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

项目摘要 尽管在心血管疾病的诊断和治疗方面取得了进展，但心力衰竭的患病率仍然很低。 继续以流行病的速度增长。在应对各种损伤时，心脏会经历一个复杂的过程， 重塑，以胎儿表达程序的上调和代谢调控的相应转换为标志。 活动到胎儿样状态，与心肌病的发展密切相关。过程中 在K 08的初始资助期内，申请人接受了基于技术质谱的代谢组学培训， 计算科学，以及它们在询问培养细胞中的底物利用方面的集体应用， 器官水平生理学的代谢测量和代谢酶表达的变化。此外，申请人 已经应用这些方法来表征代谢重塑，发生在发病过程中， 心力衰竭，识别心脏中以前未被识别的代谢活动。这些代谢研究 然而，仅限于心肌损伤的啮齿动物模型，并且尚不清楚这些损伤的程度如何。 模型概括了人类心力衰竭的发展，特别是考虑到心脏疾病的关键差异， 大小、生理学和生物能量学之间的关系。发展和 人类诱导多能干细胞（iPSC）技术的进步已经改变了人类的研究， 疾病人iPSC在体外可容易地分化成心室心肌细胞（iPSC-CM）， 并且已经发现在体内概括了许多遗传学上的人类表型 心肌病，钙处理受损，力产生减少， 代谢胎儿基因程序。在本R 03提案中，申请人现在的目标是在初始K 08的基础上进行扩展 授予，并将全面的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具有序列- 经证实的家族性心肌病（扩张型和肥厚型心肌病），申请人将 通过最先进的液体， 色谱-质谱法。这些拟议中的研究将对代谢 人类心肌病的基础，区分收缩活动受损的新代谢物生物标志物，以及 确定代谢途径服从药理学和饮食干预，以及将形成 为申请人过渡到一个独立的，R 01资助的研究者奠定了基础。

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.