Human gene therapy最新文献

{"title":"Marks' Resignation Sparks Concerns on FDA Regulation of Gene Therapies.","authors":"Alex Philippidis","doi":"10.1089/hum.2025.075","DOIUrl":"10.1089/hum.2025.075","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"851-855"},"PeriodicalIF":3.9,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144012119","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":"A Single-Sector Higher Throughput Sedimentation Velocity Analytical Ultracentrifugation Method for Recombinant Adeno-Associated Virus Empty and Full Ratio Analysis.","authors":"Xiang Li, Qikun Yu, Hua Bi, Dening Pei, Da Zhang, Wei Jiang, Xiaodong Ye, Zhenzhen Cai, Wenxiu Hou, Akash Bhattacharya, Yichen Yang, Cong Wang, Miao Ye, Xi Qin, Dehua Huo, Chenggang Liang","doi":"10.1089/hum.2024.162","DOIUrl":"10.1089/hum.2024.162","url":null,"abstract":"<p><p>Recombinant adeno-associated virus (rAAV) has emerged as one of the most important gene delivery vectors in the field of gene therapy due to its unique advantages and characteristics. The empty and full ratio is a critical quality attribute in the quality control (QC) of rAAV, and its accurate evaluation is crucial for ensuring the safety, effectiveness, and consistency of gene therapy products. Analytical ultracentrifugation (AUC) technology, with its high resolution and accuracy, is widely recognized by the industry as the gold standard for identifying the empty and full ratio of rAAV. However, the conventional sedimentation velocity analytical ultracentrifugation (SV-AUC) method has limited throughput, failing to meet the large-scale detection needs of rAAV in process development and QC. This study aims to develop a single-sector higher throughput SV-AUC method without the need for a reference sector for blank control in order to improve the throughput of detecting the empty and full ratio of rAAV vectors. We optimized the traditional double-sector SV-AUC method, which requires a reference sector for blank control in the cell. By converting the light intensity data of AUC into pseudo-absorbance data, we significantly improve the analytical throughput. By tracking the variation of light intensity data with radius, we could clearly observe the sedimentation process of the rAAV sample. Despite a difference in the absolute value of pseudo-absorbance, the accurately fitted relative absorbance value and the traditional SV-AUC absorbance value with blank control were comparable, further verifying the applicability of this upgraded rAAV analytical method. The detailed comparison and verification between the upgraded method and the traditional SV-AUC method showed that the consistency and repeatability of the percentage and sedimentation coefficient were excellent both within the same cell and across different cells. The analysis results of samples from seven independent cells with a total of 14 sectors showed that the overall data exhibited good repeatability. The consistency of the high percentage empty capsid (HE) samples repeatability results was good, and the overlay of the C(s) distribution diagram also showed good pattern consistency. The relative standard deviation of the average percentage of empty, partial, and full capsids was maintained within 5%. The upgraded method demonstrated excellent consistency and repeatability in the analysis of rAAV samples with different empty and full ratios, aligning closely with the data obtained with the traditional SV-AUC method, the gold standard. Linear correlation analysis between the titers of HE samples and the overall absorbance (A value) of AUC, as well as the absorbance of empty, partial, and full capsids, revealed a good linear relationship, further confirming the applicability and reliability of the upgraded AUC method for evaluating rAAV samples with different titers. We also preliminarily e","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"925-936"},"PeriodicalIF":3.9,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144077363","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":"Halting Recombinant Adeno-Associated Virus Transgene Expression Using mRNA-Lipid Nanoparticle-Delivered Meganucleases.","authors":"Rubens Tavora, Lizhou Zhang, Mai H Tran, Hao Li, Dan O'Hagan, Andi Pan, Lorenzo Barrett, Joseph A Jablonski, Sonia Mediouni, Alexander Lopez, Zachary Comella, Charles Bailey, Hyeryun Choe, Michael Farzan, Susana T Valente","doi":"10.1089/hum.2025.011","DOIUrl":"10.1089/hum.2025.011","url":null,"abstract":"<p><p>Recombinant adeno-associated virus (rAAV) vectors are increasingly preferred for <i>in vivo</i> gene therapy due to their broad tropism, low immunogenicity, and sustained transgene expression. Nevertheless, in cases of adverse reactions to these expressions, a method to suppress or permanently halt rAAV transgene activity could significantly enhance the safety of these vectors. To address this need, we employed meganucleases-highly specific DNA endonucleases with long recognition sequences. By placing meganuclease target sites within rAAV transgenes, we created a system in which targeted cleavage leads to controlled disruption of transgene expression. Utilizing a luciferase assay, we screened various meganucleases and identified I-AniI-Y2, I-BmoI, and I-PpoI as prime candidates due to their high cleavage efficiencies. By strategically placing multiple meganuclease target sequences within introns, as well as in the 5' and 3' untranslated regions (UTRs) of transgenes, we significantly enhanced the cleavage efficiency of these meganucleases, ensuring robust and targeted suppression of transgene expression. Finally, we employed an mRNA-loaded lipid nanoparticledelivery system to demonstrate the ability of meganucleases to robustly inhibit rAAV-mediated transgene expression <i>in vitro</i>. Our findings underscore the potential of meganucleases as a viable safety mechanism in rAAV gene therapies, marking a significant advance toward safer long-term gene therapy approaches.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"870-883"},"PeriodicalIF":3.9,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12171709/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144011669","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":"Progress, Applications and Prospects of CRISPR-Based Genome Editing Technology in Gene Therapy for Cancer and Sickle Cell Disease.","authors":"Sha-Sha Zang, Ruirui Zhang, Jia-Run Zhang, Xi Zhang, Jun Li","doi":"10.1089/hum.2024.262","DOIUrl":"10.1089/hum.2024.262","url":null,"abstract":"<p><p>The advent of genome-editing technologies, particularly the RNA-guided the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated system (Cas) 9, which originates from prokaryotic CRISPR/Cas adaptive immune mechanisms, has revolutionized molecular biology. Renowned for its simplicity, cost-effectiveness, and capacity for multiplexed gene editing, CRISPR/Cas9 has emerged as the most versatile and widely adopted genome-editing platform. Its applications span fundamental research, biotechnology, medicine, and therapeutics. This review highlights recent advancements in CRISPR-based technologies, focusing on CRISPR/Cas9, CRISPR/Cas12a, and CRISPR/Cas12f. It emphasizes precision editing methods like base editing and prime editing, which enable targeted nucleotide changes without double-strand breaks. The specificity of these tools, including on-target accuracy and off-target risks, is critically evaluated. Additionally, recent preclinical and clinical efforts to treat diseases such as cancer and sickle cell disease using CRISPR are summarized. Finally, the challenges and future directions of CRISPR-mediated gene therapy are discussed, emphasizing its potential to integrate with other molecular approaches to address unmet medical needs.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"858-869"},"PeriodicalIF":3.9,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143965538","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":"BCMA-CAR Therapy for Multiple Myeloma in NOG Mice Prevents the Progression of Anemia and Bone Lesions.","authors":"Ryosuke Uchibori, Ken Ohmine, Takeshi Teruya, Junichi Mineno, Keiya Ozawa","doi":"10.1089/hum.2024.263","DOIUrl":"10.1089/hum.2024.263","url":null,"abstract":"<p><p>Multiple myeloma (MM) is an incurable hematological malignancy of plasma cells. Myeloma cells interfere with hematopoietic activities of the bone marrow, often leading to anemia, and can cause the bones to develop osteoporotic and lytic lesions. Clinical experience with chimeric antigen receptor T-cell (CAR-T) therapy targeting B-cell maturation antigen (BCMA) has been promising, with good response rates, favorable safety profiles, and low incidences of severe cytokine release syndrome and immune effector cell-associated neurotoxicity syndrome. However, CAR-T therapy in MM is accompanied by several new challenges, including therapeutic failure and relapse, and much attention has been paid to the further development of B-cell maturation antigen-chimeric antigen receptor (BCMA-CAR). Although most of the reported benefits of BCMA-CAR have been discussed, whether cancer can be eliminated, as well as the efficacy of CAR-T therapy for anemia and bone lesions, both myeloma-defining events, have not yet been reported in any animal model. In this study, we designed and verified a novel BCMA-specific chimeric antigen receptor (CAR). Our BCMA-CAR demonstrated the fundamental properties of CAR-T cells, including target-specific cytotoxic activity, cytokine production, and <i>in vivo</i> antitumor effects. In addition, we evaluated the therapeutic effect of BCMA-CAR in mice by imaging bone lesions and conducting blood examinations. Tumor mouse models showed systemic progression of MM in the bone marrow, and mice treated with saline or nongene modified T cells showed continued tumor progression, progressive bone lesions, and prolonged anemia. In contrast, all mice treated with gene modified T cells achieved a complete response, improved anemia to the level observed in normal mice, and suppressed progression of bone lesions. We concluded that anemia was improved with BCMA-CAR-T cell therapy. However, novel strategies to support the recovery of bone lesions by enhancing CAR-T cell function must be developed.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"902-913"},"PeriodicalIF":3.9,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144077480","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":"European Society of Gene & Cell Therapy Spring School 2025.","authors":"Thomas Gallagher","doi":"10.1089/hum.2025.088","DOIUrl":"10.1089/hum.2025.088","url":null,"abstract":"","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":"856-857"},"PeriodicalIF":3.9,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144101787","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":"Safe and Efficacious Permanent Removal of Large COL7A1 Exons for Gene Reframing as a Reliable Therapeutic Strategy for Recessive Dystrophic Epidermolysis Bullosa.","authors":"Sergio López-Manzaneda, Ángeles Mencía, José Bonafont, Alex Bassons-Bascuñana, Marta García, Alexander Nyström, Blanca Duarte, Sara Llames, Rodolfo Murillas, Silvia Modamio-Hoybjor, Matías Morín, Lucía Soletto, María J Escamez, Miguel A Moreno-Pelayo, Marcela Del Rio, Fernando Larcher","doi":"10.1089/hum.2024.238","DOIUrl":"https://doi.org/10.1089/hum.2024.238","url":null,"abstract":"<p><p>Mutations leading to premature termination codons in <i>COL7A1</i> are commonly associated with severe generalized recessive dystrophic epidermolysis bullosa (RDEB). Previous research, including our own, has indicated that removing mutated <i>COL7A1</i> exons along with the consequent reframing of <i>COL7A1</i> may not pose noticeable impact on protein function, offering a potential therapeutic strategy. However, investigations into the long-term <i>in vivo</i> effects of genome editing-mediated removal of mutant exons have only focused on the small exon 80 thus far. Hence, this study focuses on exons 73 and 105 of <i>COL7A1</i> to explore whether targeted exon removal, through a CRISPR/Cas9-assisted, Non-homologous end joining (NHEJ)-mediated approach, could be extended to other larger exons. Introducing ribonucleoprotein complexes carrying Cas9 and optimized sgRNA guide pairs for each exon (73 and 105) through electroporation efficiently led to their removal, consequently restoring type VII collagen (C7) synthesis in RDEB primary patient cells carrying frameshift mutations in these exons. <i>In vitro</i> tests indicated the normal stability of the resulting C7 variants expressed at physiological levels, while <i>in vivo</i> analyses of regenerated skin grafted onto immunodeficient mice using E73 or E105 RDEB edited cells demonstrated the proper deposition of C7 at the basement membrane zone, thereby restoring normal dermo-epidermal adherence. This study enhances the broader potential of the exon deletion approach in the treatment of RDEB.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144158326","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":"Stromal Gene Therapy Mediates Prolonged Protection Against Corneal Neovascularization Induced by an Aggressive Angiogenic Insult.","authors":"Mark Basche, Scott Robbie, D Frank P Larkin, Alexander J Smith, Rachael A Pearson, Robin R Ali","doi":"10.1089/hum.2024.248","DOIUrl":"https://doi.org/10.1089/hum.2024.248","url":null,"abstract":"<p><p>Corneal neovascularization (CoNV) is both a sight-threatening condition in and of itself and a major risk factor associated with corneal graft failure. Here, we determine the effectiveness of an adeno-associated viral vector (AAV)-based gene therapy targeting both hematic and lymphatic neovascularization in a murine model of severe CoNV. We first assessed the profile of transgene expression mediated by intrastromal injection of AAV2/8[Y733F] via longitudinal visualization of an enhanced Green Fluorescent Protein (eGFP) transgene and found that this serotype mediates a temporary (∼18 day) transduction of the corneal epithelium and sustained (≥148 day) transduction within the stroma. Constitutively expressed <i>sFlt1</i> or <i>sFlt4</i> were prophylactically delivered via intrastromal injection of AAV2/8[Y733F] vector at various intervals prior to aggressive induction of CoNV in a murine model. The extent of CoNV induced was quantified by fluorescein angiography and immunohistochemistry 17 days after induction. AAV2/8[Y733F]-CMV-sFlt1 was highly effective in the prevention of hemangiogenesis (HA) induced at 3, 28, and 210 days after intrastromal injection, but ineffective in the prevention of lymphangiogenesis. Two variants of AAV2/8[Y733F]-CMV-sFlt4 were ineffective in the prevention of angiogenesis when delivered alone, but combined delivery of AAV2/8[Y733F]-CMV-sFlt1 and AAV2/8[Y733F]-CMV-sFlt4 suggested a synergistic effect. Our results show that a single intrastromal injection of AAV2/8[Y733F]-CMV-sFlt1 is sufficient to protect against a robust stimulus for corneal HA over the long term. This technique could also be applied <i>ex vivo</i> to reduce the risk of failure in cases of \"high-risk\" corneal transplantation.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144158328","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":"A Highly Precise Method for the Quantitation of rAAV Cellular Uptake by ddPCR.","authors":"Albert Kiladjian, Prerana Pathak, Marina Feschenko, Svetlana Bergelson, Cullen Mason, Yu Wang","doi":"10.1089/hum.2025.023","DOIUrl":"https://doi.org/10.1089/hum.2025.023","url":null,"abstract":"<p><p>Recombinant adeno-associated virus (rAAV) has emerged as a leading vehicle for human gene therapy. An accurate and precise infectious titer assay is critical for assessing rAAV quality, potency, and product stability. The current gold standard for measuring rAAV infectivity is the median tissue culture infectivity dose (TCID50) method, which is laborious and highly variable. In the past several years, the droplet digital PCR (ddPCR) technology has made profound impacts on gene therapy analytics as it provides absolute DNA copy quantitation and is more accurate and precise than qPCR. In this article, we leveraged the ddPCR technology and developed a method to quantify rAAV cellular uptake <i>in vitro</i>. The results demonstrated that our method is consistent with TCID50 but is significantly more precise. Utilizing a stable AAV receptor (AAVR) cell line, this method can be implemented as a platform approach for various AAV serotypes and target genes. Moreover, the method is stability indicating, as desired for a potency assay. In conclusion, a novel rAAV uptake assay has been developed which reflects the mechanism of action of rAAV, and is accurate, precise and sensitive to product quality; thus overcoming many of the challenges of the traditional TCID50 method. It is particularly useful for initial rAAV product quality assessment and can contribute to a robust assay matrix with other product-specific potency assays for late-stage programs.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144158324","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":"Timely Intervention: Navigating Ethical Challenges in <i>OTOF</i>-Gene Therapy Trials.","authors":"Ellen Reisinger, Hans-Jörg Ehni, Oliver Feeney, Urban Wiesing","doi":"10.1089/hum.2025.042","DOIUrl":"https://doi.org/10.1089/hum.2025.042","url":null,"abstract":"<p><p><i>OTOF</i>-gene therapy for profound deafness in children has entered clinical trials. Given that there is an approved alternative therapy with cochlear implants, it is imperative to scrutinize the risks, while also highlighting the novel benefits, of this experimental gene therapy. Since the untreated inner ear subsequently degenerates in this form of inherited deafness, the <i>OTOF</i>-gene therapy will be most effective in young children. Moreover, the best outcome in terms of hearing and speech comprehension is expected when the gene therapy is applied before the age of 3 years. Given such \"earlier the better\" considerations, the optimal time for these clinical trials and this particular therapy is at an age when children are too young to give informed consent. Enrolling children, which are a vulnerable category of persons, in clinical trials where the balance of benefits and risks is uncertain, raises a series of ethical considerations. In this article, we outline how this research can be pursued in an ethically responsible manner.</p>","PeriodicalId":13007,"journal":{"name":"Human gene therapy","volume":" ","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144158331","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}