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700万人，造成17000人死亡，每年造成约80亿美元的医疗费用和损失 生产力。感染者会出现氧化和炎症应激，脑室纤维化和扩张， 并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 Chagaic病理学，并提供一种创新的潜在疗法。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 心肌细胞线粒体活性氧(ROS)生成的效率和增加 查吉斯之心。此外，ROS诱导的细胞碎片吞噬以及PARP1依赖的代谢 巨噬细胞开关信号是促炎症巨噬细胞活化和增殖的信号。我们将聘用 创新的、基于荧光的分析方法，测量同一样本中的多个功能反应以进行测试 我们的假设有两个具体目的。 在目标1中，我们的目标是证明PARP1的激活增加了临床心脏的风险 感染患者中的疾病，解剖PARP1如何干扰mtDNA复制体，并随着病情的加重 心脏病，以及靶向向线粒体输送PARP1抑制剂保护线粒体的试验 恰加斯病患者的健康和左心功能。 在目标2中，我们目标是测试由于ROS/PARP1诱导产生的细胞外小泡(EV 细胞损伤带有慢性恰加斯病的免疫特征。我们将展示电动汽车，在 疾病阶段特异性方式，激活巨噬细胞的细胞内天然免疫受体，以及 巨噬细胞表达PARP1为糖酵解开关和促炎性m-φ提供代谢信号 激活。重要的是，我们将测试控制PARP1的激活对组织沉默是有益的- 萎缩性贫血患者巨噬细胞的破坏性炎性表型。 我们认为，创新之处在于展示了DNA修复蛋白如何干扰 线粒体功能和加剧炎症。我们将提供机械性的洞察力，了解这些 过程是相互联系的，并提供了一种新的治疗方法，以保持代谢稳态和左心功能。 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.