Potential Role of Extracellular Vesicles for the Development of HIV Comorbidities

项目摘要 因有效的抗逆转录病毒治疗(ART)病毒载量低于目标水平的艾滋病毒感染者 75%以上的慢性艾滋病毒携带者患心肺疾病的风险继续增加 表现为临床症状的疾病。申请者实验室最近的研究提供了证据 HIV蛋白，特别是HIV-Nef被有效地保留在HIV患者的血浆和肺液中 联合抗逆转录病毒疗法(ART)。基于这项先前的工作，我们计划在体外阐明临床前 并在体内模拟血管内皮损伤和过早衰老的机制。我们的主要假设是 细胞内的HIV蛋白与HIV-Nef一起从细胞中释放出来，并通过细胞外的小泡运输 (EV)引起心肺变化，导致合并症。在目标1中，我们将研究HIV-蛋白质 细胞外小泡及其与特定货物的关系，主要集中在表面和内小泡上 定位。对特定EV相关HIV蛋白表面标记的检测将允许未来 治疗和诊断应用，包括基于抗体的靶向技术。在目标2中，我们将 分析EV相关HIV蛋白在将HIV-EV相关货物运送到全身的过程中所起的作用 增加炎症和细胞衰老。具体地说，我们将使用多个抗体小组来确定细胞 “归宿”电动汽车的身份和位置。在目标3中，我们将重点介绍交付的艾滋病毒-EV的病理 相关货物。作为原则的证明，我们将测试具体的干预策略，包括ADAM17 通过EV传递HIV-蛋白质的临床前小鼠模型中的抑制物和老年溶解剂。

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.