Cardiovascular Risk of Antiretroviral Therapy Drugs in HIV

项目总结 尽管抗逆转录病毒治疗药物(ART)阻止了艾滋病毒的传播并延长了患者的预期寿命 艾滋病毒携带者(PWH)，这一人群的猝死率比未感染艾滋病毒的人高2-4倍。 尸检显示，这些HIV患者中有一半患有心脏纤维化，这可能是猝死的原因之一。 蛋白水解酶抑制物类ART(PI-ART)与PWH的心血管风险有关，ART似乎是合理的 可通过诱导心脏纤维化而加剧风险。我们的长期目标是确定机制和 ART诱导的心脏纤维化对PWH的影响，并探讨预防策略。转化生长因子β1 转化生长因子β1是一种强促肝纤维化的细胞因子，其分泌的转化生长因子β1是其他细胞的100倍。 血浆转化生长因子β-1是导致器官纤维化的主要来源。血浆转化生长因子β-1水平升高与心肌纤维化 在艾滋病患者中观察到，但ART是否进一步增加了重症肝炎患者的血浆转化生长因子β1和心脏纤维化 不清楚。在试点研究中，我们观察到较新的ART方案，包括PI增强剂量的利托那韦(RTV)和 替诺福韦激活血小板释放转化生长因子β-1，可被MRP4的特异性抑制剂头孢富林-1阻断， 一种膜转运蛋白，在HIV患者的血小板中高表达。注射PI增强剂量的RTV 在转基因TG26艾滋病毒小鼠(表现出多种艾滋病毒相关共病)中，心脏纤维化和 与表达αSMA的CD206细胞在心脏积聚有关的舒张期功能障碍 可能是巨噬细胞。这些结果导致了我们的中心假设，即ART可能会激活血小板释放 转化生长因子β-1通过MRP4刺激巨噬细胞间充质转化，诱导心肌纤维化。 该应用的目的是确定不同类别的ART诱导血小板的机制。 转化生长因子β1释放并鉴定转化生长因子β1信号转导的细胞类型，导致心肌纤维化。以下是具体的 AIMS将致力于实现这一目标：1)筛选不同类别的艺术，单独或组合，用于 诱导转化生长因子β-1释放血小板并确定这一过程的机制；2)确定 当代ART介导的转化生长因子β1通过MRP4的释放在体内诱导心肌纤维化；3)确定 巨噬细胞内转化生长因子β-1信号转导导致间质转化和心肌纤维化。我们的研究将澄清 ART诱导血小板释放转化生长因子β-1的机制及转化生长因子-β-1信号转导的细胞类型 到心脏纤维化。我们将使用创新的技术来评估血小板的激活，检测血浆转化生长因子β1 水平，并在两种HIV小鼠模型中评估细胞信号和心脏纤维化。此外，我们的研究 将探索通过抗转化生长因子β1和抗ART诱导的艾滋病毒小鼠心脏纤维化的翻译潜力 抗纤维化药物，如加鲁尼司替布或吡非尼酮。我们的研究也可能阐明转化生长因子β1是否可以 可作为PWH潜在脏器纤维化的生物标志物。它还可能为更好的机械化奠定基础 了解并提出新的策略，以预防重症肝炎和其他感染性疾病中的纤维化。

PROJECT SUMMARY Although antiretroviral therapy drugs (ART) have prevented HIV propagation and increased life expectancy of people with HIV (PWH), the rate of sudden death in this population is 2-4-times higher than people without HIV. Autopsies have revealed cardiac fibrosis in half of this HIV patient population, a likely etiology for sudden death. The protease inhibitor class ART (PI-ART) is linked to cardiovascular risk in PWH, and it is plausible that ART can exacerbate the risk by inducing cardiac fibrosis. Our long-term goals are to determine the mechanism and the impact of ART-induced cardiac fibrosis in PWH, and to explore preventive strategies. Transforming growth factor β1 (TGFβ1) is a strong profibrotic cytokine and platelets contain ~100 times more TGFβ1 than other cells and are a major source of plasma TGFβ1 contributing to organ fibrosis. Higher plasma TGFβ1 levels and cardiac fibrosis are observed in HIV+ individuals, but whether ART further increases plasma TGFβ1 and cardiac fibrosis in PWH is not clear. In pilot studies, we observed that newer ART regimens, including PI-boosted dose of ritonavir (RTV) and tenofovir, activated platelets to release TGFβ1, which can be blocked by Ceefourin-1, a specific inhibitor of MRP4, a membrane transporter highly expressed in platelets from HIV patients. Injection of a PI-boosted dose of RTV in transgenic Tg26 HIV mice (which exhibit multiple HIV-associated comorbidities) increased cardiac fibrosis and diastolic dysfunction associated with the accumulation of CD206+ cells expressing αSMA in the heart, presumably macrophages. These results led to our central hypothesis that ART may activate platelets to release TGFβ1 via MRP4, which stimulates macrophages to undergo mesenchymal transition, inducing cardiac fibrosis. The objective of this application is to determine the mechanism by which different classes of ART induce platelet TGFβ1 release and identify the cell types to which TGFβ1 signals, leading to cardiac fibrosis. The following Specific Aims will address the objective: 1) Screen a panel of different classes of ART, alone and in combination, for induction of platelet release of TGFβ1 and identify the mechanisms of this process; 2) Determine whether contemporary ART-mediated TGFβ1 release via MRP4 induces cardiac fibrosis in vivo; 3) Determine whether TGFβ1 signaling in macrophages leads to mesenchymal transition and cardiac fibrosis. Our studies will clarify the mechanism of ART-induced TGFβ1 release from platelets and the cell types on which TGFβ1 signals, leading to cardiac fibrosis. We will use innovative techniques to evaluate platelet activation, measure plasma TGFβ1 levels, and assess cellular signaling and cardiac fibrosis in two murine models of HIV. Furthermore, our research will explore the translational potential for mitigating ART-induced cardiac fibrosis in HIV mice with anti-TGFβ1 and anti-fibrotic agents, such as Galunisertib or Pirfenidone. Our studies may also elucidate whether TGFβ1 could be a potential biomarker of underlying organ fibrosis in PWH. It may also lay the foundation for better mechanistic understanding and novel strategies for preventing comorbidities in PWH and fibrosis in other infectious diseases.