Ferroptosis in the Heart: Iron Calcium Crosstalk and Compartmentalization

心脏铁死亡：铁钙串扰和区室化

基本信息

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
项目类别：
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

项目摘要

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，大约每 30 秒就有一人死于心脏病。约150万每年都有美国人死于心肌梗塞。临床上，遗传性疾病（例如遗传性血色素沉着病）和反复输血（根据镰状细胞性贫血和β地中海贫血的需要）已知会导致铁在心脏中积聚，铁超负荷心肌病是铁过量心肌病的主要原因死亡。最近有报道称，高达 95% 的扩张型心肌病患者患有扩张型心肌病。杜氏肌营养不良症和小鼠模型中铁水平升高。此外，铁含量已知在缺血再灌注后心脏中升高。尽管如此，底层铁相关心脏毒性的机制尚不清楚。众所周知，钙和铁在心脏中发挥重要的细胞作用。细胞表现出对钙和细胞的敏锐调节的显着依赖性铁浓度。任何一种离子的细胞失调都会导致收缩和舒张功能障碍，最终导致心肌病。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心肌毒性，还可导致心肌损伤兴奋性与异常兴奋收缩耦合。我们建议钙和钙之间的串扰铁结合会导致高度心脏毒性的细胞环境。我们假设铁的存在会导致通过未被充分认识的途径导致细胞死亡，即心脏中的铁死亡也会导致心肌病如缺血再灌注损伤。此外，我们提出了心肌顿抑后观察到的类似联系短暂的缺血再灌注部分是由于相同的串扰导致部分可逆心肌收缩功能降低。连接钙和铁信号传导的是线粒体 Ca 单向转运蛋白 (mCU) 和瞬时受体潜在经典通道的激活。我们表明铁可以调节TRPC离子通道功能。我们的初步数据表明，TRPC 渠道直接由铁激活。重要的是，TRPC 的激活与钙悖论损伤和后遗症有关。心肌梗塞重构。我们的目标是证明钙和铁都不是简单的被动物质细胞过程的参与者，但当力量联合起来时会导致心脏的收缩和舒张衰竭，心脏毒性，并且共同预示着人类寿命的缩短。我们将证明它是细胞舒张钙和线粒体钙，用铁定义细胞死亡和心肌功能加载中。我们假设 mCU 是线粒体铁超载的原因，并且相互作用（或舒张期钙升高和线粒体铁增加之间的串扰）导致高度不稳定和心脏毒性环境，通过铁死亡导致心肌细胞死亡，导致心肌病和缺血再灌注损伤。当涉及心脏时，铁死亡领域在许多方面都处于新生阶段。心脏铁死亡的关键驱动因素和途径因生物学背景而异。总之，有大量可预见的机会来阐明可以激活的触发因素和途径导致铁死亡及其在各种形式的心肌病和缺血再灌注损伤中的作用。我们的初步研究已经证明铁诱导的心肌病和杜氏心肌病中存在铁死亡肌营养不良症心肌病。我们将使用经我们证明受到保护的土拨鼠从缺血再灌注损伤作为识别新的抗铁死亡途径的工具治疗和/或预防性疗法。我们将追求以下目标。目标 1：确定角色 Fe 中的 mCU 和 TRPC 在分离的心肌细胞水平和体内诱导心脏功能障碍。子目标 1-1：我们将在体外和体内证明 mCU 是否介导 Fe 吸收和 Ca 失调与 Fe 引起的心脏毒性有关。我们将确认 mito Fe 负载是由 mCU 介导的。这 Fe 治疗对线粒体功能、氧化应激和 mCU 作用的影响将得到明确。分目标 1-2：我们将证明 Fe 诱导的 TRPC 激活及其与体外（急性）和心脏功能障碍的关系体内（Fe-CM）。子目标 1-3：从子目标 1-1 和 1-2 导出的数据将用于填充计算机 E-C-M 耦合模型和结合 Fe 效应的模拟。目标 2：确定路径参与 I/R 损伤期间 Fe-CM、DMD-CM 和土拨鼠的铁死亡。分目标 2-1：我们将确定通过 mCU 进行线粒体 Fe 摄取是否在 Fe 负载铁死亡中发挥作用，并测试其他已知的铁死亡的诱导剂。我们将确定 ROS 和 TRPC 在体外和体内的作用，以及 Fe 负载和 I/R。子目标2-2：在DMD-CM心脏中，我们将确定铁死亡相关基因和蛋白质的重塑，评估生物标志物，并测试体外对铁死亡诱导剂的敏感性。我们将测试各种铁死亡途径并尝试通过抑制体内铁死亡来减轻 DMD-CM。子目标2-3：我们将获得小说深入了解土拨鼠在 I/R 期间（体外和体内）的保护机制。