Cardiovascular Risk of Antiretroviral Therapy Drugs in HIV

项目概要 尽管抗逆转录病毒治疗药物（ART）已经阻止了艾滋病毒的传播并延长了人们的预期寿命 HIV 感染者 (PWH) 的猝死率比未感染 HIV 的人高 2-4 倍。 尸检显示，一半的艾滋病毒患者患有心脏纤维化，这可能是猝死的病因。 蛋白酶抑制剂类 ART (PI-ART) 与孕妇心血管风险相关，ART 似乎是合理的 可能会诱发心脏纤维化，从而加剧风险。我们的长期目标是确定机制和 ART 诱发的心脏纤维化对 PWH 的影响，并探索预防策略。转化生长因子β1 (TGFβ1) 是一种强效促纤维化细胞因子，血小板中的 TGFβ1 含量比其他细胞多约 100 倍，是一种 血浆 TGFβ1 导致器官纤维化的主要来源。血浆 TGFβ1 水平升高与心脏纤维化有关 在 HIV 个体中观察到，但 ART 是否会进一步增加 PWH 中的血浆 TGFβ1 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 TGFβ1，可被 Ceefourin-1（MRP4 的特异性抑制剂）阻断， 一种在艾滋病毒患者血小板中高表达的膜转运蛋白。注射 PI 增强剂量的 RTV 在转基因 Tg26 HIV 小鼠（表现出多种 HIV 相关合并症）中，心脏纤维化增加 与心脏中表达 αSMA 的 CD206 细胞积聚相关的舒张功能障碍， 推测是巨噬细胞。这些结果引出了我们的中心假设：ART 可能会激活血小板释放 TGFβ1 通过 MRP4 刺激巨噬细胞进行间质转化，诱导心脏纤维化。 本申请的目的是确定不同类别的 ART 诱导血小板减少的机制 TGFβ1 释放并识别 TGFβ1 信号传导的细胞类型，从而导致心脏纤维化。具体如下 目的将解决以下目标： 1) 筛选不同类别 ART 的小组，单独或组合，以 诱导血小板释放 TGFβ1 并确定该过程的机制； 2）判断是否 当代 ART 介导的 TGFβ1 通过 MRP4 释放可诱导体内心脏纤维化； 3）判断是否 巨噬细胞中的 TGFβ1 信号传导导致间质转化和心脏纤维化。我们的研究将澄清 ART 诱导血小板释放 TGFβ1 的机制以及 TGFβ1 信号传导的细胞类型，导致 到心脏纤维化。我们将使用创新技术来评估血小板活化、测量血浆 TGFβ1 水平，并评估两种 HIV 小鼠模型中的细胞信号传导和心脏纤维化。此外，我们的研究 将探索使用抗 TGFβ1 药物减轻 ART 诱导的 HIV 小鼠心脏纤维化的转化潜力 抗纤维化药物，例如 Galunisertib 或吡非尼酮。我们的研究还可能阐明 TGFβ1 是否可以 是 PWH 潜在器官纤维化的潜在生物标志物。它还可能为更好的机制奠定基础 预防感染者合并症和其他传染病纤维化的理解和新策略。

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.