人诱导多能干细胞衍生心脏细胞外泌体治疗猪心肌梗死的机制研究

心肌梗死等心血管疾病是世界范围发病和死亡的主要原因。前期报道人诱导多能干细胞（hiPSCs）衍生的各种心脏细胞（hiPSC-CCs）分泌的外泌体（CC-Exo）可保护受损心肌和促进血管生长的作用，具有临床应用治疗心肌梗死的潜能。但是目前各类型CC-Exo组分、作用差异性、大动物安全性和疗效、作用机制等问题不清楚？我们已经成功从hiPSCs分化得到六种hiPSC-CCs（心肌、内皮、平滑肌、成纤维、心外膜、周细胞）; 分离提取了其外泌体，并在体内、体外初步观察到其保护作用。为了加快CC-Exo 临床转化，本课题旨在：1）体外研究不同类型CC-Exo的特征和成份谱，比较其保护作用；2）在猪心梗模型上确认主要类型CC-Exo的作用疗效和安全性，及其调控室性心律失常和钙稳态的作用机制；3）探索增强CC-Exo体内靶向性和稳定性的方法，从而为其临床治疗心梗建立基础。

Cardiovascular diseases such as myocardial infarction (MI) are the leading causes of mortality in China. Exosomes (CC-Exo) secreted from the human-induced pluripotent stem cell (hiPSC)-differentiated cardiac cells (hiPSC-CCs) were reported to have the potentiality to protect the injured cardiomyocyte and promote the vascular growth, which would be a candidate for the treatment against MI in the clinic. However, the components and effects of various types of CC-Exo, the safety and efficacy of specific types of CC-Exo in large animals, and the mechanisms for the cardio protection are still unclear. We have successfully differentiated various types of high-purity hiPSC-CCs (cardiomyocytes, endothelial cells, smooth muscle cells, fibroblasts, epicardial cells, and pericytes) from hiPSCs and isolated their exosomes. Moreover, the cardioprotective effects of these CC-Exo were observed in the pilot experiments in vitro and in vivo. In order to accelerate the clinical translation of CC-Exo, this project aims to: 1) investigate the characteristics and constituent profile of different types of CC-Exo, and compare the differences in cardio protection; 2) confirm the efficacy and safety of the specific types of CC-Exo in a porcine MI model; and focus on the mechanism investigation of CC-Exo in regulating porcine ventricular arrhythmia and maintaining myocyte intracellular calcium homeostasis; 3) explore the potential approaches to enhance the targeting and stability of CC-Exo in vivo.