Molecular Therapy最新文献

Viral and Cellular Insulators Promote Sustained HSV Vector Mediated Transgene Expression in Brain.

IF 12.1 1区医学

Molecular Therapy Pub Date : 2025-02-27 DOI: 10.1016/j.ymthe.2025.02.039

Selene Ingusci, Justus B Cohen, Joseph C Glorioso

{"title":"Viral and Cellular Insulators Promote Sustained HSV Vector Mediated Transgene Expression in Brain.","authors":"Selene Ingusci, Justus B Cohen, Joseph C Glorioso","doi":"10.1016/j.ymthe.2025.02.039","DOIUrl":"https://doi.org/10.1016/j.ymthe.2025.02.039","url":null,"abstract":"We have developed a gene therapy platform based on non-toxic, high-capacity replication defective (rd) herpes simplex virus type 1 (HSV-1) vectors. We previously determined that transgene expression from rdHSV-1 vectors requires strategic placement of insulators-small DNA elements that overcome the host's epigenetic silencing of foreign DNA-to maintain transgenes in euchromatin regions. Transgene expression was rescued by replacing either the latency associated transcript (LAT) or the the infected cell protein 4 (ICP4) gene with the transgene cassette close to naturally occurring viral insulators. The ICP4 locus was more permissive for transgene expression than the LAT locus in neurons in vitro. Following in vivo brain delivery, transgene expression from both loci lasted for at least 4 months. However, the level of expression tended to decline over time. To enhance transgene expression, we designed a novel insulator environment by combining cellular insulators with the resident viral insulators. In combination, these elements provided significantly higher levels of transgene expression in the brain than the viral insulators alone, lasting for at least 11.7 months. This new cassette design extends transgene activity in neurons compared to previous designs and holds promise for gene therapy applications in treating brain disorders.","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143531681","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A Universal Viral Capsid Protein Based One Step RNA Synthesis and Packaging System for Rapid and Efficient mRNA Vaccine Development.

IF 12.1 1区医学

Molecular Therapy Pub Date : 2025-02-27 DOI: 10.1016/j.ymthe.2025.02.037

Jiayue Su, Jinsong Zhang, Xiangning Feng, Jinsong Liu, Shan Gao, Xinrui Liu, Mingwei Yang, Zeliang Chen

{"title":"A Universal Viral Capsid Protein Based One Step RNA Synthesis and Packaging System for Rapid and Efficient mRNA Vaccine Development.","authors":"Jiayue Su, Jinsong Zhang, Xiangning Feng, Jinsong Liu, Shan Gao, Xinrui Liu, Mingwei Yang, Zeliang Chen","doi":"10.1016/j.ymthe.2025.02.037","DOIUrl":"https://doi.org/10.1016/j.ymthe.2025.02.037","url":null,"abstract":"The success of COVID-19 mRNA vaccines highlights the transformative potential of mRNA technology. Current mRNA vaccine development involves complex steps, including plasmid construction, RNA transcription, 5' capping, poly(A) tailing, and lipid nanoparticle encapsulation, yet challenges in vaccine accessibility persist. Here, we present an innovative mRNA platform leveraging the self-assembly capabilities of the MS2 bacteriophage viral capsid protein (VCP). A dual-promoter plasmid has been designed where one promoter drives VCP expression while the other transcribes target RNA containing pac sites, enabling rapid mRNA self-assembly in Escherichia coli (E. coli). Using an ovalbumin (OVA)-based tumor model, we validate the efficacy of this system. Tumor growth is significantly inhibited, accompanied by robust immune activation. Flow cytometry analyses reveal increased frequencies of OVA-specific CD8+, as well as activated and memory T cells. Additionally, the MS2-OVA vaccine favorably modulated the tumor immunosuppressive microenvironment by reducing myeloid-derived suppressor cells, while sustained antibody responses demonstrated the platform's ability to induce durable humoral immunity. These findings establish the feasibility of one-step mRNA synthesis and packaging in E. coli, providing a versatile and rapid platform for mRNA vaccine development, with broad implications for addressing global vaccination challenges.","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143531701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A differentiated β-globin gene replacement strategy uses heterologous introns to restore physiological expression.

IF 12.1 1区医学

Molecular Therapy Pub Date : 2025-02-27 DOI: 10.1016/j.ymthe.2025.02.036

Kirby A Wallace, Trevor L Gerstenberg, Craig L Ennis, Juan A Perez-Bermejo, James R Partridge, Christopher Bandoro, William M Matern, Gaia Andreoletti, Kristina Krassovsky, Shaheen Kabir, Cassandra D Lalisan, Aishwarya R Churi, Glen M Chew, Lana Corbo, Jon E Vincelette, Timothy D Klasson, Brian J Silva, Yuri G Strukov, B Joy Quejarro, Kaisle A Hill, Sebastian Treusch, Jane L Grogan, Daniel P Dever, Matthew H Porteus, Beeke Wienert

{"title":"A differentiated β-globin gene replacement strategy uses heterologous introns to restore physiological expression.","authors":"Kirby A Wallace, Trevor L Gerstenberg, Craig L Ennis, Juan A Perez-Bermejo, James R Partridge, Christopher Bandoro, William M Matern, Gaia Andreoletti, Kristina Krassovsky, Shaheen Kabir, Cassandra D Lalisan, Aishwarya R Churi, Glen M Chew, Lana Corbo, Jon E Vincelette, Timothy D Klasson, Brian J Silva, Yuri G Strukov, B Joy Quejarro, Kaisle A Hill, Sebastian Treusch, Jane L Grogan, Daniel P Dever, Matthew H Porteus, Beeke Wienert","doi":"10.1016/j.ymthe.2025.02.036","DOIUrl":"https://doi.org/10.1016/j.ymthe.2025.02.036","url":null,"abstract":"β-hemoglobinopathies are common monogenic disorders. In sickle cell disease (SCD) a single mutation in the β-globin (HBB) gene results in dysfunctional hemoglobin protein, while in β-thalassemia, over 300 mutations distributed across the gene reduce β-globin levels and cause severe anemia. Genetic engineering replacing the whole HBB gene through homology directed repair (HDR) is an ideal strategy to restore a benign genotype and rescue HBB expression for most genotypes. However, this is technically challenging because 1) the insert must not be homologous to the endogenous gene and 2) synonymous codon-optimized, intron-less sequences may not reconstitute adequate β-globin levels. Here, we developed an HBB gene replacement strategy using CRISPR-Cas9 that successfully addresses these challenges. We determined that a DNA donor containing a diverged HBB coding sequence and heterologous introns to avoid sequence homology provides proper physiological expression. We identified a DNA donor that uses truncated γ-globin introns, results in 34% HDR, rescues β-globin expression in in vitro models of SCD and β-thalassemia in hematopoietic stem and progenitor cells (HSPCs). Furthermore, while HDR allele frequency dropped in vivo, it was maintained at ∼15%, demonstrating editing of long-term repopulating HSPCs. In summary, our HBB gene replacement strategy offers a differentiated approach by restoring naturally regulated adult hemoglobin expression.","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143531697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

CAR-mediated target recognition limits TCR-mediated target recognition of TCR- and CAR-dual-receptor-edited T cells.

IF 12.1 1区医学

Molecular Therapy Pub Date : 2025-02-27 DOI: 10.1016/j.ymthe.2025.02.035

Tassilo L A Wachsmann, Teuntje Poortvliet, Miranda H Meeuwsen, Dennis F G Remst, Marijke F Toes, Anne K Wouters, Renate S Hagedoorn, J H Frederik Falkenburg, Mirjam H M Heemskerk

{"title":"CAR-mediated target recognition limits TCR-mediated target recognition of TCR- and CAR-dual-receptor-edited T cells.","authors":"Tassilo L A Wachsmann, Teuntje Poortvliet, Miranda H Meeuwsen, Dennis F G Remst, Marijke F Toes, Anne K Wouters, Renate S Hagedoorn, J H Frederik Falkenburg, Mirjam H M Heemskerk","doi":"10.1016/j.ymthe.2025.02.035","DOIUrl":"https://doi.org/10.1016/j.ymthe.2025.02.035","url":null,"abstract":"Antigen escape can compromise the efficacy of chimeric antigen receptor (CAR-) or T cell receptor (TCR-) engineered T cells. Targeting multiple antigens can effectively limit antigen escape, and combining CAR- with TCR-mediated targeting can significantly broaden the spectrum of targetable antigens. Here, we explored whether dual-antigen specificity can be installed on T cells using combined TCR- and CAR-engineering to prevent antigen escape of multiple myeloma (MM). We report the generation of CD8 T cells that were transduced to express a transgenic TCR, targeting a peptide derived from transcriptional coactivator BOB1 in the context of HLA-B*07:02, alongside a BCMA-targeting CAR. Those T cells, termed TRaCR T cells, efficiently recognized target cells that were resistant to either BOB1 TCR or BCMA CAR T cells, illustrating general dual-specificity. In the presence of both antigens however, target cell recognition was preferentially conferred via the CAR, compromising TCR-mediated target cell recognition. Importantly, this resulted in a survival advantage for tumor cells lacking expression of BCMA in an in vivo model of heterogenous MM. In conclusion, we demonstrate general dual-specificity of TRaCR T cells but advise caution when using TRaCR T cells as strategy to target heterogenous tumors.","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143531623","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Gene Therapy Then and Now: A Look Back at Changes in the Field Over the Past 25 Years.

IF 12.1 1区医学

Molecular Therapy Pub Date : 2025-02-27 DOI: 10.1016/j.ymthe.2025.02.040

Dan Wang, Gregg Stevens, Terence R Flotte

引用次数: 0

A VSV-based oral rabies vaccine was sentineled by Peyer's patches and induced a timely and durable immune response.

IF 12.1 1区医学

Molecular Therapy Pub Date : 2025-02-27 DOI: 10.1016/j.ymthe.2025.02.038

Shen Wang, Zhenshan Wang, Weiqi Wang, Hongyu Sun, Na Feng, Yongkun Zhao, Jianzhong Wang, Tiecheng Wang, Xianzhu Xia, Feihu Yan

{"title":"A VSV-based oral rabies vaccine was sentineled by Peyer's patches and induced a timely and durable immune response.","authors":"Shen Wang, Zhenshan Wang, Weiqi Wang, Hongyu Sun, Na Feng, Yongkun Zhao, Jianzhong Wang, Tiecheng Wang, Xianzhu Xia, Feihu Yan","doi":"10.1016/j.ymthe.2025.02.038","DOIUrl":"https://doi.org/10.1016/j.ymthe.2025.02.038","url":null,"abstract":"The global eradication of canine-mediated human rabies remains an ongoing public health priority. While conventional oral rabies vaccines (ORVs) have demonstrated partial success in interrupting zoonotic transmission, current formulations necessitate improvement in both immunogenic profiles and mechanistic clarity. Herein, we present a recombinant vesicular stomatitis virus (VSV)-vectored vaccine candidate (rVSVΔG-ERA-G) engineered to express the glycoprotein of the rabies virus (RABV) ERA strain, substituting the native VSV glycoprotein. Preclinical evaluation across multiple mammalian species (Mus musculus, Canis lupus familiaris, Felis catus, Arctic fox, and Nyctereutes procyonoides) revealed rapid seroconversion and sustained neutralizing antibody responses. Challenge experiments demonstrated 100% survival efficacy in pre-exposure prophylaxis models, with partial protection observed in post-exposure scenarios. Safety assessments confirmed significant attenuation of neurotropism and absence of horizontal transmission or environmental shedding. Furthermore, evidence shown that rVSVΔG-ERA-G is recognized by Peyer's patches (PPs), where a cascade activation of immune cells occurred. From another perspective, the absence of functional microfold cells in PPs hampered the initiation and progression of immune responses. This proof-of-concept study establishes rVSVΔG-ERA-G as an ORV candidate with enhanced biosafety and cross-species immunogenicity. The elucidation of M cell-dependent mucosal priming mechanisms provides a rational framework for optimizing the targeted delivery of ORVs.","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143531617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Genetically reprogrammed exosomes for cancer immunotherapy.

IF 12.1 1区医学

Molecular Therapy Pub Date : 2025-02-25 DOI: 10.1016/j.ymthe.2025.02.009

Wenxing Gu, Qingchang Tian, Tian Xie, Xiaoyuan Chen

引用次数: 0

The impact of revised regenerative medicine regulations on unapproved gene therapies in Japan.

IF 12.1 1区医学

Molecular Therapy Pub Date : 2025-02-25 DOI: 10.1016/j.ymthe.2025.02.018

Yuichiro Ukon, Satoshi Hosoya, Kazuki Morita, Eisuke Minegishi, Jun Sugihara

引用次数: 0

Addressing barriers to clinical translation of extracellular vesicle therapeutics.

IF 12.1 1区医学

Molecular Therapy Pub Date : 2025-02-25 DOI: 10.1016/j.ymthe.2025.02.020

Steven M Jay

引用次数: 0

GMP manufacturing of cell and gene therapy products: Challenges, opportunities, and pathways forward.

IF 12.1 1区医学

Molecular Therapy Pub Date : 2025-02-25 DOI: 10.1016/j.ymthe.2025.02.022

Brian Fury, Gerhard Bauer

{"title":"GMP manufacturing of cell and gene therapy products: Challenges, opportunities, and pathways forward.","authors":"Brian Fury, Gerhard Bauer","doi":"10.1016/j.ymthe.2025.02.022","DOIUrl":"https://doi.org/10.1016/j.ymthe.2025.02.022","url":null,"abstract":"Cell and gene therapy (CGT) products have been emerging as life-saving and life-changing therapies over the last 20 years. The United States has been a forerider in the development of these therapeutic products and has also developed the pertinent manufacturing methods for these complicated \"living medicines.\" California has emerged as a prominent hub for CGT manufacturing, hosting a variety of good manufacturing practice (GMP) facilities. These facilities are pivotal in advancing CGT products from phase 1 to phase 3 clinical trials and, eventually, to commercialization. Despite California's strategic advantages, including its biotech ecosystem and access to venture capital, numerous challenges hinder its full potential. These include funding disparities, expertise limitations, stringent regulatory demands, and a mismatch between facility utilization and demand. This review explores these factors more in depth and provides a comprehensive analysis of the current state of GMP manufacturing for CGTs in California. The example of the current situation in the state of California may also serve as an analogy for other states; we chose California due to our decades of experience manufacturing CGT products in this state.","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":" ","pages":""},"PeriodicalIF":12.1,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143516170","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0