Genetic Pathways in Ceramide-Associated Lipotoxic Cardiomyopathy and Heart Failure

神经酰胺相关脂毒性心肌病和心力衰竭的遗传途径

• 批准号: 10311508
• 负责人: ROLF BODMER
• 金额: $ 48.75万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-10 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10311508
• 关键词: Address; Adult; Affect; Annexins; Apoptosis; Biological Models; Branched-Chain Amino Acids; Cardiac; Cardiac Myosins; Cardiomyopathies; Cardiovascular Diseases; Cardiovascular system; Categories; Cell Death; Cell Survival; Cell physiology; Ceramides; Clinical; Collaborations; DNA Damage; Data; Development; Diabetes Mellitus; Diet; Drosophila genus; Epidemic; Exhibits; FASN gene; Fatty acid glycerol esters; Fatty-acid synthase; Functional disorder; Genetic; Genetic Models; Glycolysis; Goals; Grant; Heart; Heart Abnormalities; Heart Diseases; Heart failure; Heat shock proteins; High Fat Diet; Homeostasis; Human; Knowledge; Letters; Lipids; Maintenance; Malignant Neoplasms; Mammalian Genetics; Mediating; Mediator of activation protein; Metabolic; Metabolic Control; Metabolic Pathway; Modeling; Molecular; Molecular Chaperones; Mus; Muscle; Myosin ATPase; Obesity; Ontology; Pathologic; Pathology; Pathway interactions; Physiology; Prevention; Proteins; RNA Interference; Regulation; Research; Research Personnel; Risk Factors; Role; Sarcomeres; Sphingolipids; Structure; Testing; Therapeutic Intervention; Triglycerides; United States; Woman; cardioprotection; cofactor; dietary; experimental study; fatty acid metabolism; fly; genetic regulatory protein; heart function; in vivo; induced pluripotent stem cell; induced pluripotent stem cell derived cardiomyocytes; insight; knock-down; lipid biosynthesis; men; mortality; novel; overexpression; overweight adults; prevent; response

Summary Heart disease is the leading cause of mortality in the United States in both men and women. Obesity is rapidly growing epidemic and a major risk factor for the development of cardiomyopathies and heart failure. The pathology of obesity-related cardiomyopathies is associated with abnormal cardiac accumulation of fat and lipotoxic metabolites. Preventing such lipotoxic effects is a potential avenue for therapeutic intervention in heart disease and heart failure. However, the molecular mechanisms of cardiac lipotoxicity remain elusive. In this proposal, we seek to address these gaps in knowledge by leveraging our established, robust Drosophila heart models of ceramide- and high-fat-diet (HFD)-induced lipotoxic cardiomyopathy (LCM), which exhibit LCM-like pathology observed in mammalian LCM, including compromised contractility and elevated accumulation of both triglyceride fat and sphingolipid ceramide with either diet. Drosophila is an extremely versatile genetic model system with highly conserved metabolic pathways and cardiac physiology, well suited to address fundamental mechanisms of LCM. Nevertheless, we will validate our findings in Drosophila in human cardiomyocytes that are derived from induced pluripotent stem cell (hiPSC-CMs) to gain further human relevant insights into the mechanisms of LCM (in collaboration with co-Investigator Dr. Alexandre Colas – see letter). In ceramide-protein trap experiments we identified 3 enriched ontologies of putative ceramide interacting proteins (180 putative overlapping mouse-human CIPs): (1) sarcomeric regulatory proteins (40 CIPs), including the myosin chaperone Uncoordinated-45b (Unc45b), (2) metabolic regulatory proteins (33 CIPs), including fatty acid synthase FASN, and (3) apoptosis & DNA damage response (DDR) proteins (21 CIPs). Indeed, found that cardiac Unc45 overexpression or FASN RNAi knockdown in the fly heart prevents ceramide-induced LCM. LCM- inducing and subthreshold-sensitizing ceramide and high fat treatments will be used to interrogate the above CIPs in participating in LCM. This will be the first step in determining in detail the mechanisms how ceramide interacts with these 3 classes of proteins and how that disrupts or protects cardiac function and homeostasis. The goal is to construct and test a myofibrillar maintenance/lipogenesis/ apoptosis-centric regulatory network weighted with novel CIPs, which we will experimentally evaluate further in our LCM models.

概括 心脏病是美国男性和女性死亡的主要原因。肥胖是 流行病迅速蔓延，是心肌病和心力衰竭发展的主要危险因素。这 肥胖相关心肌病的病理学与脂肪和脂肪的异常心脏积聚有关。 脂毒性代谢物。预防这种脂毒性作用是心脏治疗干预的潜在途径 疾病和心力衰竭。然而，心脏脂肪毒性的分子机制仍不清楚。 在本提案中，我们寻求通过利用我们既定的、强大的 神经酰胺和高脂饮食（HFD）诱导的脂毒性心肌病（LCM）的果蝇心脏模型， 表现出在哺乳动物 LCM 中观察到的类似 LCM 的病理学特征，包括收缩性受损和升高 无论哪种饮食，甘油三酯脂肪和鞘脂神经酰胺都会积累。果蝇是一种极其 具有高度保守的代谢途径和心脏生理学的多功能遗传模型系统，非常适合 解决 LCM 的基本机制。尽管如此，我们将在人类果蝇中验证我们的发现 源自诱导多能干细胞 (hiPSC-CM) 的心肌细胞，以获得进一步的人类相关性 对 LCM 机制的见解（与联合研究员 Alexandre Colas 博士合作 - 见信件）。 在神经酰胺-蛋白质陷阱实验中，我们确定了 3 种丰富的假定神经酰胺相互作用本体 蛋白质（180 个假定的重叠小鼠-人 CIP）：（1）肌节调节蛋白（40 个 CIP），包括 肌球蛋白伴侣 Uncooperative-45b (Unc45b)，(2) 代谢调节蛋白 (33 CIP)，包括脂肪 酸合酶 FASN，以及 (3) 细胞凋亡和 DNA 损伤反应 (DDR) 蛋白（21 个 CIP）。确实发现 果蝇心脏中 Unc45 过度表达或 FASN RNAi 敲除可预防神经酰胺诱导的 LCM。 LCM- 诱导和阈下敏化神经酰胺和高脂肪治疗将用于询问上述情况 CIP 参与 LCM。这将是详细确定神经酰胺如何发挥作用的机制的第一步 与这三类蛋白质相互作用，以及如何破坏或保护心脏功能和体内平衡。 目标是构建和测试以肌原纤维维持/脂肪生成/细胞凋亡为中心的调节网络 加权新的 CIP，我们将在 LCM 模型中进一步进行实验评估。