Developing CRISPR-based therapies for hypertrophic cardiomyopathy using human cardiomyocytes in a dish

肥厚型心肌病（HCM）是一种常见的心肌遗传性疾病，预计全球约有1/500的患者。大多数现有的治疗寻求改善症状，而不是解决潜在的疾病病理机制。该项目的目的是使用来自人类干细胞的心肌细胞（iPSC-CMs）作为模型系统来研究与HCM相关的遗传变异并开发治愈性遗传疗法。该项目的重点是开发基于CRISPR-Cas平台的新型基因编辑策略，以纠正驱动HCM的遗传病理机制，旨在全面恢复心脏受影响心肌细胞的健康收缩功能并逆转HCM表型。这项工作还将评估向心肌细胞提供CRISPR-Cas编辑机制的新技术。除了测试基因编辑策略，iPSC-CM平台还将用于评估与HCM相关的先前未表征的遗传变异的致病性，并提高我们对HCM驱动机制的理解。使用iPSC-CM需要在细胞培养、基本分子生物学技术和荧光显微镜方面进行培训。生成用于研究新型遗传变异的细胞系需要使用CRISPR-Cas9进行基因编辑的培训。开发和评估基因疗法将需要在生物信息学和转录组学方面进行额外的培训，以设计编辑策略并评估其疗效。该项目还将利用基于3D iPSC-CM的模型，试图在更类似于人类心脏中发现的成人心肌的心脏组织中建立疾病模型。这将需要在开发人类心脏类器官和工程心脏组织方面进行培训。

Hypertrophic cardiomyopathy (HCM) is a common genetic disease of the cardiac muscle, predicted to affect approximately 1 in 500 worldwide. Most existing treatments seek to ameliorate symptoms, rather than addressing the underlying disease pathomechanisms. The aim of this project is to use cardiomyocytes derived from human stem cells (iPSC-CMs) as a model system to study genetic variants associated with HCM and develop curative genetic therapies. The focus of this project is to develop novel gene editing strategies based on the CRISPR-Cas platform to correct the genetic pathomechanisms that drive HCM, with the aim of comprehensively restoring healthy contractile function in the affected cardiomyocytes of the heart and reversing the HCM phenotype. This work will also evaluate novel technologies to deliver CRISPR-Cas editing machinery to cardiomyocytes. Alongside testing gene editing strategies, the iPSC-CM platform will be used to evaluate the pathogenicity of previously-uncharacterised genetic variants associated with HCM, and to improve our understanding of the mechanisms that drive HCM. Working with iPSC-CMs requires training in cell culture, basic molecular biology techniques and fluorescence microscopy. Generating cell lines for studying novel genetic variants requires training in the use of CRISPR-Cas9 for gene editing. Developing and evaluating genetic therapies will require additional training in bioinformatics and transcriptomics to design editing strategies and assess their efficacy. This project will also utilise 3D iPSC-CM-based models in an attempt to model disease in a cardiac tissue more akin to the adult myocardium found in human hearts. This will require training in the development of human cardiac organoids and engineered heart tissues.