Oxidative Response Networks in Chagasic Cardiomyopathy

摘要 由克氏锥虫引起的恰加斯病是第三大热带疾病负担。光盘 影响超过 700 万人，导致超过 17000 人死亡，每年造成约 80 亿美元的医疗保健费用和损失 生产力。感染个体呈现氧化和炎症应激、心室纤维化和扩张， 最终发展为充血性心力衰竭。 在这个项目中，我们建议研究聚（ADP-核糖）聚合酶 1 (PARP1) 在 恰加斯病理学并提供创新的潜在疗法。简而言之，我们认为 PARP1 与 线粒体 DNA 聚合酶 G (POLG) 影响线粒体 DNA 完整性，导致呼吸链下降 心肌细胞中线粒体活性氧 (ROS) 产生的效率和增加 恰加斯心。此外，ROS 诱导的细胞碎片的吞噬作用以及 PARP1 依赖性代谢 巨噬细胞的开关信号促炎巨噬细胞的激活和增殖。我们将聘用 创新的、基于荧光的检测方法，可测量同一样品中的多种功能反应以进行测试 我们的假设有两个具体目标。 在目标 1 中，我们的目标是证明 PARP1 激活会增加临床心脏病的风险 受感染患者的疾病，剖析 PARP1 如何干扰 mtDNA 复制体，并随着疾病严重程度的增加 心脏病，并测试将 PARP1 抑制剂靶向递送至线粒体可保护线粒体 恰加斯病的健康和左心室功能。 在目标 2 中，我们的目标是测试 ROS/PARP1 诱导产生的细胞外囊泡 (EV) 细胞损伤带有慢性恰加斯病的免疫特征。我们将证明电动汽车 疾病阶段特异性方式，参与巨噬细胞的细胞内先天免疫受体，以及 PARP1 的巨噬细胞表达为糖酵解开关和促炎性 mφ 提供代谢信号 激活。重要的是，我们将测试控制 PARP1 激活是否有利于沉默组织- 恰加斯病患者巨噬细胞的破坏性炎症表型。 我们相信这项创新在于展示 DNA 修复蛋白如何干扰 线粒体功能并加剧炎症。我们将提供有关这些如何发生的机制见解 这些过程是相互关联的，并为维持代谢稳态和左心室功能提供了一种新的疗法 恰加斯病病例。

ABSTRACT Chagas disease, caused by Trypanosoma cruzi, represents the third greatest tropical disease burden. CD affects >7 million people, causes >17000 deaths, and costs ~$8.0 billion per year in health care costs and lost productivity. Infected individuals present oxidative and inflammatory stress, ventricular fibrosis and dilatation, and eventually develop congestive heart failure. In this project, we propose to examine a novel role of poly (ADP-ribose) polymerase 1 (PARP1) in chagasic pathology and offer an innovative potential therapy. Briefly, we believe that PARP1 cross-talk with mitochondrial DNA polymerase G (POLG) effects the mtDNA integrity, leading to a decline in respiratory chain efficiency and increase in mitochondrial reactive oxygen species (ROS) production in cardiomyocytes and chagasic heart. Moreover, phagocytosis of ROS-induced cell debris along with PARP1-dependent metabolic switch in macrophages signals activation and proliferation of proinflammatory macrophages. We will employ innovative, fluorescence-based, assays that measure multiple functional responses in the same sample to test our hypothesis in two specific aims. In aim 1, our objectives are to demonstrate that PARP1 activation increases the risk of clinical heart disease in infected patients, dissect how PARP1 interferes with mtDNA replisome with increasing severity of heart disease, and test that targeted delivery of PARP1 inhibitors to mitochondria preserves mitochondrial health and LV function in Chagas disease. In aim 2, our objectives are to test that extracellular vesicles (EV) produced due to ROS/PARP1-induced cellular injury carry the immune signature of chronic Chagas disease. We will demonstrate that EVs, in a disease stage-specific manner, engage intracellular innate immune receptors of macrophages, and macrophage expression of PARP1 provides metabolic signal for glycolytic switch and proinflammatory mφ activation. Importantly, we will test that controlling PARP1 activation is beneficial in silencing the tissue- destructive, inflammatory phenotype of chagasic patients’ macrophages. We believe the innovation lies in the idea of demonstrating how a DNA repair protein can disturb mitochondrial function and intensify inflammation. We will provide mechanistic insights into how these processes are linked and offer a novel therapy for preserving metabolic homeostasis and LV function in Chagas disease cases.