PAR1 as a Therapeutic Target in Doxorubicin-induced Cardiotoxicity

PAR1 作为阿霉素诱导的心脏毒性的治疗靶点

• 批准号: 10417140
• 负责人: Silvio Antoniak
• 金额: $ 56.37万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10417140
• 关键词: Acute; Agonist; Anthracycline; Antineoplastic Agents; Blood Platelets; Blood coagulation; Cancer Patient; Cardiac; Cardiac Myocytes; Cardiotoxicity; Cell Death; Cells; Chronic; Doxorubicin; Exhibits; FDA approved; Fibroblasts; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Generations; Goals; Heart Injuries; Heart failure; Human; In Vitro; Lead; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Modeling; Mus; Oncologist; Outcome; Oxidative Stress; PAR-1 Receptor; Pathologic; Pathology; Pathway interactions; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Pharmacology; Prevention; Protease Inhibitor; Proteinase-Activated Receptors; Quality of life; Regimen; Role; Signal Transduction; Testing; Thrombin; Thrombin Receptor; Wild Type Mouse; activated Protein C; alpha-Thrombin; cancer therapy; chemotherapy; cytotoxic; design; dosage; improved outcome; in vivo; inhibitor; mouse model; novel therapeutics; prevent; side effect; therapeutic target

Even with current regimens designed to reduce cardiac injury, Dox cardiotoxicity (DoxTox) still occurs. Its prevention and management remain a major problem for both cardiologists and oncologists, limiting the maximal lifetime dosage and compromising the patient’s cancer outcomes. Protease-activated receptor 1 (PAR1) is a G- protein coupled receptor (GPCR) that is expressed by cardiac myocytes (CM) and cardiac fibroblasts (CFs). It is the main thrombin receptor on human platelets and is the target of the FDA-approved drug vorapaxar. In contrast to human platelets, mouse platelets do not express PAR1 making them an excellent model to study platelet-independent effects of PAR1. PAR1 is activated by a variety of proteases. As with other GPCRs, PAR1 exhibits biased signaling. For instance, PAR1 activation by thrombin or matrix metalloproteinases (MMPs) is cytotoxic whereas activation by activated protein C (APC) is cytoprotective. Importantly, we recently demonstrated that activation of PAR1 with a thrombin agonist peptide (AP) enhanced Dox-induced cell death of CMs and CFs in vitro. In addition, PAR1 deficient mice and wild-type mice treated with vorapaxar were protected from acute DoxTox. We hypothesize that administration of Dox leads to increased generation of thrombin and other proteases, such as MMPs, that activate PAR1 on CMs and CFs contributes to oxidative stress, leading to DoxToxOur proposal has 2 aims. 1. Determine the effect of different PAR1 activating proteases on DoxTox. We hypothesize that PAR1/Gαq/Ca2+-dependent signaling activated by thrombin and MMPs enhances DoxTox. In contrast, we hypothesize that the APC/PAR1/β-arrestin2 pathway will reduce DoxTox. Specific Aim 2. Elucidate the cell-specific PAR1- and protease-dependent pathways contributing to DoxTox. We hypothesize that PAR1 expressed by CMs and CFs both contribute via common PAR1/G-protein and distinct CF-specific PAR1/β- arrestin1 mechanisms to acute and chronic DoxTox. We hypothesize that thrombin and MMPs contribute to DoxTox whereas exogenous APC will reduce DoxTox in vivo. In this proposal, we will analyze the role of PAR1 expressed on CMs versus CFs on DoxTox. In addition, we will use a pharmacologic approach to investigate the PAR1-dependent contribution of thrombin or MMPs to DoxTox. Finally, we will test whether PAR1 inhibitors or APC reduce acute and chronic DoxTox in mice. We propose that vorapaxar will reduce DoxTox by blocking all PAR1. In addition, APC treatment prior to Dox treatment will reduce DoxTox by enhancing cytoprotective PAR1/β-arrestin2 signaling. Finally, the biased PAR1 inhibitor Parmodulin 2 will reduce primarily pathologic PAR1/G-protein while enhancing protective APC-like signaling in DoxTox. The overall goals of this proposal are to (i) understand the pathologic roles of PAR1 in DoxTox and (ii) develop new therapies to prevent anthracycline- induced cardiac injury.

即使采用目前旨在减少心脏损伤的治疗方案，Dox 心脏毒性 (DoxTox) 仍然会发生。 预防和管理仍然是心脏病学家和肿瘤学家的一个主要问题，限制了最大的 终生剂量并损害患者的癌症结果。蛋白酶激活受体 1 (PAR1) 是一种 G-。 心肌细胞 (CM) 和心脏成纤维细胞 (CF) 表达的蛋白偶联受体 (GPCR)。 是人类血小板上的主要凝血酶受体，也是 FDA 批准的药物 vorapaxar In 的靶点。 与人类血小板相比，小鼠血小板不表达 PAR1，这使其成为研究的绝佳模型 PAR1 的血小板依赖性作用与其他 GPCR 一样，由多种蛋白酶激活。 例如，凝血酶或基质金属蛋白酶 (MMP) 激活 PAR1 是有偏差的。 重要的是，我们最近发现，活化蛋白 C (APC) 的激活具有细胞毒性。 证明用凝血酶激动剂肽（AP）激活 PAR1 可增强 Dox 诱导的细胞死亡 此外，用沃拉帕沙治疗的 PAR1 缺陷小鼠和野生型小鼠受到保护。 我们勇敢地说，服用 Dox 会导致凝血酶和凝血酶的产生增加。 其他蛋白酶，例如 MMP，可激活 CM 和 CF 上的 PAR1，从而导致氧化应激，从而导致 DoxTox 我们的提案有 2 个目标 1. 确定不同 PAR1 激活蛋白酶对 DoxTox 的影响。 研究表明，凝血酶和 MMP 激活的 PAR1/Gαq/Ca2+ 依赖性信号传导可增强 DoxTox In。 相比之下，我们发现 APC/PAR1/β-arrestin2 途径会减少 DoxTox。具体目标 2。阐明。 导致 DoxTox 的细胞特异性 PAR1 和蛋白酶依赖性途径 由 CM 和 CF 表达的蛋白均通过共同的 PAR1/G 蛋白和独特的 CF 特异性 PAR1/β- 贡献 我们发现凝血酶和 MMP 有助于急性和慢性 DoxTox 的抑制蛋白 1 机制。 DoxTox 而外源性 APC 会减少体内的 DoxTox 在本提案中，我们将分析 PAR1 的作用。 此外，我们将使用药理学方法来研究 CM 与 DoxTox 上的 CF 的差异。 凝血酶或 MMP 对 DoxTox 的 PAR1 依赖性贡献 最后，我们将测试 PAR1 抑制剂或 DoxTox。 APC 减少小鼠的急性和慢性 DoxTox 我们建议 vorapaxar 将通过阻断所有药物来减少 DoxTox。 此外，在 Dox 治疗之前进行 PAR1 治疗将通过增强细胞保护作用来减少 DoxTox。 最后，偏向的 PAR1 抑制剂 Parmodulin 2 将主要减少病理性。 PAR1/G 蛋白同时增强 DoxTox 中的保护性 APC 样信号传导 该提案的总体目标是 (i) 了解 PAR1 在 DoxTox 中的病理作用，以及 (ii) 开发新疗法来预防蒽环类药物- 诱发心脏损伤。