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

Engineering single-cycle MeV vector for CRISPR-Cas9 gene editing 为 CRISPR-Cas9 基因编辑设计单循环 MeV 载体

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-06-24 DOI: 10.1016/j.omtm.2024.101290

Ramya Rallabandi, Brenna Sharp, Spencer Majerus, Austin Royster, Sarrianna Hoffer, Mia Ikeda, Patricia Devaux

引用次数: 0

Highly branched poly β-amino ester/CpG-depleted CFTR plasmid nanoparticles for non-viral gene therapy in lung cystic fibrosis disease 用于肺囊性纤维化疾病非病毒基因疗法的高支化聚 β-氨基酯/CpG-depleted CFTR 质粒纳米颗粒

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-06-24 DOI: 10.1016/j.omtm.2024.101292

Bei Qiu, Darío Manzanares, Yinghao Li, Xianqing Wang, Zishan Li, Sébastien Terreau, Zhonglei He, Jing Lyu, Wenxin Wang, Irene Lara-Sáez

{"title":"Highly branched poly β-amino ester/CpG-depleted CFTR plasmid nanoparticles for non-viral gene therapy in lung cystic fibrosis disease","authors":"Bei Qiu, Darío Manzanares, Yinghao Li, Xianqing Wang, Zishan Li, Sébastien Terreau, Zhonglei He, Jing Lyu, Wenxin Wang, Irene Lara-Sáez","doi":"10.1016/j.omtm.2024.101292","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101292","url":null,"abstract":"Lung cystic fibrosis (CF) is a lethal inherited disease caused by mutations in the CF transmembrane conductance regulator () gene, leading to a dysfunctional CFTR protein. Gene therapy offers promise for the treatment of lung CF. However, the development and clinical application of CF gene therapy have long been hampered by the absence of safe and highly efficient delivery vectors. In this work, a novel polymer-based gene replacement treatment approach was developed. A series of poly (β-amino esters) (PAEs) with various topological structures and chemical compositions were screened to create non-viral therapeutic systems for CFTR restoration in lung CF disease. A nanoparticle, formed by the selected highly branched PAE (HPAE) with a CpG-depleted CFTR plasmid, demonstrated gene expression and biocompatibility in lung epithelial cells, outperforming leading commercial gene transfection reagents such as Lipofectamine 3000 and Xfect. The newly developed gene therapy system successfully restored functional CFTR protein production in lung CF epithelial monolayers. This therapeutic approach holds great potential for use as an efficient and safe non-viral treatment for CF patients.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"16 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141552024","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

Lipid nanoparticle-encapsulated DOCK11-siRNA efficiently reduces hepatitis B virus cccDNA level in infected mice 脂质纳米粒包裹的 DOCK11-siRNA 可有效降低感染小鼠体内的乙型肝炎病毒 cccDNA 水平

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-06-24 DOI: 10.1016/j.omtm.2024.101289

Hikari Okada, Takeharu Sakamoto, Kouki Nio, Yingyi Li, Kazuyuki Kuroki, Saiho Sugimoto, Tetsuro Shimakami, Nobuhide Doi, Masao Honda, Motoharu Seiki, Shuichi Kaneko, Taro Yamashita

{"title":"Lipid nanoparticle-encapsulated DOCK11-siRNA efficiently reduces hepatitis B virus cccDNA level in infected mice","authors":"Hikari Okada, Takeharu Sakamoto, Kouki Nio, Yingyi Li, Kazuyuki Kuroki, Saiho Sugimoto, Tetsuro Shimakami, Nobuhide Doi, Masao Honda, Motoharu Seiki, Shuichi Kaneko, Taro Yamashita","doi":"10.1016/j.omtm.2024.101289","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101289","url":null,"abstract":"The hepatitis B virus (HBV) infects many people worldwide. As HBV infection frequently leads to liver fibrosis and carcinogenesis, developing anti-HBV therapeutic drugs is urgent. Therapeutic drugs for preventing covalently closed circular DNA (cccDNA) production, which can eliminate HBV infection, are unavailable. The host factor dedicator of cytokinesis 11 (DOCK11) is involved in the synthesis and maintenance of HBV cccDNA . However, the effectiveness of DOCK11 as a target for the elimination of HBV cccDNA remains unclear. In this study, we assess whether DOCK11 inhibitors suppress HBV cccDNA production in mouse models of HBV infection. The tocopherol-conjugate hetero gapmer, a DNA/RNA duplex of gapmer/complementary RNA targeting the DOCK11 sequence, partially reduces the expression of , but not that of HBV cccDNA, in the livers of HBV-infected human hepatocyte chimeric mice, along with weight loss and decreased serum human albumin levels. Lipid nanoparticle-encapsulated chemically modified siRNAs specific for suppress expression and decrease HBV cccDNA levels without adverse effects in the mice. Therefore, nucleic acid-based drugs targeting DOCK11 in hepatocytes are potentially effective anti-HBV therapeutics that can reduce HBV cccDNA levels .","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"27 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141551959","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

5′ Transgenes drive leaky expression of 3′ transgenes in Cre-inducible bi-cistronic vectors 5′ 转基因驱动 Cre 诱导的双组分载体中 3′ 转基因的泄漏表达

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-06-24 DOI: 10.1016/j.omtm.2024.101288

Yasuyuki Osanai, Yao Lulu Xing, Shinya Mochizuki, Kenta Kobayashi, Jihane Homman-Ludiye, Amali Cooray, Jasmine Poh, Ayumu Inutsuka, Nobuhiko Ohno, Tobias D. Merson

{"title":"5′ Transgenes drive leaky expression of 3′ transgenes in Cre-inducible bi-cistronic vectors","authors":"Yasuyuki Osanai, Yao Lulu Xing, Shinya Mochizuki, Kenta Kobayashi, Jihane Homman-Ludiye, Amali Cooray, Jasmine Poh, Ayumu Inutsuka, Nobuhiko Ohno, Tobias D. Merson","doi":"10.1016/j.omtm.2024.101288","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101288","url":null,"abstract":"Molecular cloning techniques enabling contemporaneous expression of two or more protein-coding sequences provide an invaluable tool for understanding the molecular regulation of cellular functions. The Cre-lox system is used for inducing the expression of recombinant proteins encoded within a bi-/poly-cistronic cassette. However, leak expression of transgenes is often observed in the absence of Cre recombinase activity, compromising the utility of this approach. To investigate the mechanism of leak expression, we generated Cre-inducible bi-cistronic vectors to monitor the expression of transgenes positioned either 5′ or 3′ of a 2A peptide or internal ribosomal entry site (IRES) sequence. Cells transfected with these bi-cistronic vectors exhibited Cre-independent leak expression specifically of transgenes positioned 3′ of the 2A peptide or IRES sequence. Similarly, AAV- vectors encoding bi-cistronic cassettes or fusion proteins revealed the selective Cre-independent leak expression of transgenes positioned at the 3′ end of the open reading frame. Our data demonstrate that 5′ transgenes confer promoter-like activity that drives the expression of 3′ transgenes. An additional STOP cassette between the 2A sequence and 3′ transgene dramatically decreased Cre-independent transgene expression. Our findings highlight the need for appropriate experimental controls when using Cre-inducible bi-/poly-cistronic constructs and inform improved design of vectors for more tightly regulated inducible transgene expression.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"88 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141553040","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

Evaluation of anti-vector immune responses to adenovirus-mediated lung gene therapy and modulation by αCD20 评估腺病毒介导的肺部基因治疗的抗载体免疫反应以及αCD20的调节作用

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-06-24 DOI: 10.1016/j.omtm.2024.101286

Robert D.E. Clark, Felix Rabito, Ferris T. Munyonho, T. Parks Remcho, Jay K. Kolls

{"title":"Evaluation of anti-vector immune responses to adenovirus-mediated lung gene therapy and modulation by αCD20","authors":"Robert D.E. Clark, Felix Rabito, Ferris T. Munyonho, T. Parks Remcho, Jay K. Kolls","doi":"10.1016/j.omtm.2024.101286","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101286","url":null,"abstract":"Although the last decade has seen tremendous progress in drugs that treat cystic fibrosis (CF) due to mutations that lead to protein misfolding, there are approximately 8%–10% of subjects with mutations that result in no significant CFTR protein expression demonstrating the need for gene editing or gene replacement with inhaled mRNA or vector-based approaches. A limitation for vector-based approaches is the formation of neutralizing humoral responses. Given that αCD20 has been used to manage post-transplant lymphoproliferative disease in CF subjects with lung transplants, we studied the ability of αCD20 to module both T and B cell responses in the lung to one of the most immunogenic vectors, E1-deleted adenovirus serotype 5. We found that αCD20 significantly blocked luminal antibody responses and efficiently permitted re-dosing. αCD20 had more limited impact on the T cell compartment, but reduced tissue resident memory T cell responses in bronchoalveolar lavage fluid. Taken together, these pre-clinical studies suggest that αCD20 could be re-purposed for lung gene therapy protocols to permit re-dosing.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"69 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141553041","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

Consensus-driven target product profiles for curative sickle cell disease gene therapies 治疗镰状细胞病基因疗法的共识驱动目标产品图谱

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-06-22 DOI: 10.1016/j.omtm.2024.101287

Daima Bukini, Julie Makani, Joseph McCune, Dennis Lee, Cathy Bansbach, Serena De Vita, Dominic Kemps, Elianna Amin, Jonathan Spector, John Tisdale

{"title":"Consensus-driven target product profiles for curative sickle cell disease gene therapies","authors":"Daima Bukini, Julie Makani, Joseph McCune, Dennis Lee, Cathy Bansbach, Serena De Vita, Dominic Kemps, Elianna Amin, Jonathan Spector, John Tisdale","doi":"10.1016/j.omtm.2024.101287","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101287","url":null,"abstract":"Therapeutic innovation to address sickle cell disease (SCD) is at an historical apex, characterized by a drug discovery, development, and commercialization landscape that includes potentially curative gene therapies. Given the wide geographic distribution of SCD, with a major presence in Africa, it is imperative that new medicines are designed to meet the specific needs of persons with SCD everywhere. Target product profiles (TPPs) detail the desired attributes of new medicines and serve as a guide for drug developers. To support research efforts for curative treatments for SCD, we mobilized a large multi-disciplinary expert group to generate consensus-driven TPPs for and SCD gene therapies, utilizing a modified Delphi methodology supplemented with virtual workshops. The main findings are TPPs that describe 20 minimal and optimal criteria for novel gene therapy products in categories of scope (3 criteria), performance/safety (11 criteria), manufacturing (4 criteria), and administration (2 criteria). TPPs for and products differed in some performance/safety criteria and all criteria pertaining to manufacturing and administration. These outputs will ideally support development of durable treatments that are safe, efficacious, and practical for persons with SCD in global settings.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"32 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141551962","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

Temporal insights into molecular and cellular responses during rAAV production in HEK293T cells 对 HEK293T 细胞中 rAAV 生产过程中分子和细胞反应的时空洞察

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-06-08 DOI: 10.1016/j.omtm.2024.101278

Alok Tanala Patra, Evan Tan, Yee Jiun Kok, Say Kong Ng, Xuezhi Bi

{"title":"Temporal insights into molecular and cellular responses during rAAV production in HEK293T cells","authors":"Alok Tanala Patra, Evan Tan, Yee Jiun Kok, Say Kong Ng, Xuezhi Bi","doi":"10.1016/j.omtm.2024.101278","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101278","url":null,"abstract":"The gene therapy field seeks cost-effective, large-scale production of recombinant adeno-associated virus (rAAV) vectors for high-dosage therapeutic applications. Although strategies like suspension cell culture and transfection optimization have shown moderate success, challenges persist for large-scale applications. To unravel molecular and cellular mechanisms influencing rAAV production, we conducted an SWATH-MS proteomic analysis of HEK293T cells transfected using standard, sub-optimal, and optimal conditions. Gene Ontology and pathway analysis revealed significant protein expression variations, particularly in processes related to cellular homeostasis, metabolic regulation, vesicular transport, ribosomal biogenesis, and cellular proliferation under optimal transfection conditions. This resulted in a 50% increase in rAAV titer compared with the standard protocol. Additionally, we identified modifications in host cell proteins crucial for AAV mRNA stability and gene translation, particularly regarding AAV capsid transcripts under optimal transfection conditions. Our study identified 124 host proteins associated with AAV replication and assembly, each exhibiting distinct expression pattern throughout rAAV production stages in optimal transfection condition. This investigation sheds light on the cellular mechanisms involved in rAAV production in HEK293T cells and proposes promising avenues for further enhancing rAAV titer during production.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"18 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141529693","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

Global seroprevalence of neutralizing antibodies against adeno-associated virus serotypes used for human gene therapies 用于人类基因疗法的腺相关病毒血清型中和抗体的全球流行率

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-05-29 DOI: 10.1016/j.omtm.2024.101273

Amit Chhabra, George Bashirians, Christos J. Petropoulos, Terri Wrin, Yuvika Paliwal, Peter V. Henstock, Suryanarayan Somanathan, Candida da Fonseca Pereira, Ian Winburn, John E.J. Rasko

{"title":"Global seroprevalence of neutralizing antibodies against adeno-associated virus serotypes used for human gene therapies","authors":"Amit Chhabra, George Bashirians, Christos J. Petropoulos, Terri Wrin, Yuvika Paliwal, Peter V. Henstock, Suryanarayan Somanathan, Candida da Fonseca Pereira, Ian Winburn, John E.J. Rasko","doi":"10.1016/j.omtm.2024.101273","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101273","url":null,"abstract":"Adeno-associated virus (AAV) vectors are promising gene therapy candidates, but pre-existing anti-AAV neutralizing antibodies (NAbs) pose a significant challenge to successful gene delivery. Knowledge of NAb seroprevalence remains limited and inconsistent. We measured activity of NAbs against six clinically relevant AAV serotypes across 10 countries in adults ( = 502) and children ( = 50) using a highly sensitive transduction inhibition assay. NAb prevalence was generally highest for AAV1 and lowest for AAV5. There was considerable variability across countries and geographical regions. NAb prevalence increased with age and was higher in females, participants of Asian ethnicity, and participants in cancer trials. Co-prevalence was most frequently observed between AAV1 and AAV6 and less frequently between AAV5 and other AAVs. Machine learning analyses revealed a unique clustering of AAVs that differed from previous phylogenetic classifications. These results offer insights into the biological relationships between the immunogenicity of AAVs in humans beyond that observed previously using standard clades, which are based on linear capsid sequences. Our findings may inform improved vector design and facilitate the development of AAV vector-mediated clinical gene therapies.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"14 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141516443","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

siRNA-mediated reduction of a circulating protein in swine using lipid nanoparticles 利用脂质纳米颗粒以 siRNA 为媒介减少猪体内的一种循环蛋白

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-04-27 DOI: 10.1016/j.omtm.2024.101258

Massimo F. Cau, Francesca Ferraresso, Monica Seadler, Katherine Badior, Youjie Zhang, Laura M. Ketelboeter, Geoffrey Rodriguez, Taylor Chen, Matteo Ferraresso, Amanda Wietrzny, Madelaine Robertson, Amber Haugen, Pieter R. Cullis, Marc de Moya, Mitchell Dyer, Christian J. Kastrup

{"title":"siRNA-mediated reduction of a circulating protein in swine using lipid nanoparticles","authors":"Massimo F. Cau, Francesca Ferraresso, Monica Seadler, Katherine Badior, Youjie Zhang, Laura M. Ketelboeter, Geoffrey Rodriguez, Taylor Chen, Matteo Ferraresso, Amanda Wietrzny, Madelaine Robertson, Amber Haugen, Pieter R. Cullis, Marc de Moya, Mitchell Dyer, Christian J. Kastrup","doi":"10.1016/j.omtm.2024.101258","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101258","url":null,"abstract":"Genetic manipulation of animal models is a fundamental research tool in biology and medicine but is challenging in large animals. In rodents, models can be readily developed by knocking out genes in embryonic stem cells or by knocking down genes through delivery of nucleic acids. Swine are a preferred animal model for studying the cardiovascular and immune systems, but there are limited strategies for genetic manipulation. Lipid nanoparticles (LNPs) efficiently deliver small interfering RNA (siRNA) to knock down circulating proteins, but swine are sensitive to LNP-induced complement activation-related pseudoallergy (CARPA). We hypothesized that appropriately administering optimized siRNA-LNPs could knock down circulating levels of plasminogen, a blood protein synthesized in the liver. siRNA-LNPs against plasminogen (siPLG) reduced plasma plasminogen protein and hepatic plasminogen mRNA levels to below 5% of baseline values. Functional assays showed that reducing plasminogen levels modulated systemic blood coagulation. Clinical signs of CARPA were not observed, and occasional mild and transient hepatotoxicity was present in siPLG-treated animals at 5 h post-infusion, which returned to baseline by 7 days. These findings advance siRNA-LNPs in swine models, enabling genetic engineering of blood and hepatic proteins, which can likely expand to proteins in other tissues in the future.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"21 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140883094","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

Development of novel lipoplex formulation methodologies to improve large-scale transient transfection for lentiviral vector manufacture 开发新型脂质体配制方法，改进慢病毒载体的大规模瞬时转染生产

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-04-26 DOI: 10.1016/j.omtm.2024.101260

Thomas Williams-Fegredo, Lee Davies, Carol Knevelman, Kyriacos Mitrophanous, James Miskin, Qasim A. Rafiq

{"title":"Development of novel lipoplex formulation methodologies to improve large-scale transient transfection for lentiviral vector manufacture","authors":"Thomas Williams-Fegredo, Lee Davies, Carol Knevelman, Kyriacos Mitrophanous, James Miskin, Qasim A. Rafiq","doi":"10.1016/j.omtm.2024.101260","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101260","url":null,"abstract":"Large-scale transient transfection has advanced significantly over the last 20 years, enabling the effective production of a diverse range of biopharmaceutical products, including viral vectors. However, a number of challenges specifically related to transfection reagent stability and transfection complex preparation times remain. New developments and improved transfection technologies are required to ensure that transient gene expression-based bioprocesses can meet the growing demand for viral vectors. In this paper, we demonstrate that the growth of cationic lipid-based liposomes, an essential step in many cationic lipid-based transfection processes, can be controlled through adoption of low pH (pH 6.40 to pH 6.75) and in low salt concentration (0.2× PBS) formulations, facilitating improved control over the nanoparticle growth kinetics and enhancing particle stability. Such complexes retain the ability to facilitate efficient transfection for prolonged periods compared with standard preparation methodologies. These findings have significant industrial applications for the large-scale manufacture of lentiviral vectors for two principal reasons. First, the alternative preparation strategy enables longer liposome incubation times to be used, facilitating effective control in a good manufacturing practices setting. Second, the improvement in particle stability facilitates the setting of wider process operating ranges, which will significantly improve process robustness and maximise batch-to-batch control and product consistency.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"14 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-04-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140883138","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