Potential Role of Extracellular Vesicles for the Development of HIV Comorbidities

项目摘要 由于有效的抗逆转录病毒疗法（ART），病毒载量低于目标水平的艾滋病毒感染者 继续增加患心肺疾病的风险，超过75%的慢性艾滋病毒感染者 有临床表现的疾病。申请人实验室最近的研究提供了证据， HIV蛋白，特别是HIV-Nef，在有效治疗后保留在HIV患者的血浆和肺液中， 联合抗逆转录病毒疗法（ART）。基于这一先前的工作，我们计划在临床前体外阐明 并在体内模拟内皮损伤和过早衰老的机制。我们的主要假设是 细胞内HIV蛋白与HIV-Nef一起从细胞中释放出来，并通过细胞外囊泡 (EV)导致心肺功能改变导致合并症。在目标1中，我们将研究 细胞外囊泡及其与特定货物的关联，重点是表面和囊泡内 导向检测特定EV相关HIV蛋白的表面标志物，将允许未来 治疗和诊断应用，包括基于抗体的靶向技术。在目标2中，我们 分析EV相关的HIV蛋白在整个身体中递送HIV-EV相关货物的作用， 增加炎症和细胞衰老。具体来说，我们将使用多抗体面板来确定细胞 身份和位置的“归巢”电动车。在目标3中，我们将重点关注所递送的HIV-EV的病理学 相关货物。作为原则的证明，我们将测试特定的干预策略，包括ADAM 17 抑制剂和衰老清除剂用于通过EV递送HIV蛋白的临床前小鼠模型。

Project Summary HIV-infected people whose viral load is below target levels due to effective anti-retroviral therapy (ART) continue to be at increased risk for cardio-pulmonary disease with over 75% of patients with chronic HIV disease showing clinical manifestations. Recent studies in the laboratories of the applicants provided evidence that HIV proteins and in particular HIV-Nef is retained in plasma and lung fluids of HIV patients on effective combined anti-retroviral therapy (ART). Based on this previous work we plan to elucidate in preclinical in vitro and in vivo models the mechanism of endothelia damage and premature aging. Our main hypothesis is that intracellular HIV proteins are released from cells together with HIV-Nef and travel through extracellular vesicles (EV) to cause cardiopulmonary changes leading to comorbidities. In aim 1 we will study HIV-proteins in extracellular vesicles and their association with specific cargo with focus on surface- and intra-vesicular orientation. The detection of surface markers for specific EV-associated HIV proteins, will allow for future therapeutic and diagnostic applications including antibody-based targeting techniques. In aim 2, we will analyze the role EV-associated HIV proteins in delivering HIV-EV-associated cargo throughout the body to increase inflammation and cell senescence. Specifically, we will use multi-antibody panels to determine cell identity and location of the “homed” EV. In aim 3, we will focus on the pathology of the delivered HIV-EV associated cargo. As a proof of principal we will test specific intervention strategies including ADAM17 inhibitors and senolytic agents in preclinical mouse models for HIV-protein delivery through EV.