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: 10630874
• 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/10630874
• 关键词: Acute; Agonist; Anthracycline; Antineoplastic Agents; Blood Platelets; Blood coagulation; Cancer Patient; Cardiac; Cardiac Myocytes; Cardiotoxicity; Cell Death Induction; Cells; Chronic; Cytoprotection; Doxorubicin; Exhibits; FDA approved; Fibroblasts; G-Protein-Coupled Receptors; GTP-Binding Proteins; Generations; Goals; Heart Injuries; Heart failure; Human; In Vitro; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Modeling; Mus; Oncologist; Outcome; Oxidative Stress; PAR-1 Receptor; Pathologic; Pathology; Pathway interactions; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Prevention; Protease Inhibitor; Proteinase-Activated Receptors; Quality of life; Regimen; Role; Signal Transduction; Testing; Thrombin; Thrombin Receptor; Wild Type Mouse; activated Protein C; beta-arrestin; cancer therapy; chemotherapy; cytotoxic; design; dosage; improved outcome; in vivo; inhibitor; mouse model; novel therapeutics; pharmacologic; 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.

即使使用旨在减少心脏损伤的当前方案，Dox心脏毒性（DoxTox）仍然发生。其 预防和管理仍然是心脏病学家和肿瘤学家的主要问题， 终身剂量和损害患者的癌症结果。蛋白酶激活受体1（PAR 1）是一种G- 由心肌细胞（CM）和心脏成纤维细胞（CF）表达的蛋白偶联受体（GPCR）。它 是人类血小板上的主要凝血酶受体，也是FDA批准的药物vorapaxar的靶点。在 与人血小板相反，小鼠血小板不表达PAR 1，使其成为研究的极好模型 PAR 1的血小板非依赖性效应。PAR 1被多种蛋白酶激活。与其他GPCR一样，PAR 1 显示出有偏信号。例如，凝血酶或基质金属蛋白酶（MMP）对PAR 1的激活是一个重要因素。 细胞毒性，而活化蛋白C（APC）的活化是细胞保护性的。重要的是，我们最近 证明用凝血酶激动剂肽（AP）激活PAR 1增强Dox诱导的细胞死亡， 体外CM和CF。此外，PAR 1缺陷小鼠和野生型小鼠经vorapaxar治疗后， 急性多克斯中毒我们假设给予Dox导致凝血酶生成增加， 其他激活CM和CF上PAR 1的蛋白酶，如MMP，有助于氧化应激，导致 我们的建议有两个目标。1.确定不同PAR 1活化蛋白酶对DoxTox的影响。我们 假设凝血酶和MMPs激活的PAR 1/Gαq/Ca 2+依赖性信号增强DoxTox。在 相反，我们假设APC/PAR 1/β-arrestin 2通路将减少DoxTox。具体目标2。阐明 细胞特异性PAR 1和蛋白酶依赖性途径有助于DoxTox。我们假设PAR 1 由CM和CF表达的PAR 1/G蛋白和不同的CF特异性PAR 1/β蛋白都通过共同的PAR 1/G蛋白和不同的PAR 1/β蛋白来贡献。 急性和慢性DoxTox的抑制机制。我们假设凝血酶和基质金属蛋白酶有助于 DoxTox，而外源性APC将降低体内DoxTox。在本提案中，我们将分析PAR 1的作用 在CM上的表达与在DoxTox上的CF上的表达。此外，我们将使用药理学方法来研究 凝血酶或MMP对DoxTox的PAR 1依赖性贡献。最后，我们将测试PAR 1抑制剂或 APC可降低小鼠急性和慢性DoxTox。我们建议vorapaxar将通过阻断所有 PAR1.此外，在Dox治疗之前的APC治疗将通过增强细胞保护性来减少DoxTox。 PAR 1/β-arrestin 2信号传导。最后，偏性PAR 1抑制剂Parmodulin 2将主要减少病理性 PAR 1/G蛋白同时增强DoxTox中的保护性APC样信号传导。本提案的总体目标是 （i）了解PAR 1在DoxTox中的病理作用，（ii）开发新的治疗方法来预防蒽环类药物- 导致心脏损伤。

项目成果

Platelet in thrombo-inflammation: Unraveling new therapeutic targets.

• DOI: 10.3389/fimmu.2022.1039843
• 发表时间: 2022
• 期刊: FRONTIERS IN IMMUNOLOGY
• 影响因子: 7.3
• 作者: Sharma, Swati;Tyagi, Tarun;Antoniak, Silvio
• 通讯作者: Antoniak, Silvio

Thrombin-mediated activation of PAR1 enhances doxorubicin-induced cardiac injury in mice.

凝血酶介导的PAR1激活增强了阿霉素诱导的小鼠心脏损伤。

• DOI: 10.1182/bloodadvances.2022008637
• 发表时间: 2023-05-23
• 期刊: BLOOD ADVANCES
• 影响因子: 7.5
• 作者: Grover, Steven P.;Bharathi, Vanthana;Posma, Jens J.;Griffin, John H.;Palumbo, Joseph S.;Mackman, Nigel;Antoniak, Silvio
• 通讯作者: Antoniak, Silvio

APC-PAR1-R46 signaling limits CXCL1 expression during poly IC-induced airway inflammation in mice.

APC-PAR1-R46 信号传导在多聚 IC 诱导的小鼠气道炎症过程中限制 CXCL1 的表达。

• DOI: 10.1016/j.jtha.2023.08.018
• 发表时间: 2023
• 期刊: Journal of thrombosis and haemostasis : JTH
• 作者: Sharma,Swati;Ursery,LaurynT;Bharathi,Vanthana;Miles,StephenD;Williams,WillieA;Elzawam,AymenZ;Schmedes,ClareM;Egnatz,GrantJ;Fernandez,JoseA;Palumbo,JosephS;Griffin,JohnH;Mackman,Nigel;Antoniak,Silvio
• 通讯作者: Antoniak,Silvio