Molecular Therapy-Methods & Clinical Development最新文献

Non-encapsidated miRNA contaminants found in AAV preparations 在 AAV 制剂中发现的非包囊 miRNA 杂质

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-20 DOI: 10.1016/j.omtm.2024.101336

Mark A. Brimble, Stephen M. Winston

引用次数: 0

A novel AAV9-dual microRNA- vector targeting GRIK2 in the hippocampus as a treatment for mesial temporal lobe epilepsy 以海马中的 GRIK2 为靶点的新型 AAV9 双 microRNA 载体可用于治疗中位颞叶癫痫病

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-16 DOI: 10.1016/j.omtm.2024.101342

Stéphane J. Baudouin, April Giles, Nick Pearson, Severine Deforges, Chenxia He, Céline Boileau, Nicolas Partouche, Andreas Borta, Justine Gautron, Morgane Wartel, Irena Bočkaj, Didier Scavarda, Fabrice Bartolomei, Guillaume Penchet, Jérôme Aupy, Jennifer Sims, Jared Smith, Andrew Mercer, Olivier Danos, Christophe Mulle, Richard Porter

{"title":"A novel AAV9-dual microRNA- vector targeting GRIK2 in the hippocampus as a treatment for mesial temporal lobe epilepsy","authors":"Stéphane J. Baudouin, April Giles, Nick Pearson, Severine Deforges, Chenxia He, Céline Boileau, Nicolas Partouche, Andreas Borta, Justine Gautron, Morgane Wartel, Irena Bočkaj, Didier Scavarda, Fabrice Bartolomei, Guillaume Penchet, Jérôme Aupy, Jennifer Sims, Jared Smith, Andrew Mercer, Olivier Danos, Christophe Mulle, Richard Porter","doi":"10.1016/j.omtm.2024.101342","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101342","url":null,"abstract":"Mesial temporal lobe epilepsy (mTLE) is the most prevalent type of epilepsy in adults. First and subsequent generations of anti-epileptic therapy regimens fail to decrease seizures in a large number of patients suffering from mTLE, leaving surgical ablation of part of the hippocampus as the only therapeutic option to potentially reach seizure freedom. GluK2 has recently been identified as a promising target for the treatment of mTLE using gene therapy. Here, we engineered an adeno-associated virus serotype 9 vector expressing a cluster of two synthetic microRNAs (miRNAs), expressed from the human synapsin promoter, that target GRIK2 mRNA. Intra-hippocampal delivery of this vector in a mouse model of mTLE significantly reduced GRIK2 expression and daily seizure frequency. This treatment also improved the animals' health, reduced their anxiety, and restored working memory. Focal administration of the vector to the hippocampus of cynomolgus monkeys in GLP toxicology studies led to the selective transduction of hippocampal neurons with little exposure elsewhere in the brain and no transduction outside the central nervous system. Expression of miRNAs in hippocampal neurons resulted in substantially decreased GRIK2 mRNA expression. These data suggest that the intra-hippocampal delivery of a GMP-grade AAV9 coding for a synthetic miRNAs targeting GRIK2 is a promising treatment strategy for mTLE.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"15 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252865","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

Optimal different adeno-associated virus capsid/promoter combinations to target specific cell types in the common marmoset cerebral cortex 针对普通狨猴大脑皮层特定细胞类型的最佳腺相关病毒壳/启动子组合

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-13 DOI: 10.1016/j.omtm.2024.101337

Yasunori Matsuzaki, Yuuki Fukai, Ayumu Konno, Hirokazu Hirai

{"title":"Optimal different adeno-associated virus capsid/promoter combinations to target specific cell types in the common marmoset cerebral cortex","authors":"Yasunori Matsuzaki, Yuuki Fukai, Ayumu Konno, Hirokazu Hirai","doi":"10.1016/j.omtm.2024.101337","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101337","url":null,"abstract":"To achieve cell type-specific gene expression, using target cell type-tropic different adeno-associated virus (AAV) capsids is advantageous. However, their tropism across brain cell types in nonhuman primates has not been fully elucidated. We assessed the tropism of nine AAV serotype capsids (AAV1, 2, 5, 6, 7, 8, 9, rh10, and DJ) expressing enhanced green fluorescent protein (EGFP) by chicken β-actin hybrid (CBh) promoter in marmoset cerebral cortical cells. All nine AAV capsid vectors, especially AAV9 and AAVrh10, caused highly neuron-selective EGFP expression. Some AAV capsids, including AAV5, induced EGFP expression to a lesser extent in oligodendrocytes. Different ubiquitous cytomegalovirus (CMV) and CMV early enhancer/chicken β actin (CAG) promoters exhibited similar neuron-predominant transgene expression. Conversely, all nine AAV capsid vectors with the astrocyte-specific hGFA(ABC1D) promoter selectively expressed EGFP in astrocytes, except AAV5, which modestly expressed EGFP in oligodendrocytes. Oligodendrocyte-specific mouse myelin basic protein (mMBP) promoter in AAV5 vectors expressed EGFP in oligodendrocytes specifically and efficiently. The following are optimal combinations of capsids and promoters for cell type-specific expression: AAV9 or AAVrh10 and ubiquitous CBh or CMV promoter for neuron-specific transgene expression; AAV2 or AAV7 and hGFA(ABC1D) promoters for astrocyte-specific transgene expression; and AAV5 and mMBP promoters for oligodendrocyte-specific transgene expression.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"27 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142197239","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

An HPLC-SEC-based rapid quantification method for vesicular stomatitis virus particles to facilitate process development 基于 HPLC-SEC 的水泡性口炎病毒颗粒快速定量方法促进工艺开发

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-12 DOI: 10.1016/j.omtm.2024.101335

Adrian Schimek, Judy K.M. Ng, Ioannes Basbas, Fabian Martin, Dongyue Xin, David Saleh, Jürgen Hubbuch

引用次数: 0

DNA Contamination Within Recombinant Adeno Associated Virus (AAV) Preparations Correlates with Decreased CD34+ Cell Clonogenic Potential 重组腺相关病毒 (AAV) 制剂中的 DNA 污染与 CD34+ 细胞克隆生成潜能下降有关

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-12 DOI: 10.1016/j.omtm.2024.101334

Christopher Luthers, Sung-Min Ha, Annika Mittelhauser, Marco Morselli, Joseph D. Long, Caroline Y. Kuo, Zulema Romero, Donald B. Kohn

{"title":"DNA Contamination Within Recombinant Adeno Associated Virus (AAV) Preparations Correlates with Decreased CD34+ Cell Clonogenic Potential","authors":"Christopher Luthers, Sung-Min Ha, Annika Mittelhauser, Marco Morselli, Joseph D. Long, Caroline Y. Kuo, Zulema Romero, Donald B. Kohn","doi":"10.1016/j.omtm.2024.101334","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101334","url":null,"abstract":"Recombinant adeno-associated viruses (rAAV) are promising for applications in many genome editing techniques through their effectiveness as carriers of DNA homologous donors into primary hematopoietic stem and progenitor cells (HSPC) but have many outstanding concerns. Specifically, their biomanufacturing and the variety of factors that influence the quality and consistency of rAAV preps are in question. During the process of rAAV packaging, a cell line is transfected with several DNA plasmids that collectively encode all the necessary information to allow for viral packaging. Ideally, this process results in packaging of complete viral particles only containing rAAV genomes; however, this is not the case. Through this study, we were able to leverage Single-Stranded Virus (SSV)-seq, an NGS-based method to quantify all DNA species present within rAAV preps. From this, it was determined that much of the DNA within some rAAV preps is not vector-genome derived, and there is wide variability in the contamination by DNA across various preps. Furthermore, we demonstrate that transducing CD34+ hematopoietic stem and progenitor cells (HSPC) with preps with higher contaminating DNA resulted in decreased clonogenic potential, altered transcriptomic profiles, and decreased genomic editing. Collectively, this study characterized the effects of DNA contamination within rAAV preps on CD34+ HSPC cellular potential.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"68 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142197240","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

Assessment of adeno-associated virus purity by capillary electrophoresis-based western 用毛细管电泳法评估腺相关病毒的纯度

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-07 DOI: 10.1016/j.omtm.2024.101332

Julyana Acevedo, Yiling Bi, Jessica Gee, Santoshkumar L. Khatwani

引用次数: 0

Gene Therapy and kidney diseases 基因治疗与肾脏疾病

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-09-06 DOI: 10.1016/j.omtm.2024.101333

Nadia Tavakolidakhrabadi, Wen Y. Ding, Moin A. Saleem, Gavin I. Welsh, Carl May

引用次数: 0

Longitudinal characterization of sub-retinal pigment epithelium deposit formation in a primary porcine tissue culture model of dry age-related macular degeneration 干性老年性黄斑变性原代猪组织培养模型中视网膜下色素上皮沉积形成的纵向特征描述

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-08-31 DOI: 10.1016/j.omtm.2024.101331

Erika M. Shaw, Alexander J. Tate, Ramesh Periasamy, Daniel M. Lipinski

{"title":"Longitudinal characterization of sub-retinal pigment epithelium deposit formation in a primary porcine tissue culture model of dry age-related macular degeneration","authors":"Erika M. Shaw, Alexander J. Tate, Ramesh Periasamy, Daniel M. Lipinski","doi":"10.1016/j.omtm.2024.101331","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101331","url":null,"abstract":"Age-related macular degeneration (AMD) affects millions of individuals worldwide and is a leading cause of blindness in the elderly. In dry AMD, lipoproteinaceous deposits called drusen accumulate between the retinal pigment epithelium (RPE) and Bruch’s membrane, leading to impairment of oxygen and nutrient trafficking to the neural retina, and degeneration of the overlying photoreceptor cells. Owing to key differences in human and animal ocular anatomy and the slowly progressing nature of the disease, AMD is not easily modeled In this study, we further characterize a “drusen-in-a-dish” primary porcine RPE model system by employing vital lipid staining to monitor sub-RPE deposition over time in monolayers of cells cultured on porous transwell membranes. We demonstrate for the first time using a semi-automated image analysis pipeline that the number and size of sub-RPE deposits increases gradually but significantly over time and confirm that sub-RPE deposits grown in culture immunostain positive for multiple known components found in human drusen. As a result, we propose that drusen-in-a-dish cell culture models represent a high-throughput and cost-scalable alternative to animal models in which to study the pathobiology of drusen accumulation and may serve as useful tools for screening novel therapeutics aimed at treating dry AMD.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"19 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142197243","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

Synergy between Lactobacillus murinus and anti-PcrV antibody delivered in the airways to boost protection against Pseudomonas aeruginosa 鼠乳杆菌与在气道中递送的抗 PcrV 抗体之间的协同作用可增强对铜绿假单胞菌的抵抗力

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-08-30 DOI: 10.1016/j.omtm.2024.101330

Thomas Sécher, Mélanie Cortes, Chloé Boisseau, Marie-Thérèse Barba Goudiaby, Aubin Pitiot, Christelle Parent, Muriel Thomas, Nathalie Heuzé-Vourc’h

{"title":"Synergy between Lactobacillus murinus and anti-PcrV antibody delivered in the airways to boost protection against Pseudomonas aeruginosa","authors":"Thomas Sécher, Mélanie Cortes, Chloé Boisseau, Marie-Thérèse Barba Goudiaby, Aubin Pitiot, Christelle Parent, Muriel Thomas, Nathalie Heuzé-Vourc’h","doi":"10.1016/j.omtm.2024.101330","DOIUrl":"https://doi.org/10.1016/j.omtm.2024.101330","url":null,"abstract":"Therapeutic antibodies (Ab) have revolutionized the management of multiple illnesses including respiratory tract infections (RTIs). However, anti-infectious Ab displayed several limitations including antigen restrictiveness, narrowed therapeutic windows, and limited dose in the vicinity of the target when delivered by parenteral routes. Strategies enhancing further Ab-dependent containment of infection are currently needed. Here we showed that a combination of inhaled anti-infectious Ab and probiotics is an efficient formulation to protect against lung infection. Using a mouse model of Pseudomonas aeruginosa-induced pneumonia, we demonstrated a synergistic effect reducing both bacterial burden and pro-inflammatory response affording protection against primary and secondary infections. This is the first study showing that the local combination in the airways of anti-infective Ab and probiotics subverts suboptimal potency of Ab monotherapy and provides protection against respiratory pathogen.","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":"879 1","pages":""},"PeriodicalIF":4.7,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142197288","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

Resolving hidden subpopulations of filled AAVs by probing capsid integrity 通过探测囊壳完整性解析填充型 AAV 隐藏亚群

IF 4.7 2区医学

Molecular Therapy-Methods & Clinical Development Pub Date : 2024-08-29 DOI: 10.1016/j.omtm.2024.101322

Weida Wu, Georgios Katsikis

引用次数: 0