SCGE Disease Models Studies Supplement: Cardioediting Ttntvs in a humanized mouse model

SCGE 疾病模型研究补充：人源化小鼠模型中的心脏编辑 Ttntvs

• 批准号: 10619106
• 负责人: John Travis Hinson
• 金额: $ 23.24万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-09 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10619106
• 关键词: Animal Model; Cardiac Myocytes; Cardiovascular Diseases; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Complex; Data; Dependovirus; Development; Disease; Disease model; Enzymes; Functional disorder; Genetic Models; Goals; Heart failure; Human; Human Engineering; In Vitro; Knowledge; Modification; Molecular; Mus; Pathogenicity; Patients; Reading Frames; Reagent; Resources; Safety; Study models; System; Technology; Therapeutic; Toxic effect; Toxicology; Transcriptional Activation; Variant; cardiac tissue engineering; clinical development; connectin; efficacy evaluation; genetic variant; genome editing; genotoxicity; humanized mouse; immunogenicity; in vivo; mouse model; repaired; somatic cell gene editing; tool

ABSTRACT Somatic cell genome editing (SCGE) has remarkable promise to transform our therapeutic toolbox for heart failure due to Titin truncation variants (TTNtvs). However, despite having the knowledge of the pathogenic TTNtvs, their molecular consequences, and SCGE technologies including CRISPR genome editing and transcriptional activation to target TTNtvs, the lack of in vivo studies using humanized TTNtv animal models has hindered clinical development. There remain key knowledge gaps limiting SCGE development for TTNtvs including the lack of a systematic examination of the proportion of TTNtv alleles and cardiomyocytes that need to be corrected for clinical benefit, and the toxicology profile of SCGE enzymes and delivery systems including genotoxicity and immunogenicity. Our long-term goal is to develop SCGE for heart failure patients. While in vitro studies to evaluate the efficacy and safety of SCGE editors and delivery systems such as by utilizing human engineered heart tissues (EHTs) are important, they do not replace the need for animal model studies that recapitulate the complex pathophysiology of heart failure. Guided by our comprehensive preliminary data and unique reagents including SCGE components functionally validated in TTNtv EHTs, humanized Ttntv mice and cardiotropic adeno-associated viruses; our 1-year study will include two Specific Aims to determine efficacy and safety of 1) TTNtv reading frame repair and 2) TTN transcriptional activation in a humanized Ttntv mouse. Execution of these Aims will provide editing thresholds, safety, and toxicity for two SCGE approaches to treat TTNtvs that we expect will serve as a resource for the broader scientific community.

抽象的 体细胞基因组编辑（SCGE）具有重大的希望，可以改变我们的心脏治疗工具箱 由于TITIN截断变体（TTNTVS）而导致的故障。但是，尽管有病原体知识 TTNTV，它们的分子后果和SCGE技术，包括CRISPR基因组编辑和 转录激活靶向TTNTV，使用人源化TTNTV动物模型缺乏体内研究 阻碍临床发展。仍然存在关键的知识差距，限制了SCGE开发的TTNTVS 包括缺乏对需要的TTNTV等位基因和心肌细胞比例的系统检查 要纠正临床利益，以及SCGE酶和输送系统的毒理学概况 遗传毒性和免疫原性。我们的长期目标是为心力衰竭患者开发SCGE。在体外 评估SCGE编辑和交付系统（例如利用人类）的功效和安全性的研究 工程心脏组织（EHT）很重要，它们不能取代对动物模型研究的需求 概括心力衰竭的复杂病理生理学。以我们全面的初步数据为指导 独特的试剂，包括在TTNTV EHT，人性化TTNTV小鼠和 心形腺相关病毒；我们的一年研究将包括确定功效和 1）TTNTV阅读框架修复和2）人源化TTNTV小鼠中的TTN转录激活。 执行这些目标将为两种SCGE方法提供编辑阈值，安全性和毒性 我们期望的TTNTV将成为更广泛的科学界的资源。