expression; however, achieving therapeutic serum concentrations (≥11 µM) at clinically acceptable vector doses remains a major challenge. Here, we evaluated multiple AAV vector design strategies to enhance AAT expression and increase vector potency, thereby reducing the required dose to levels below those associated with severe adverse events. Using AAV1- and AAV8-based platforms, we compared promoter and enhancer configurations, codon optimization of the <i>SERPINA1</i> transgene, single-stranded versus self-complementary vector genomes, alternative polyadenylation signals, and an engineered oxidation-resistant AAT variant. Across mouse and ferret models, the chicken β-actin expression cassette consistently produced higher AAT levels than a liver-specific promoter variant despite comparable vector biodistribution, reflecting superior intrinsic transcriptional activity or an important contribution. Codon optimization did not enhance expression and, in some cases, modestly reduced AAT levels. Self-complementary AAV vectors exhibited reduced overall expression due to required promoter truncation, yielding lower transgene output than full-length single-stranded constructs. Modifications to polyadenylation signals or enhancer combinations did not improve expression. An oxidation-resistant AAT variant resulted in lower circulating levels but may retain therapeutic potential through enhanced functional stability. Collectively, these findings demonstrate that promoter strength and cassette architecture are dominant determinants of AAV-AAT potency and that full-length, single-stranded vectors with robust regulatory elements provide the highest expression. This work defines key parameters governing AAT expression <i>in vivo</i> and provides a foundation for next-generation AAV designs aimed at achieving therapeutic efficacy at safer, lower vector doses for the treatment of AATD.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"609-617"},"PeriodicalIF":4.0,"publicationDate":"2026-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147972013","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> \"Enhancing AAV9-UFμDys1 Gene Therapy Efficacy Through Immunosuppression in Mice with Pre-Existing Immunity and Enabling Redosing Strategies for Duchenne Muscular Dystrophy\".","authors":"","doi":"10.1177/10430342261456423","DOIUrl":"https://doi.org/10.1177/10430342261456423","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"10430342261456423"},"PeriodicalIF":4.0,"publicationDate":"2026-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148251304","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":"AVLAYAH and KRESLADI Win FDA Accelerated Approvals.","authors":"Alex Philippidis","doi":"10.1177/10430342261445261","DOIUrl":"10.1177/10430342261445261","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"459-462"},"PeriodicalIF":4.0,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147728903","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":"Adeno-Associated Virus Gene Delivery to Hard-to-Transduce Tissues: Biological Barriers, Engineering Strategies, and Clinical Challenges.","authors":"Xiaoyu Nan, Jiawen Sun, Yunuo Jiao, Wangan Li, Yujuan Chen, Yurong Liu, Bo Xu, Li Yu, Wenyan Guo, Yang Wu","doi":"10.1177/10430342261432669","DOIUrl":"10.1177/10430342261432669","url":null,"abstract":"<p><p>Adeno-associated virus (AAV) is widely regarded as a leading vector for gene therapy, underscored by clinical successes such as Luxturna and Zolgensma. However, efficient gene delivery to hard-to-transduce tissues-including the retina, deep skeletal muscle, and the central nervous system-remains a significant challenge, limited by structural barriers, preexisting immunity, and dose-dependent toxicities. This review systematically outlines recent advances in overcoming these delivery bottlenecks. We delve into four key strategic areas: (i) capsid engineering (<i>e.g.</i>, rational design, directed evolution, and computational approaches) to enhance tropism and evade immune detection; (ii) innovative delivery routes (local, systemic, and physical/chemical methods) to improve vector bioavailability; (iii) modulation of intracellular trafficking to boost nuclear delivery; and (iv) immunomodulatory strategies to mitigate both innate and adaptive immune responses. We further highlight translational progress in neuromuscular and retinal diseases and discuss persistent challenges. Looking forward, we envision that the convergence of next-generation capsids, smart vector systems, and integrated delivery platforms will be critical to expand the therapeutic landscape of AAVs from rare monogenic disorders to broader clinical applications.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"463-481"},"PeriodicalIF":4.6,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147615747","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}