Molecular Therapy-Methods & Clinical Development最新文献_第8页

Embryo and fetal gene editing: Technical challenges and progress toward clinical applications 胚胎和胎儿基因编辑：技术挑战与临床应用进展

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-03-04 DOI: 10.1016/j.omtm.2024.101229

Citra N.Z. Mattar, Wei Leong Chew, Poh San Lai

{"title":"Embryo and fetal gene editing: Technical challenges and progress toward clinical applications","authors":"Citra N.Z. Mattar, Wei Leong Chew, Poh San Lai","doi":"10.1016/j.omtm.2024.101229","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101229","url":null,"abstract":"Gene modification therapies (GMTs) are slowly but steadily making progress toward clinical application. As the majority of rare diseases have an identified genetic cause, and as rare diseases collectively affect 5% of the global population, it is increasingly important to devise gene correction strategies to address the root causes of the most devastating of these diseases and to provide access to these novel therapies to the most affected populations. The main barriers to providing greater access to GMTs continue to be the prohibitive cost of developing these novel drugs at clinically relevant doses, subtherapeutic effects, and toxicity related to the specific agents or high doses required. strategy and treating younger patients at an earlier course of their disease could lower these barriers. Although currently regarded as niche specialties, prenatal and preconception GMTs offer a robust solution to some of these barriers. Indeed, treating either the fetus or embryo benefits from economy of scale, targeting pre-pathological tissues in the fetus prior to full pathogenesis, or increasing the likelihood of complete tissue targeting by correcting pluripotent embryonic cells. Here, we review advances in embryo and fetal GMTs and discuss requirements for clinical application.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140156561","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Peptide-encoding gene transfer to modulate intracellular protein‒protein interactions 肽编码基因转移调节细胞内蛋白质与蛋白质之间的相互作用

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-02-28 DOI: 10.1016/j.omtm.2024.101226

Toshihiko Taya, Daisuke Kami, Fumiya Teruyama, Satoaki Matoba, Satoshi gojo

{"title":"Peptide-encoding gene transfer to modulate intracellular protein‒protein interactions","authors":"Toshihiko Taya, Daisuke Kami, Fumiya Teruyama, Satoaki Matoba, Satoshi gojo","doi":"10.1016/j.omtm.2024.101226","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101226","url":null,"abstract":"Peptide drug discovery has great potential, but the cell membrane is a major obstacle when the target is an intracellular protein‒protein interaction (PPI). It is difficult to target PPIs with small molecules; indeed, there are no intervention tools that can target any intracellular PPI. In this study, we developed a platform that enables the introduction of peptides into cells via mRNA-based gene delivery. Peptide-length nucleic acids do not enable stable ribosome binding and exhibit little to no translation into protein. In this study, a construct was created in which the sequence encoding dihydrofolate reductase (DHFR) was placed in front of the sequence encoding the target peptide, together with a translation skipping sequence, as a sequence that meets the requirements of promoting ribosome binding and rapid decay of the translated protein. This enabled efficient translation from the mRNA encoding the target protein while preventing unnecessary protein residues. Using this construct, we showed that it can inhibit Drp1/Fis1 binding, one of the intracellular PPIs, which governs mitochondrial fission, an important aspect of mitochondrial dynamics. In addition, it was shown to inhibit pathological hyperfission, normalize mitochondrial dynamics and metabolism, and inhibit apoptosis of the mitochondrial pathway.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140037415","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Manufacturing DNA in E. coli yields higher fidelity DNA than in vitro enzymatic synthesis 与体外酶法合成相比，在大肠杆菌中制造 DNA 可获得保真度更高的 DNA

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-02-28 DOI: 10.1016/j.omtm.2024.101227

Steven J. Hersch, Siddarth Chandrasekaran, Jamie Lam, Nafiseh Nafissi, Roderick A. Slavcev

{"title":"Manufacturing DNA in E. coli yields higher fidelity DNA than in vitro enzymatic synthesis","authors":"Steven J. Hersch, Siddarth Chandrasekaran, Jamie Lam, Nafiseh Nafissi, Roderick A. Slavcev","doi":"10.1016/j.omtm.2024.101227","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101227","url":null,"abstract":"Biotechnologies such as gene therapy have brought DNA vectors to the forefront of pharmaceuticals. The quality of starting material plays a pivotal role in determining final product quality. Here we examined the fidelity of DNA replication using enzymatic methods () compared to plasmid DNA produced in . Next-generation sequencing approaches rely on polymerases, which have inherent limitations in sensitivity. To address this challenge, we introduce a novel assay based on loss-of-function (LOF) mutations in the conditionally toxic gene. Our findings show that DNA production in results in significantly fewer LOF mutations (80- to 3000-fold less) compared to enzymatic DNA replication methods such as PCR and rolling circle amplification (RCA). These results suggest that using DNA produced by PCR or RCA may introduce a substantial number of mutation impurities, potentially affecting the quality and yield of final pharmaceutical products. Our study underscores that DNA synthesized has a significantly higher mutation rate than DNA produced traditionally in . Therefore, utilizing enzymatically-produced DNA in biotechnology and biomanufacturing may entail considerable fidelity-related risks, while using DNA starting material derived from substantially mitigates this risk.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140036798","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Endovascular transplantation of mRNA-enhanced mesenchymal stromal cells results in superior therapeutic protein expression in swine heart 通过血管内移植 mRNA 增强间充质基质细胞，猪心脏的治疗性蛋白表达效果更佳

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-02-27 DOI: 10.1016/j.omtm.2024.101225

Jonathan Al-Saadi, Mathias Waldén, Mikael Sandell, Jesper Solmér, Rikard Grankvist, Ida Friberger, Agneta Andersson, Mattias Carlsten, Kenneth Chien, Johan Lundberg, Nevin Witman, Staffan Holmin

{"title":"Endovascular transplantation of mRNA-enhanced mesenchymal stromal cells results in superior therapeutic protein expression in swine heart","authors":"Jonathan Al-Saadi, Mathias Waldén, Mikael Sandell, Jesper Solmér, Rikard Grankvist, Ida Friberger, Agneta Andersson, Mattias Carlsten, Kenneth Chien, Johan Lundberg, Nevin Witman, Staffan Holmin","doi":"10.1016/j.omtm.2024.101225","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101225","url":null,"abstract":"Heart failure has a poor prognosis and no curative treatment exists. Clinical trials are investigating gene- and cell-based therapies to improve cardiac function. The safe and efficient delivery of these therapies to solid organs is challenging. Herein, we demonstrate the feasibility of using an endovascular intramyocardial delivery approach to safely administer mRNA drug products and perform cell transplantation procedures in swine. Using a -vessel wall (TW) device, we delivered chemically modified mRNAs (modRNA) and mRNA-enhanced mesenchymal stromal cells expressing vascular endothelial growth factor A (VEGF-A) directly to the heart. We monitored and mapped the cellular distribution, protein expression, and safety tolerability of such an approach. The delivery of modRNA-enhanced cells via the TW device with different flow rates and cell concentrations marginally affect cell viability and protein expression . Implanted cells were found within the myocardium for at least 3 days following administration, without the use of immunomodulation and minimal impact on tissue integrity. Finally, we could increase the protein expression of VEGF-A over 500-fold in the heart using a cell-mediated modRNA delivery system compared with modRNA delivered in saline solution. Ultimately, this method paves the way for future research to pioneer new treatments for cardiac disease.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140097485","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Automated manufacture of ΔNPM1 TCR-engineered T cells for AML therapy 自动制造用于急性髓细胞性白血病治疗的 ΔNPM1 TCR 工程 T 细胞

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-02-27 DOI: 10.1016/j.omtm.2024.101224

Isabella Elias Yonezawa Ogusuku, Vera Herbel, Simon Lennartz, Caroline Brandes, Eva Argiro, Caroline Fabian, Carola Hauck, Conny Hoogstraten, Sabrina Veld, Lois Hageman, Karin Teppert, Georgia Koutsoumpli, Marieke Griffioen, Nadine Mockel-Tenbrinck, Thomas Schaser, Rosa de Groot, Ian C.D. Johnston, Dominik Lock

{"title":"Automated manufacture of ΔNPM1 TCR-engineered T cells for AML therapy","authors":"Isabella Elias Yonezawa Ogusuku, Vera Herbel, Simon Lennartz, Caroline Brandes, Eva Argiro, Caroline Fabian, Carola Hauck, Conny Hoogstraten, Sabrina Veld, Lois Hageman, Karin Teppert, Georgia Koutsoumpli, Marieke Griffioen, Nadine Mockel-Tenbrinck, Thomas Schaser, Rosa de Groot, Ian C.D. Johnston, Dominik Lock","doi":"10.1016/j.omtm.2024.101224","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101224","url":null,"abstract":"Acute myeloid leukemia (AML) is a heterogeneous malignancy that requires further therapeutic improvement, especially for the elderly and for subgroups with poor prognosis. A recently discovered T cell receptor (TCR) targeting mutant nucleophosmin 1 (ΔNPM1) presents an attractive option for the development of a cancer antigen-targeted cellular therapy. Manufacturing of TCR-modified T cells, however, is still limited by a complex, time-consuming, and laborious procedure. Therefore, this study specifically addressed the requirements for a scaled manufacture of ΔNPM1-specific T cells in an automated, closed, and good manufacturing practice-compliant process. Starting from cryopreserved leukapheresis, 2E8 CD8-positive T cells were enriched, activated, lentivirally transduced, expanded, and finally formulated. By adjusting and optimizing culture conditions, we additionally reduced the manufacturing time from 12 to 8 days while still achieving a clinically relevant yield of up to 5.5E9 ΔNPM1 TCR-engineered T cells. The cellular product mainly consisted of highly viable CD8-positive T cells with an early memory phenotype. ΔNPM1-TCR CD8 T cells manufactured with the optimized process showed specific killing of AML and . The process has been implemented in an upcoming phase 1/2 clinical trial for the treatment of NPM1-mutated AML.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140097488","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

B Cell Focused Transient Immune Suppression Protocol for Efficient AAV Readministration to the Liver 聚焦 B 细胞的瞬时免疫抑制方案，实现高效的 AAV 肝脏再管理

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-02-20 DOI: 10.1016/j.omtm.2024.101216

Jyoti Rana, Roland W. Herzog, Maite Muñoz-Melero, Kentaro Yamada, Sandeep RP. Kumar, Ahn K. Lam, David M. Markusic, Dongsheng Duan, Cox Terhorst, Barry J. Byrne, Manuela Corti, Moanaro Biswas

引用次数: 0

Preexisting antibody assays for gene therapy: Considerations on clinical cut-off for patient selection and companion diagnostic requirements 用于基因治疗的现有抗体检测：考虑患者选择的临床临界值和辅助诊断要求

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-02-20 DOI: 10.1016/j.omtm.2024.101217

Manuela Braun, Claudia Lange, Philipp Schatz, Brian Long, Johannes Stanta, Boris Gorovits, Edit Tarcsa, Vibha Jawa, Tong-Yuan Yang, Wibke Lembke, Nicole Miller, Fraser McBlane, Louis Christodoulou, Daisy Yuill, Mark Milton

引用次数: 0

Glutaredoxin-1 modulates the NF-κB signaling pathway to activate inducible nitric oxide synthase in experimental necrotizing enterocolitis 谷胱甘肽-1调节NF-κB信号通路，激活实验性坏死性小肠结肠炎中的诱导型一氧化氮合酶

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-02-19 DOI: 10.1016/j.omtm.2024.101214

Yunfei Zhang, Mei Yan, Yingying Xia, Yingbin Yue, Shuli Wang, Yuhui Hu, Genjian Lai, Quanjiang Wu, Qianyang Liu, Xin Ding, Chunbao Guo

引用次数: 0

Thorough molecular configuration analysis of non-canonical AAV genomes in AAV vector preparations 对 AAV 载体制备中的非经典 AAV 基因组进行全面的分子构型分析

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-02-19 DOI: 10.1016/j.omtm.2024.101215

Junping Zhang, Xiangping Yu, Matthew Chrzanowski, Jiahe Tian, Derek Pouchnik, Ping Guo, Roland W. Herzog, Weidong Xiao

引用次数: 0

A toxicology study of Csf2ra complementation and pulmonary macrophage transplantation therapy of hereditary PAP in mice Csf2ra 互补和肺巨噬细胞移植治疗遗传性 PAP 小鼠毒理学研究

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-02-17 DOI: 10.1016/j.omtm.2024.101213

Paritha Arumugam, Brenna C. Carey, Kathryn A. Wikenheiser-Brokamp, Jeffrey Krischer, Matthew Wessendarp, Kenjiro Shima, Claudia Chalk, Jennifer Stock, Yan Ma, Diane Black, Michelle Imbrogno, Margaret Collins, Dan Justin Kalenda Yombo, Haripriya Sakthivel, Takuji Suzuki, Carolyn Lutzko, Jose A. Cancelas, Michelle Adams, Elizabeth Hoskins, Dawn Lowe-Daniels, Lilith Reeves, Anne Kaiser, Bruce C. Trapnell

{"title":"A toxicology study of Csf2ra complementation and pulmonary macrophage transplantation therapy of hereditary PAP in mice","authors":"Paritha Arumugam, Brenna C. Carey, Kathryn A. Wikenheiser-Brokamp, Jeffrey Krischer, Matthew Wessendarp, Kenjiro Shima, Claudia Chalk, Jennifer Stock, Yan Ma, Diane Black, Michelle Imbrogno, Margaret Collins, Dan Justin Kalenda Yombo, Haripriya Sakthivel, Takuji Suzuki, Carolyn Lutzko, Jose A. Cancelas, Michelle Adams, Elizabeth Hoskins, Dawn Lowe-Daniels, Lilith Reeves, Anne Kaiser, Bruce C. Trapnell","doi":"10.1016/j.omtm.2024.101213","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101213","url":null,"abstract":"Pulmonary macrophage transplantation (PMT) is a gene and cell transplantation approach in development as therapy for hereditary pulmonary alveolar proteinosis (hPAP), a surfactant accumulation disorder caused by mutations in (and murine homologs). We conducted a toxicology study of PMT of gene-corrected macrophages (mGM-RαMϕs) or saline-control intervention in or wild-type (WT) mice including single ascending dose and repeat ascending dose studies evaluating safety, tolerability, pharmacokinetics, and pharmacodynamics. Lentiviral-mediated cDNA transfer restored GM-CSF signaling in mGM-RαMϕs. Following PMT, mGM-RαMϕs engrafted, remained within the lungs, and did not undergo uncontrolled proliferation or result in bronchospasm, pulmonary function abnormalities, pulmonary or systemic inflammation, anti-transgene product antibodies, or pulmonary fibrosis. Aggressive male fighting caused a similarly low rate of serious adverse events in saline- and PMT-treated mice. Transient, minor pulmonary neutrophilia and exacerbation of pre-existing hPAP-related lymphocytosis were observed 14 days after PMT of the safety margin dose but not the target dose (5,000,000 or 500,000 mGM-RαMϕs, respectively) and only in mice but not in WT mice. PMT reduced lung disease severity in mice. Results indicate PMT of mGM-RαMϕs was safe, well tolerated, and therapeutically efficacious in mice, and established a no adverse effect level and 10-fold safety margin.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":null,"pages":null},"PeriodicalIF":4.7,"publicationDate":"2024-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140569488","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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