Base Editing Gene Correction of Pathogenic MYH7 Mutations in Models of Hypertrophic Cardiomyopathy

肥厚型心肌病模型中致病性 MYH7 突变的碱基编辑基因校正

基本信息

批准号：
10733414
负责人：
Andreas C Chai
金额：
$ 4.56万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10733414
关键词：
ATP phosphohydrolase Affect Animal Model Arrhythmia CRISPR/Cas technology Cardiac Cardiac Myocytes DNA Sequence Alteration Dependovirus Development Disease Disease Progression Duchenne muscular dystrophy Echocardiography Energy consumption Foundations Gene Expression Genes Genetic Genomics Heart Heart Abnormalities Heart Diseases Heart failure High-Throughput Nucleotide Sequencing Histology Human Human Genome Hypertrophic Cardiomyopathy Hypertrophy Mediating Methods Microscopy Missense Mutation Modeling Motor Mus Mutation Myocardium Myopathy Myosin ATPase Myosin Heavy Chains Nucleotides Outcome Pathogenicity Pathologic Pathology Patients Persons Phenotype Plasmids Play Property Proteins Role Sarcomeres Therapeutic Thick Filament Tissues Transplantation United States Variant adeno-associated viral vector base editing base editor beta-Myosin clinically relevant design disease-causing mutation gene correction genome editing heart function human model induced pluripotent stem cell induced pluripotent stem cell derived cardiomyocytes laboratory experience molecular marker mouse model mutant novel novel therapeutic intervention prevent prime editing stem cell model sudden cardiac death therapeutic genome editing tool

项目摘要

PROJECT SUMMARY/ABSTRACT Hypertrophic cardiomyopathy (HCM), a disease of abnormal heart muscle thickening, is the most common form of genetic heart disease in the United States affecting upwards of 1 in 500 people. While HCM- causing mutations are found in various sarcomere protein-encoding genes, over one-third of all HCM-causing mutations occur in the Myosin Heavy Chain 7 (MYH7) gene, which encodes for beta-myosin heavy chain, a motor ATPase that incorporates into the thick filament of cardiac muscle and plays a major role in cardiac contraction. Missense mutations within MYH7 allow the incorporation of mutant myosin heads into cardiac sarcomeres, which leads to increased cardiomyocyte energy consumption, hypercontractility, and the initiation of the disease progression of HCM. For patients, complications of HCM include heart failure, arrhythmia, and sudden cardiac death. As HCM has no cure aside from transplant, there is an urgent need for novel therapeutic strategies. CRISPR-Cas9-mediated genome editing has emerged as an attractive method to correct and potentially cure genetically-based diseases. CRISPR-Cas9 and its variants, including base editing and prime editing, allow targeted genomic editing at specific loci. Base editing allows precise single nucleotide edits, which makes it an ideal therapeutic tool to correct the hundreds of documented HCM-causing missense mutations. Several studies have successfully demonstrated the use of adeno-associated virus (AAV)-mediated gene editing to modify disease-causing mutations in various tissues and rescue pathological phenotypes in small and large animal models. In particular, various studies have shown robust gene editing in the heart to treat the muscle disease Duchenne muscular dystrophy, raising the prospect of successful gene editing in the heart to treat other cardiac diseases. The Specific Aims of this study seek to demonstrate base editing as a cure for MYH7 mutations of hypertrophic cardiomyopathy in 1) human induced pluripotent stem cell-derived cardiomyocytes and 2) a mouse model of HCM. Both models of HCM contain the same pathogenic mutation commonly found in patients and the mouse model faithfully recapitulates the human phenotype of hypertrophic cardiomyopathy. The central hypothesis of this proposal is that AAV delivery of base editing components to each of these models will prevent HCM and revert established HCM. This proposal will assess gene editing on-target and off-target efficiencies and changes in cardiac function and pathology. Furthermore, these studies will develop and optimize a base editing strategy to cure hypertrophic cardiomyopathy and will lay a foundation for the development of gene editing therapies for the hundreds of other HCM-causing genetic mutations.

项目概要/摘要肥厚型心肌病（HCM）是一种心肌异常增厚的疾病，是最常见的心肌病。在美国，这是一种常见的遗传性心脏病，影响每 500 人中就有 1 人。而 HCM- 导致 HCM 的突变存在于各种肌节蛋白编码基因中，超过三分之一突变发生在肌球蛋白重链 7 (MYH7) 基因中，该基因编码 β-肌球蛋白重链，运动 ATP 酶，融入心肌粗丝，在心脏功能中发挥重要作用收缩。 MYH7 内的错义突变允许突变肌球蛋白头掺入心脏肌节，导致心肌细胞能量消耗增加、收缩力亢进和启动 HCM 的疾病进展。对于患者来说，HCM 的并发症包括心力衰竭、心律失常和心源性猝死。由于 HCM 除移植外无法治愈，因此迫切需要新的治疗方法治疗策略。 CRISPR-Cas9 介导的基因组编辑已成为一种有吸引力的方法来纠正和有可能治愈遗传性疾病。 CRISPR-Cas9 及其变体，包括碱基编辑和 Prime 编辑，允许在特定位点进行靶向基因组编辑。碱基编辑允许精确的单核苷酸编辑，这使其成为纠正数以百计的 HCM 引起错义的理想治疗工具突变。多项研究已成功证明使用腺相关病毒（AAV）介导的基因编辑修改各种组织中的致病突变并挽救病理表型小型和大型动物模型。特别是，各种研究表明心脏中强大的基因编辑治疗肌肉疾病杜氏肌营养不良症，提高基因编辑成功的前景心脏可治疗其他心脏疾病。本研究的具体目标旨在证明碱基编辑可以治愈 MYH7 突变 1) 人诱导多能干细胞来源的心肌细胞和 2) a 中的肥厚性心肌病 HCM 小鼠模型。两种 HCM 模型均含有患者中常见的相同致病突变小鼠模型忠实地再现了人类肥厚型心肌病的表型。这该提案的中心假设是 AAV 将碱基编辑组件传递给每个模型将预防 HCM 并恢复已建立的 HCM。该提案将评估基因编辑的靶向和脱靶心脏功能和病理学的效率和变化。此外，这些研究将发展并优化碱基编辑策略治疗肥厚型心肌病，为治疗肥厚型心肌病奠定基础针对数百种其他导致 HCM 的基因突变开发基因编辑疗法。