Mapping metabolic activities in stem cell-derived models of human cardiomyopathy

项目总结 尽管心血管疾病的诊断和治疗取得了进展，但心力衰竭的患病率 继续以流行病的速度上升。作为对各种损伤的反应，心脏经历了一个复杂的过程 重塑，以胎儿表达程序上调和相应的代谢转换为标志 活动到胎儿状态，与心肌病的发展密切相关。在整个过程中 最初的K08资助期申请人接受了基于质谱学的代谢组学技术培训，并 计算科学，以及它们在询问培养细胞中底物利用方面的集体应用， 器官水平生理的代谢测量，以及代谢酶表达的变化。此外，申请人 应用这些方法来表征在糖尿病发病机制中发生的代谢重塑。 心力衰竭，识别心脏中以前未被识别的代谢活动。这些新陈代谢研究， 然而，仅限于啮齿类动物的心肌损伤模型，目前尚不清楚这些损伤的程度。 模型概括了人类心力衰竭的发展过程，特别是考虑到心脏疾病的严重差异。 人和啮齿动物心肌细胞的大小、生理和生物能量学。中国的发展和 人类诱导多能干细胞(IPSC)技术的进步改变了人类的研究 疾病。人IPSCs在体外可以很容易地分化为心肌细胞 高效率并已被发现在体内概括了许多基因的人类表型 心肌病，钙处理受损，力量产生减少，并重新表达 代谢性胎儿基因程序。在这份R03提案中，申请人现在的目标是在最初的K08基础上延长 获奖，并将全面代谢组学应用于人类心肌病的研究，努力破译 人类心力衰竭的代谢基础。使用来自患有Sequence-Sequence-Sequence的患者的IPSC-CMS 经核实的家族性心肌病(扩张型和肥厚型心肌病)申请人将 通过最先进的液体全面定义心脏病理过程中发生的代谢变化 色质联用。这些拟议的研究将为新陈代谢提供巨大的洞察力 人类心肌病的基础，区分收缩活动受损的新代谢物生物标记物，以及 确定符合药物和饮食干预的代谢途径，以及将形成 为申请者过渡到独立的、R01资助的调查员的基金会。

PROJECT SUMMARY Despite advances in the diagnosis and treatment of cardiovascular disease, the prevalence of heart failure continues to rise at epidemic rates. In response to varied injuries, the heart undergoes a complex process of remodeling, marked by upregulation of a fetal expression program and corresponding switching of metabolic activity to a fetal-like state, in close association with the development of cardiomyopathy. Over the course of the initial K08 funding period the Applicant trained in technical mass spectrometry based metabolomics and computational sciences, and their collective application to interrogate substrate utilization in cells in culture, metabolic measures of organ level physiology, and shifts in metabolic enzyme expression. Moreover, Applicant has applied these approaches to characterize the metabolic remodeling that occurs during the pathogenesis of heart failure, identifying previously unrecognized metabolic activities in the heart. These metabolic studies, however, have been limited to rodent models of myocardial injury, and it remains unclear to what extend these models recapitulate the development of human heart failure, particularly given critical differences in cardiac size, physiology, and bioenergetics between human and rodent cardiomyocytes. The development and advance of human induced pluripotent stem cells (iPSC) technologies has transformed the study of human disease. Human iPSCs may be readily differentiated into ventricular cardiomyocytes (iPSC-CMs) in vitro at high efficiencies and have been found to recapitulate in vivo human phenotypes across a number of genetic cardiomyopathies, with impaired calcium handling, decreased force generation, and re-expression of the metabolic fetal gene program. In this R03 proposal, the Applicant now aims to extend upon the initial K08 award, and apply comprehensive metabolomics to the study of human cardiomyopathy, in an effort to decipher the metabolic underpinnings of human heart failure. Using iPSC-CMs derived from patients with sequence- verified familial cardiomyopathies (dilated and hypertrophic cardiomyopathy) the Applicant will comprehensively define the metabolic changes that occur with cardiac pathology through state-of-the-art liquid chromatography - mass spectrometry. These proposed studies will shed tremendous insight into the metabolic basis for human cardiomyopathy, distinguish new metabolite biomarkers for impaired contractile activity, and identify metabolic pathways amenable to pharmacologic and dietary intervention, as well as will form the foundation for the Applicant's transition to an independent, R01 funded investigator.