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: 10544091
• 负责人: Judith K Gwathmey
• 金额: $ 58.48万
• 依托单位: RUTGERS BIOMEDICAL AND HEALTH SCIENCES
• 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 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10544091
• 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 Death Induction; 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; Genes; 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; Mitochondria; Molecular; Mus; Muscle Cells; Muscular dystrophy cardiomyopathy; Myocardial; Myocardial Infarction; Myocardial Stunning; Myocardial dysfunction; Oxidative Stress; Participant; Pathology; Pathway interactions; Patients; Persons; Play; Predisposition; Prevention; Preventive therapy; Proteins; Reaction; Regulation; Reperfusion Injury; Reperfusion Therapy; Reporting; Role; Running; SLC11A2 gene; Sickle Cell Anemia; Signal Pathway; Signal Transduction; Testing; United States; Volatilization; Western Blotting; Woodchuck; beta Thalassemia; calcium uniporter; 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).

概括 在美国，大约有一个人每30秒死于心脏病。约150万 美国人每年死于心肌梗塞。临床上的遗传疾病（例如遗传 血色素沉着病）和反复输血（根据镰状细胞贫血和β地中海贫血的要求） 已知会导致铁超载心肌病在心脏中引起铁的积累 死亡。最近有报道称，大多数95％的患者发生了心肌病的扩张 在小鼠模型中，杜钦肌营养不良和铁水平升高。此外，铁水平是 缺血后，再灌注后，已知在心脏中升高。然而，基础 与Fe相关的心脏毒性涉及的机制尚不清楚。钙和铁都知道 在心脏中发挥重要的细胞作用。细胞表现出对钙和钙调节的显着依赖 铁浓度。任何一个离子的细胞失调都会导致收缩和舒张功能障碍，并且 最终的心肌病。细胞钙和/或细胞铁的正常稳态的丧失或破坏 浓度不仅会引起直接心肌心脏毒性，而且还会导致心肌丧失 兴奋性和异常激发收缩耦合。我们建议钙与 铁组合在高度心​​毒性细胞环境中产生。我们认为铁的存在会导致 在细胞死亡中，通过未充分的途径，即心脏的毛细杀来导致心肌病 作为缺血再灌注损伤。此外，我们提出了心肌惊人之间的类似联系 缺血再灌注的短期部分是由于相同的交叉谈话导致了部分可逆的 心肌收缩功能的降低。连接钙和铁信号的传导是线粒体 CA Uniporter（MCU）和瞬态受体电位规范通道的激活。我们表明铁 可以调节TRPC离子通道函数。我们的初步数据表明，TRPC通道直接 通过铁激活。重要的是，TRPC的激活与钙悖论损伤和钙后有关 心肌梗塞重塑。我们的目的是证明钙和铁都不是被动的 参与者参与细胞过程，但是当力加入时会导致心脏的收缩和舒张期衰竭， 心脏毒素，共同可以预测人类的寿命降低。我们将证明这是 细胞舒张性钙和线粒体钙，用铁定义细胞死亡和心肌功能 加载中。我们假设MCU解释了线粒体铁的超负荷和相互作用（或 crosstalk）之间的舒张钙升高和线粒体铁的增加导致高度挥发性和 心脏毒性通过铁毒性导致心脏细胞死亡的心脏毒性环境导致心肌病和 缺血再灌注损伤。在心脏上，许多方面都有许多有关铁凋亡的领域。 心脏中铁铁作用的主要驱动因素和途径因生物学环境而异。总之， 有很多可预见的机会来阐明触发器和激活的途径 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Duchenne在 肌肉营养不良的心肌病。我们将使用已证明的木屑来受到保护 从缺血再灌注损伤作为识别可以针对的新型抗肿瘤途径的工具 治疗和/或预防疗法。我们将追求以下目标。目标1：确定 FE中的MCU和TRPC在分离的心肌和体内诱导心脏功能障碍。 Sub-aim 1-1：我们将在体外演示和体内MCU是否介导Fe摄取和CA失调 与Fe诱导的心脏毒性有关。我们将确认MITO FE负载由MCU介导。这 Fe处理对MITO功能，氧化应激和MCU作用的影响将被定义。 Sub-aim 1-2：我们 将证明Fe诱导的TRPC激活以及体外（急性）和心脏功能障碍的关系 体内（Fe-CM）。 Sub-aim 1-3：从子-IAMS 1-1和1-2派生的数据将用于填充计算机 E-C-M耦合和模拟的模型与FE效应的结合运行。目标2：确定途径 在I/R损伤期间，在Fe-CM，DMD-CM和Woodchucks中涉及铁铁作用。 Sub-aim 2-1：我们将 确定通过MCU吸收MITO FE是否在FE载荷并测试其他已知的已知中发挥作用 铁凋亡的诱导剂。我们将确定ROS和TRPC在体外和体内的作用，并通过Fe负载和 i/r。 Sub-aim 2-2：在DMD-CM心脏中，我们将确定与铁毒相关基因和蛋白质的重塑， 评估生物标志物，并在体外测试对氟凋亡诱导剂的敏感性。我们将测试各种铁铁作用 途径并尝试通过抑制体内的铁毒性来减轻DMD-CM。 Sub-aim 2-3：我们将获得小说 在I/R期间（体外和体内）中的木屑保护机制的洞察力。