The Role of Mitochondrial DNA in Innate Immune Activation after Sudden CardiacArrest

线粒体 DNA 在心脏骤停后先天免疫激活中的作用

• 批准号: 10656384
• 负责人: Cody Rutledge
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656384
• 关键词: Admission activity; Adult; Affect; American; Anti-Inflammatory Agents; Bacterial DNA; Cardiac; Cardiac Myocytes; Cardiac Volume; Cardiogenic Shock; Cardiovascular Diseases; Cell Death; Cell model; Cells; Characteristics; Chemicals; Clinical; Clinical Research; Cyclic GMP; Cytokine Signaling; Cytoplasm; Data; Development; Disease model; Flow Cytometry; Functional disorder; Future; Gene Proteins; Genes; Goals; Grant; Heart; Heart Arrest; Heart Diseases; High Prevalence; Hospital Administration; Hospitalization; Hospitals; Human; Hypoxia; Immune response; Immune signaling; In Vitro; Induced Heart Arrest; Inflammation; Inflammatory; Inflammatory Response; Injury; Innate Immune Response; Innate Immune System; Intensive Care; Ischemia; Knock-out; Knockout Mice; LoxP-flanked allele; Mediating; Mentorship; Microscopy; Mitochondria; Mitochondrial DNA; Molecular; Multiple Organ Failure; Mus; Myocardial dysfunction; Myocarditis; Myocardium; Nuclear; Operative Surgical Procedures; Organ; Outcome; Pathologic; Patients; Pattern; Peripheral; Permeability; Pharmaceutical Preparations; Physicians; Recovery; Reperfusion Injury; Research; Resuscitation; Role; Scientist; Septic Shock; Serum; Signal Pathway; Signal Transduction; Stimulator of Interferon Genes; Structure; Survivors; Syndrome; System; Testing; Therapeutic Intervention; Time; Tissues; Training; Transgenic Mice; Translational trial; United States; United States Department of Veterans Affairs; Universities; Veterans; Visualization; Work; career; career development; circulating DNA; clinically relevant; cytokine; experimental study; extracellular; factor A; genetic manipulation; heart function; immune activation; improved; improved outcome; in vivo Model; inflammatory marker; military veteran; mortality; mouse model; mtTF1 transcription factor; novel; overexpression; pharmacologic; preservation; prevent; research and development; response; sudden cardiac death; survival outcome; therapeutic target; transcription factor; transcriptome sequencing

ABSTRACT Sudden cardiac arrest is highly prevalent among hospitalized veterans and results in overwhelming mortality. Unfortunately, there are no pharmacologic therapies that have been shown to reliably increase survival after sudden cardiac arrest. Survivors of sudden cardiac arrest typically have systemic organ damage requiring intensive care in the hospital. The majority of these patients have reduced cardiac function and one quarter of these patients die from cardiogenic shock. Cardiac inflammation is thought to contribute to this dysfunction and is likely driven by damage associated molecular patterns (DAMPs). One prominent DAMP is mitochondrial DNA (mtDNA). Preliminary work in our lab has shown that strategies aimed at preserving mtDNA integrity, including overexpression of mitochondrial transcription factor A (TFAM), are protective to cardiac function in a mouse model of sudden cardiac arrest. TFAM is a nuclear gene that regulates mtDNA expression, packaging, and copy number and is known to be protective in a number of heart disease models. My overarching hypothesis is that ischemia-reperfusion injury from cardiac arrest results in mtDNA release, which triggers an inflammatory response in the heart, and that this response is dependent upon cGAS/STING signaling. To explore this hypothesis, I will pursue three specific aims. In Aim 1, I will explore the mechanism by which mtDNA is released using microscopy in vitro and confirm these changes in an in vivo model of sudden cardiac arrest. In Aim 2, I will use transgenic mouse models to manipulate TFAM to affect mtDNA release and also knock-out STING signaling to explore the relationship between mtDNA release and inflammation after sudden cardiac arrest. In Aim 3, I will characterize the inflammatory response in the heart after sudden cardiac arrest and manipulate cGAS/STING signaling to evaluate cGAS/STING mediated inflammatory changes. Together, these aims will evaluate the role of mtDNA release following sudden cardiac arrest and its role in innate immune signaling, which may guide future therapies for human studies. This work will support my goal of transitioning into an independent research career as a physician-scientist studying mitochondrial changes in cardiovascular disease. This grant will support continued research and career development at both the Veterans Administration Hospital in Pittsburgh and the University of Pittsburgh, including coursework, career mentorship, and scientific training aimed at transitioning to independence as a physician scientist in the Veterans Administration system.

抽象的 在住院的退伍军人中，突然的心脏骤停高度普遍，导致压倒性的死亡率。 不幸的是，没有药理学疗法已证明可以可靠地增加生存率 突然心脏骤停。突然心脏骤停的幸存者通常会有系统性的器官损害 医院重症监护。这些患者的大多数患者的心脏功能降低，四分之一 这些患者死于心源性休克。心脏炎症被认为会导致这种功能障碍和 可能是由损害相关的分子模式（湿）驱动的。一种突出的潮湿是线粒体 DNA（mtDNA）。我们实验室中的初步工作表明，旨在维护mtDNA完整性的策略， 包括线粒体转录因子A（TFAM）的过表达对A中的心脏功能具有保护作用 小鼠心脏骤停模型。 TFAM是一种调节mtDNA表达，包装， 和拷贝数，已知在许多心脏病模型中具有保护性。 我的总体假设是，在mtDNA释放中，心脏骤停的缺血再灌注损伤， 这会触发心脏的炎症反应，并且这种反应取决于CGA/STING 信号。为了探讨这一假设，我将追求三个具体目标。在AIM 1中，我将探索机制 使用显微镜在体外释放mtDNA，并在体内模型中确认这些变化 突然心脏骤停。在AIM 2中，我将使用转基因小鼠模型来操纵TFAM以影响mtDNA 释放以及敲除刺激信号传导，以探索mtDNA释放与 突然心脏骤停后炎症。在AIM 3中，我将表征心脏中的炎症反应 突然心脏骤停和操纵CGA/sting信号传导以评估CGA/STING介导 炎症变化。 这些目标共同评估心脏骤停后mtDNA释放的作用及其在 先天免疫信号传导，这可能指导未来的人类研究疗法。这项工作将支持我的目标 作为医师科学家研究线粒体变化的医师科学家，过渡到独立研究职业 心血管疾病。该赠款将支持持续的研究和职业发展 匹兹堡的退伍军人管理医院和匹兹堡大学，包括课程，职业 指导和科学培训旨在转变为独立的医师科学家 退伍军人管理系统。