Molecular Therapy-Methods & Clinical Development最新文献

To be or not to PHP.eB? Potential strategies for therapeutically "attacking" multiple sclerosis. 是还是不是PHP.eB？治疗“攻击”多发性硬化症的潜在策略。

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2025-09-01 eCollection Date: 2025-09-11 DOI: 10.1016/j.omtm.2025.101574

Jessica A Herstine, Benjamin L L Clayton

引用次数: 0

Erratum: Preclinical evaluation of the efficacy and safety of AAV1-hOTOF in mice and nonhuman primates. 勘误：AAV1-hOTOF在小鼠和非人灵长类动物中的有效性和安全性的临床前评估。

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2025-08-22 eCollection Date: 2025-09-11 DOI: 10.1016/j.omtm.2025.101575

Longlong Zhang, Hui Wang, Mengzhao Xun, Honghai Tang, Jinghan Wang, Jun Lv, Biyun Zhu, Yuxin Chen, Daqi Wang, Shaowei Hu, Ziwen Gao, Jianping Liu, Zheng-Yi Chen, Bing Chen, Huawei Li, Yilai Shu

引用次数: 0

Effects of CRISPR-Cas9-mediated FOXP3 knockout on CAR T cell potency. crispr - cas9介导的FOXP3敲除对CAR - T细胞效力的影响

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2025-08-21 eCollection Date: 2025-09-11 DOI: 10.1016/j.omtm.2025.101570

Lena Peter, Martí Farrera-Sal, Ferhat Ali Yaman, Nils Henrik Dempewolf, Samira Picht, Sarah Schulenberg, Jonas Kath, Frederik Hamm, Frederik Heinrich, Dimitrios L Wagner, Mir-Farzin Mashreghi, Annette Künkele, Petra Reinke, Julia K Polánsky, Michael Schmueck-Henneresse

{"title":"Effects of CRISPR-Cas9-mediated FOXP3 knockout on CAR T cell potency.","authors":"Lena Peter, Martí Farrera-Sal, Ferhat Ali Yaman, Nils Henrik Dempewolf, Samira Picht, Sarah Schulenberg, Jonas Kath, Frederik Hamm, Frederik Heinrich, Dimitrios L Wagner, Mir-Farzin Mashreghi, Annette Künkele, Petra Reinke, Julia K Polánsky, Michael Schmueck-Henneresse","doi":"10.1016/j.omtm.2025.101570","DOIUrl":"10.1016/j.omtm.2025.101570","url":null,"abstract":"Persistent antigen stimulation and inflammatory environments drive exhaustion, senescence, and activation-induced cell death, impairing both endogenous and therapeutic T cells. Understanding the mechanisms underlying T cell dysfunction is critical for improving immunotherapies. While the transcription factor forkhead box protein P3 (FOXP3) is primarily known for its role in regulatory T cell development and maintenance, recent studies suggest it may also influence effector T cell function. However, its impact on therapeutic T cells, including CAR T cells, remains poorly defined. Here, we used non-viral CRISPR-Cas9 editing to knockout FOXP3 in CD19-directed CAR T cell products (TCPs) generated via lentiviral transduction. FOXP3 expression was upregulated at both the protein and RNA level following CAR stimulation. Compared to unmodified CAR TCPs, FOXP3-KO CAR TCPs showed comparable exhaustion profiles but enhanced cytokine production and prolonged cytotoxic function across repeated antigen challenges. These findings identify FOXP3 as a context-dependent modulator of CAR T cell function and suggest that its disruption may enhance therapeutic potency without exacerbating exhaustion. FOXP3 targeting may represent a complementary strategy to improve the functional resilience of CAR T cell therapies in cancer or autoimmune disease.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"33 3","pages":"101570"},"PeriodicalIF":4.7,"publicationDate":"2025-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12433514/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145070730","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Simple, specific, rapid, and pharmacopoeia-compliant qPCR approach for the detection of mycoplasma in biopharmaceuticals. 简便、特异、快速、符合药典要求的qPCR方法用于生物制药中支原体的检测。

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2025-08-20 eCollection Date: 2025-09-11 DOI: 10.1016/j.omtm.2025.101572

Sergio Dos Santos, Emilie Lespinasse, Baptiste Bonnet, Stéphane Basmaciogullari

{"title":"Simple, specific, rapid, and pharmacopoeia-compliant qPCR approach for the detection of mycoplasma in biopharmaceuticals.","authors":"Sergio Dos Santos, Emilie Lespinasse, Baptiste Bonnet, Stéphane Basmaciogullari","doi":"10.1016/j.omtm.2025.101572","DOIUrl":"10.1016/j.omtm.2025.101572","url":null,"abstract":"Cell substrate utilized in the production of biologics intended for human use need to be cleared from mycoplasma contaminant as described in pharmacopoeia. While the gold-standard method for mycoplasma detection involves lengthy microbiology and cell-based assays, nucleic acid technologies offer the possibility to accelerate testing. In this study, we develop a qPCR assay capable of specifically detecting 11 mycoplasma species relevant to pharmacopoeias with only two primers and two hydrolysis probes, which greatly facilitates operations. While the primers cross-react with bacterial species, the specificity conferred by the hydrolysis probes allows for confident interpretation of the qPCR result and unambiguous statement regarding the presence of mycoplasma in the test article. Amplicon sequencing can further confirm the identity of contaminants. This comprehensive assay can therefore be of great help to quality control laboratories embedded in biologics production sites, which need to provide results in a timely manner and contribute to root cause analysis in case of contamination.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"33 3","pages":"101572"},"PeriodicalIF":4.7,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12441703/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145088112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Durable tissue-specific transgene expression in newborn mice following intraperitoneal delivery of non-cytotoxic HSV vectors. 腹腔内递送无细胞毒性HSV载体后新生小鼠的持久组织特异性转基因表达。

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2025-08-20 eCollection Date: 2025-09-11 DOI: 10.1016/j.omtm.2025.101573

Yoshitaka Miyagawa, Motoyo Maruyama, Atsushi Sakai, Yuriko Sato, Masumi Shimizu, Seiji Kuroda, Hiromi Hayashita-Kinoh, Motoko Yamamoto, Ryotaro Hashizume, Hidenori Suzuki, Justus B Cohen, Joseph C Glorioso, Takashi Okada

{"title":"Durable tissue-specific transgene expression in newborn mice following intraperitoneal delivery of non-cytotoxic HSV vectors.","authors":"Yoshitaka Miyagawa, Motoyo Maruyama, Atsushi Sakai, Yuriko Sato, Masumi Shimizu, Seiji Kuroda, Hiromi Hayashita-Kinoh, Motoko Yamamoto, Ryotaro Hashizume, Hidenori Suzuki, Justus B Cohen, Joseph C Glorioso, Takashi Okada","doi":"10.1016/j.omtm.2025.101573","DOIUrl":"10.1016/j.omtm.2025.101573","url":null,"abstract":"This report describes the distribution and transgene expression of two non-cytotoxic, replication-defective (rd) herpes simplex virus (HSV) vectors, JΔNI7 and JΔNI8, following intraperitoneal delivery to newborn mice. The two vectors are functionally defective for all immediate-early genes, and JΔNI8 is further deleted for the UL41 endonuclease (vhs). Both vectors were engineered to express a red luciferase gene from the LAT locus to track vector distribution and gene expression in vivo. A comparison of reporter gene activities under the control of four different promoters in JΔNI7 showed that the strongest expression was achieved with the CAG promoter. Distribution analysis at 1 week post-injection showed transgene expression in multiple tissues, but at 4 weeks, high-level expression was limited to the spinal cord, skin, and muscles. JΔNI8 showed rapid clearance of vector DNA in most tissues, suggesting a role for the vhs gene in vector stability. Compared with wild-type KOS strain injections, JΔNI7-based non-cytotoxic rdHSV did not induce substantial CD45+ immune-cell infiltration or tissue destruction, suggesting that our rdHSV vectors are safe. Taken together, these results demonstrate tissue-specific, durable transgene expression following systemic delivery of rdHSV vectors, suggesting their potential for systemic gene therapy for newborns with skin or neuromuscular diseases.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"33 3","pages":"101573"},"PeriodicalIF":4.7,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12446203/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145115127","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Safety through design: Expanding options for spinal muscular atrophy gene therapy. 安全性通过设计：扩大脊髓性肌萎缩症基因治疗的选择。

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2025-08-19 eCollection Date: 2025-09-11 DOI: 10.1016/j.omtm.2025.101550

Ewout J N Groen, Renske I Wadman

引用次数: 0

Natural history of preexisting AAV5 antibodies in adults with hemophilia B during the lead-in of the etranacogene dezaparvovec phase 3 study. 在etranacogene dezaparvovec 3期研究开始期间，成人血友病B患者中预先存在的AAV5抗体的自然历史

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2025-08-18 eCollection Date: 2025-09-11 DOI: 10.1016/j.omtm.2025.101568

Robert Klamroth, Michael Recht, Nigel S Key, Wolfgang Miesbach, Steven W Pipe, Radoslaw Kaczmarek, Douglass Drelich, Blanca Salazar, Sandra Le Quellec, Paul E Monahan, Nicholas Galante, Paul van der Valk, Jacqueline Tarrant

{"title":"Natural history of preexisting AAV5 antibodies in adults with hemophilia B during the lead-in of the etranacogene dezaparvovec phase 3 study.","authors":"Robert Klamroth, Michael Recht, Nigel S Key, Wolfgang Miesbach, Steven W Pipe, Radoslaw Kaczmarek, Douglass Drelich, Blanca Salazar, Sandra Le Quellec, Paul E Monahan, Nicholas Galante, Paul van der Valk, Jacqueline Tarrant","doi":"10.1016/j.omtm.2025.101568","DOIUrl":"10.1016/j.omtm.2025.101568","url":null,"abstract":"Testing for binding or neutralizing antibodies (NAbs) to adeno-associated virus (AAV) is part of the laboratory assessment of people with hemophilia considering AAV-based gene therapy. We evaluated the natural history of NAb titers to AAV serotype 5 (AAV5) in adult males ≥18 years old with hemophilia B (factor IX ≤ 2%) during the lead-in period of a phase 3 trial prior to the infusion of etranacogene dezaparvovec to characterize NAb in addition to immunoglobulin G (IgG) and immunoglobulin M (IgM) anti-AAV5 binding antibody changes over time. At screening, 48% (32/67) of enrolled participants had detectable NAbs (NAb+) with a median titer of 58 (range: 9-3,440). Participant-specific lead-in periods differed and included discontinuers (median duration: 240 days; range: 1-360). The median intra-participant coefficient of variation of NAb titer over time was 25% (range: 2%-154%). NAb seropositivity was associated with older age (p = 0.0065). For participants with detectable anti-AAV5 NAbs and IgG, there was a high correlation of titers at each visit (median r = 0.96; range: 0.92-0.99). IgM anti-AAV5 antibodies were detectable in only 9% of participants, and seroconversion was infrequent. In conclusion, AAV5 NAb test results were consistent over 6 months, which informs the timing of NAb screening when considering gene therapy for hemophilia B.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"33 3","pages":"101568"},"PeriodicalIF":4.7,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12441693/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145088146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

High-potency MyoAAV capsids enhanced skeletal muscle correction in a mouse model of GSD IIIa. 高效MyoAAV衣壳在GSD IIIa小鼠模型中增强骨骼肌矫正。

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2025-08-18 eCollection Date: 2025-09-11 DOI: 10.1016/j.omtm.2025.101567

Kuo-An Liao, Sang-Oh Han, Mercedes Barzi, Haiqing Yi, William Eisner, Beatrice Bissig-Choisat, Karl-Dimiter Bissig, Baodong Sun

{"title":"High-potency MyoAAV capsids enhanced skeletal muscle correction in a mouse model of GSD IIIa.","authors":"Kuo-An Liao, Sang-Oh Han, Mercedes Barzi, Haiqing Yi, William Eisner, Beatrice Bissig-Choisat, Karl-Dimiter Bissig, Baodong Sun","doi":"10.1016/j.omtm.2025.101567","DOIUrl":"10.1016/j.omtm.2025.101567","url":null,"abstract":"Glycogen storage disease type IIIa (GSD IIIa) affects multiple tissues, including liver, heart, and skeletal muscles. We recently reported that an adeno-associated virus serotype 9 vector expressing pullulanase, a bacterial glycogen debranching enzyme, driven by an immunotolerizing dual promoter (AAV9-Dual-Pull), effectively decreased pullulanase-induced cytotoxic T lymphocyte response and corrected disease abnormalities in all major affected tissues in GSD IIIa mice. To reduce effective vector dosages for transgene delivery to skeletal muscles, we packaged the AAV-Dual-Pull vector into two muscle-tropic MyoAAV capsids, MyoAAV4A and MyoAAV4E. Six weeks after administration of the same dose vector (1 × 1013 vg/kg), both the MyoAAV vectors demonstrated remarkably greater transduction efficiency and glycogen clearance efficacy in the cardiac and skeletal muscles than the AAV9 vector, accompanied by the improvement of muscle function, reversal of liver abnormalities, and normalization of the disease biomarker, Glc4 in the urine. Furthermore, treatment with the MyoAAV4A-Dual-Pull vector at a 10-fold lower dose (1×1012 vg/kg) achieved significantly better therapeutic outcomes in the skeletal muscles than the AAV9-Dual-Pull vector at a high dose (1×1013 vg/kg). Validation in human liver chimeric mice revealed that the MyoAAV vectors and the AAV9 vectors had a similar efficiency in transducing human hepatocytes, indicating increased translatability for clinical applications.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"33 3","pages":"101567"},"PeriodicalIF":4.7,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12419012/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145042308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Harnessing mRNA-lipid nanoparticles as innovative therapies for autoimmune diseases. 利用mrna -脂质纳米颗粒作为自身免疫性疾病的创新疗法。

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2025-08-18 eCollection Date: 2025-09-11 DOI: 10.1016/j.omtm.2025.101566

Rose Razavi, Michael Kegel, Jenna Muscat-Rivera, Drew Weissman, Jilian R Melamed

{"title":"Harnessing mRNA-lipid nanoparticles as innovative therapies for autoimmune diseases.","authors":"Rose Razavi, Michael Kegel, Jenna Muscat-Rivera, Drew Weissman, Jilian R Melamed","doi":"10.1016/j.omtm.2025.101566","DOIUrl":"10.1016/j.omtm.2025.101566","url":null,"abstract":"Autoimmune diseases are chronic conditions where the immune system mistakenly attacks healthy tissues, leading to potentially debilitating symptoms that require lifelong management. There are no cures for autoimmune diseases, and new treatments are urgently needed to improve patient outcomes. This review delves into the compelling advancements and ongoing challenges in harnessing mRNA-lipid nanoparticles (LNPs) as innovative therapies for autoimmune diseases. mRNA-LNPs enable a range of therapeutic approaches to combat autoimmune diseases, including targeted immune cell modulation, tissue regeneration, antigen-specific tolerizing immunotherapy, and in vivo chimeric antigen T cell therapies. To successfully advance this promising class of therapies to the clinic, key challenges must be addressed, such as mitigating unwanted inflammation caused by LNPs, overcoming biological barriers to delivery, and ensuring the long-term safety of mRNA-LNPs specifically in autoimmune contexts. Through their modular design, flexible application, and potential for cost-effective production, mRNA-LNP therapies offer exciting clinical potential to transform the management of autoimmune diseases.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"33 3","pages":"101566"},"PeriodicalIF":4.7,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12441705/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145088098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Nanoparticle delivery of AMPK activator 991 prevents its toxicity and improves muscle homeostasis in Duchenne muscular dystrophy. 纳米颗粒递送AMPK激活剂991可防止其毒性并改善杜氏肌营养不良患者的肌肉稳态。

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2025-08-14 eCollection Date: 2025-09-11 DOI: 10.1016/j.omtm.2025.101564

Ilaria Andreana, Ananga Ghosh, Mathieu Repellin, Anita Kneppers, Sabrina Ben Larbi, Federica Tifni, Aurélie Fessard, Marion Martin, Jacqueline Sidi-Boumedine, David Kryza, Barbara Stella, Silvia Arpicco, Claire Bordes, Yves Chevalier, Julien Gondin, Bénédicte Chazaud, Rémi Mounier, Giovanna Lollo, Gaëtan Juban

{"title":"Nanoparticle delivery of AMPK activator 991 prevents its toxicity and improves muscle homeostasis in Duchenne muscular dystrophy.","authors":"Ilaria Andreana, Ananga Ghosh, Mathieu Repellin, Anita Kneppers, Sabrina Ben Larbi, Federica Tifni, Aurélie Fessard, Marion Martin, Jacqueline Sidi-Boumedine, David Kryza, Barbara Stella, Silvia Arpicco, Claire Bordes, Yves Chevalier, Julien Gondin, Bénédicte Chazaud, Rémi Mounier, Giovanna Lollo, Gaëtan Juban","doi":"10.1016/j.omtm.2025.101564","DOIUrl":"10.1016/j.omtm.2025.101564","url":null,"abstract":"Muscular dystrophies, such as Duchenne muscular dystrophy (DMD), are caused by permanent muscle injuries leading to chronic inflammation, with macrophages harboring an altered inflammatory profile contributing to fibrosis through the secretion of transforming growth factor β1 (TGF-β1). We previously showed that AMP-activated protein kinase (AMPK) activation reduces TGF-β1 secretion by macrophages and improves muscle homeostasis and muscle force in a DMD mouse model. However, direct AMPK activators like compound 991 show strong adverse effects in vivo. To overcome this toxicity, we encapsulated 991 into biodegradable polymeric poly(lactic-co-glycolic) acid (PLGA) nanoparticles for in vivo delivery. We show that 991-loaded PLGA nanoparticles retained drug activity on fibrotic macrophages in vitro and in vivo. In the D2-mdx DMD mouse model, intravenously injected PLGA nanoparticles reached macrophages in gastrocnemius and diaphragm muscles, two severely affected muscles in this model, but not in heart and quadriceps. Chronic intravenous injections of 991-loaded PLGA nanoparticles decreased inflammation in both gastrocnemius and diaphragm, which was associated with TGF-β1 level and fibrosis reduction and increase in myofiber size and muscle mass in the gastrocnemius, without toxicity. These results demonstrate that nanomedicine is an efficient strategy to deliver AMPK activators in vivo to target inflammation and improve the dystrophic muscle phenotype in the gastrocnemius.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"33 3","pages":"101564"},"PeriodicalIF":4.7,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12433517/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145070773","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0