Ferroptosis in the Heart: Iron Calcium Crosstalk and Compartmentalization

心脏铁死亡：铁钙串扰和区室化

• 批准号: 10364032
• 负责人: Judith K Gwathmey
• 金额: $ 60.35万
• 依托单位: RBHS-NEW JERSEY MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-25 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10364032
• 关键词: 3-Dimensional; Acute; Address; American; Animal Model; Biological; Biological Markers; Blood Transfusion; Calcium; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cardiotoxicity; Cause of Death; Cell Death; Cell physiology; Cells; Chelation Therapy; Clinical; Computer Models; Coupling; Cytosol; Data; Dependence; Development; Diastolic heart failure; Dilated Cardiomyopathy; Duchenne cardiomyopathy; Duchenne muscular dystrophy; Environment; Exhibits; Failure; Functional disorder; Gene Proteins; Genetic Diseases; Heart; Heart Diseases; Hereditary hemochromatosis; Homeostasis; Human; In Vitro; Injury; Intervention; Ion Channel; Ions; Iron; Iron Chelation; Iron Overload; Ischemia; Lead; Link; Lipid Peroxidation; Lipids; Longevity; Mediating; Medical Genetics; Mitochondria; Molecular; Mus; Muscle Cells; Myocardial; Myocardial Infarction; Myocardial Stunning; Myocardial dysfunction; Oxidative Stress; Participant; Pathology; Pathway interactions; Patients; Persons; Play; Predisposition; Prevention; Preventive therapy; Reaction; Regulation; Reperfusion Injury; Reperfusion Therapy; Reporting; Role; Running; SLC11A2 gene; Sickle Cell Anemia; Signal Pathway; Signal Transduction; Testing; United States; Western Blotting; Woodchuck; base; beta Thalassemia; calcium uniporter; crosslink; in vitro testing; in vivo; inhibitor; insight; lipidomics; mouse model; myocardial damage; myocardial injury; network models; novel; oxidation; receptor; simulation; targeted treatment; tool; uptake

Summary Approximately one person dies from heart disease every 30 seconds in the United States. About 1.5 million Americans die from myocardial infarction each year. Clinically, genetic disorders (e.g. hereditary hemochromatosis) and repeated blood transfusions (as required for sickle cell anemia and beta thalassemia) are known to cause Fe accumulation in the heart with iron overload cardiomyopathy being a major cause of death. It has been recently reported that dilated cardiomyopathy occurs in up to 95% of patients with Duchenne muscular dystrophy and that iron levels are elevated in mouse models. Furthermore, iron levels are known to be elevated in the heart after ischemia followed by reperfusion. Nevertheless, the underlying mechanism(s) involved in Fe associated cardiotoxicity remain unclear. Calcium and iron are both known to play vital cellular roles in the heart. Cells exhibit a remarkable dependence on keen regulation of calcium and iron concentrations. Cellular dysregulation of either ion can result in systolic and diastolic dysfunction and ultimately cardiomyopathy. Loss or disruption of normal homeostasis of cellular calcium and/or cellular iron concentrations can not only cause direct myocardial cardiotoxicity, but can also result in loss of myocardial excitability and abnormal excitation contraction coupling. We propose that a cross talk between calcium and iron combined results in a highly cardiotoxic cellular environment. We posit that the presence of iron can result in cell death via an underappreciated pathway, i.e. ferroptosis in the heart resulting in cardiomyopathy as well as ischemia reperfusion injury. Furthermore, we propose a similar link between myocardial stunning seen after brief periods of ischemia reperfusion to be in part due to the same cross talk resulting in a partially reversible reduction in myocardial systolic function. Linking the transport of calcium and iron signaling is the mitochondria Ca uniporter (mCU) and the activation of transient receptor potential canonical channels. We show that iron can regulate TRPC ion channel function. Our preliminary data have shown that TRPC channels are directly activated by iron. Importantly, activation of TRPCs has been implicated in calcium paradox injury and post- myocardial infarction remodeling. We aim to demonstrate that neither calcium nor iron are simply passive participants in cellular processes, but when forces are joined result in systolic and diastolic failure of the heart, cardiotoxcity, and together are predictive of a reduced lifespan in humans. We will demonstrate that it is cellular diastolic calcium and mitochondrial calcium that defines cell death and myocardial function with iron loading. We hypothesize that mCU accounts for mitochondrial iron overload and that an interaction (or crosstalk) between elevated diastolic calcium and increased mitochondrial iron results in a highly volatile and cardiotoxic environment that causes cardiac cell death via ferroptosis resulting in cardiomyopathy and ischemia reperfusion injury. The field of ferroptosis is nascent in many regards when it comes to the heart. The key drivers and pathways of ferroptosis in the heart differ depending on biological context. In summary, there is a wealth of foreseeable opportunities to elucidate both the trigger(s) and pathways activated that can result in ferroptosis and its role in various forms of cardiac cardiomyopathy and ischemia-reperfusion injury. Our preliminary studies have demonstrated ferroptosis in iron induced cardiomyopathy and Duchenne Muscular Dystrophy cardiomyopathy. We will use woodchucks that have been shown by us to be protected from ischemia reperfusion injury as a tool to identify novel anti-ferroptosis pathways that can be targeted for treatment and/or preventative therapies. We will pursue the following aims. Aim 1: Determine the role of mCU and TRPCs in Fe induced cardiac dysfunction at the level of the isolated myocyte and in vivo. Sub-aim 1-1: We will demonstrate in vitro and in vivo whether mCU mediated Fe uptake and Ca dysregulation are associated with Fe induced cardiac toxicity. We will confirm mito Fe loading is mediated by mCU. The effects of Fe treatment on mito function, oxidative stress and the role of mCU will be defined. Sub-aim 1-2: We will demonstrate Fe induced activation of TRPCs and the relationship to cardiac dysfunction in vitro (acute) and in vivo (Fe-CM). Sub-aim 1-3: Data derived from Sub-aims 1-1 and 1-2 will be used to populate a computer model of E-C-M coupling and simulations run with incorporation of Fe effects. Aim 2: To determine pathways involved in ferroptosis in Fe-CM, DMD-CM, and Woodchucks during I/R injury. Sub-aim 2-1: We will determine whether mito Fe uptake via mCU plays a role in ferroptosis with Fe loading and test other known inducers of ferroptosis. We will determine the role of ROS and TRPC in vitro and in vivo with Fe loading and I/R. Sub-aim 2-2: In DMD-CM hearts, we will determine remodeling of ferroptosis-related genes and proteins, evaluate biomarkers, and test the susceptibility to ferroptosis inducers in vitro. We will test various ferroptosis pathways and attempt to mitigate DMD-CM by inhibiting ferroptosis in vivo. Sub-aim 2-3: We will obtain novel insights into protective mechanism(s) in woodchucks during I/R (in vitro and in vivo).

总结 在美国，每30秒就有一人死于心脏病。大约150万 美国人每年死于心肌梗塞。临床上，遗传性疾病（如遗传性 血色素沉着症）和反复输血（如镰状细胞贫血和β地中海贫血所需） 已知会导致心脏中的铁积累，铁超载心肌病是心脏病的主要原因。 死亡最近有报道称，扩张型心肌病发生在高达95%的患者中， 杜氏肌营养不良症和铁水平在小鼠模型中升高。此外，铁含量 已知在缺血再灌注后心脏中升高。然而，潜在的 与铁相关的心脏毒性的机制仍不清楚。钙和铁都是已知的 在心脏中起着至关重要的细胞作用。细胞表现出对钙的强烈调节的显著依赖性， 铁浓度。任一离子的细胞失调可导致收缩和舒张功能障碍， 最后是心肌病。细胞钙和/或细胞铁的正常稳态丧失或破坏 浓度不仅可引起直接的心肌毒性，而且还可导致心肌细胞的损失。 兴奋性和异常兴奋收缩偶联。我们认为钙和钙离子之间的相互作用 铁结合导致高度心脏毒性的细胞环境。我们认为铁的存在会导致 通过一种未被充分认识的途径导致细胞死亡，即心脏中的铁凋亡也导致心肌病 缺血再灌注损伤。此外，我们提出了类似的联系心肌顿抑后， 短暂的缺血再灌注部分是由于相同的串扰，导致部分可逆的 心肌收缩功能下降。连接钙和铁信号的运输是线粒体 钙单向转运体（mCU）和瞬时受体电位经典通道的激活。我们发现铁 可调节TRPC离子通道功能。我们的初步数据表明，TRPC通道直接 由铁激活。重要的是，TRPC的激活与钙反常损伤和后钙反常损伤有关。 心肌梗死重构我们的目的是证明钙和铁都不是简单的被动的 细胞过程的参与者，但当力量联合起来导致心脏的收缩和舒张衰竭时， 心脏毒性，并共同预测人类寿命缩短。我们将证明， 细胞舒张期钙和线粒体钙，与铁一起定义细胞死亡和心肌功能 加载中我们假设mCU是线粒体铁超载的原因， 升高的舒张期钙和增加的线粒体铁之间的串扰）导致高挥发性的， 心脏毒性环境，通过铁凋亡引起心脏细胞死亡，导致心肌病， 缺血再灌注损伤当涉及到心脏时，铁下垂的领域在许多方面都是新生的。 心脏中铁凋亡的关键驱动因素和途径因生物学背景而异。总的来说， 有大量可预见的机会来阐明触发因素和激活的途径， 导致铁凋亡及其在各种形式的心肌病和缺血-再灌注损伤中的作用。 我们的初步研究表明，铁诱导的心肌病和杜氏病中的铁细胞凋亡 肌营养不良性心肌病。我们将使用土拨鼠，已表明我们受到保护 从缺血再灌注损伤作为一种工具，以确定新的抗铁凋亡途径，可以针对 治疗和/或预防性疗法。我们将追求以下目标。目标1：确定 铁中的mCU和TRPC在离体心肌细胞水平和体内均引起心功能障碍。 子目标1-1：我们将在体外和体内证明mCU是否介导Fe摄取和Ca失调 与铁诱导的心脏毒性有关。我们将证实线粒体铁负载是由mCU介导的。的 铁处理对线粒体功能、氧化应激和mCU的作用的影响将被定义。次级目标1-2：我们 将证明铁诱导的TRPC活化以及与体外（急性）心功能障碍的关系， 体内（Fe-CM）。次级目标1-3：从次级目标1-1和1-2获得的数据将用于输入计算机 模型的E-C-M耦合和模拟运行与铁的影响。目标2：确定途径 参与I/R损伤期间Fe-CM、DMD-CM和土拨鼠的铁凋亡。次级目标2-1：我们将 确定通过mCU的mito Fe摄取是否在Fe负荷的铁凋亡中起作用，并测试其他已知的 铁下垂的诱导剂。我们将确定ROS和TRPC在体外和体内的作用， I/R。子目标2-2：在DMD-CM心脏中，我们将确定铁蛋白沉积相关基因和蛋白的重塑， 评估生物标志物，并在体外测试对铁凋亡诱导剂的敏感性。我们将测试各种铁性下垂 途径，并试图通过抑制体内铁凋亡来缓解DMD-CM。子目标2-3：我们将获得新的 深入了解土拨鼠在I/R期间的保护机制（体外和体内）。