Extracellular Vesicles and and HIV/cocaine associated cardiopulmonary dysfunction

项目概要： 总的来说，使用抗逆转录病毒（ART）治疗的艾滋病毒感染患者的生存期延长， 导致包括心肺功能障碍在内的非感染性并发症的发生率增加。 事实上，最近的报道表明，肺血管重构和肺动脉高压（PH）先于 在HIV感染个体中，气道破坏/肺气肿发展以及PH和COPD共存。 此外，IVDU已被发现是个体中最常见的HIV感染危险因素 诊断为HIV相关PH（HRPAH）。我们以前的研究结果一致表明， HIV感染的静脉吸毒者与HIV感染的非吸毒者或未感染者的肺血管功能障碍 静脉注射毒品因此，清楚地了解药物滥用与艾滋病病毒感染的关系，无论是单独还是在 联合引起肺血管重构的迫切需要。炎症起着至关重要的作用， 与HIV感染相关的慢性肺和心血管疾病相关的血管重塑。 我们先前在HIV感染者的肺切片中观察到显著的血管周围炎症， 猴免疫缺陷病毒（SIV）感染的猕猴暴露于滥用物质，包括增加 重塑增厚血管中的巨噬细胞数量。根据我们最近的初步调查结果，我们 假设HIV感染和可卡因介导的巨噬细胞来源的细胞外 囊泡（EV）通过调节肺动脉平滑肌细胞增殖和血管重塑， 增殖信号级联传递其货物到平滑肌细胞。这一假设将得到检验 实现四个具体目标。在使用RNA测序的第一个目标中，我们将测试EV- miRnome如何获得 影响感染艾滋病毒的巨噬细胞暴露于可卡因。在第二个目标中，我们将评估效果 这些EV对平滑肌功能的影响使用体外细胞培养系统。第三个目标将集中在 使用HIV-Tg研究EV对肺血管重构和右心功能的体内作用 最后，在第四个目标中，我们将利用来自HIV感染+/-可卡因的人类样本 滥用者将EV的变化与心肺功能障碍联系起来。这些研究具有创新性 因为据我们所知，这将是第一次尝试了解 巨噬细胞衍生的EV以及HIV-1和可卡因介导的疾病进展。拟议的研究是 重要的是，这将为理解炎症在炎症中的作用提供关键基础。 HIV-1和可卡因相关的心血管功能障碍的发展，同时在 EV作为疾病的生物标志物和/或未来预防和治疗干预的新靶点。 因此，该提案直接响应题为“艾滋病毒/艾滋病高度优先药物滥用”的PAS-16-018 研究（R 01）“将由国家药物滥用研究所（NIDA）资助。

PROJECT SUMMARY: In general, the prolonged survival of HIV-infected patients with the use of antiretroviral (ART) therapy has resulted in an increase in the incidence of non-infectious complications including cardio-pulmonary dysfunction. In fact, recent reports suggest that pulmonary vascular remodeling and pulmonary hypertension (PH) precede the airway destruction/emphysema development and that PH and COPD coexist in HIV-infected individuals. Furthermore, IVDU has been found to be the most common risk factor of HIV-infection in the individuals diagnosed with HIV related PH (HRPAH). Our previous findings consistently suggest augmentation of pulmonary vascular dysfunction in HIV infected IVDUs compared to HIV-infected non-drug users or un-infected IVDUs . Therefore, clear understanding of how the drugs of abuse and HIV-infection either alone or in combination cause pulmonary vascular remodeling is urgently needed. Inflammation plays a crucial role in vascular remodeling associated with chronic lung and cardiovascular diseases associated with HIV-infection. We earlier observed significant perivascular inflammation in the lung sections from HIV- infected humans or simian immunodeficiency virus (SIV)-infected macaques exposed to substance of abuse including increased number of macrophages in the remodeled thickened vessels. Based on our recent preliminary findings, we hypothesize that HIV-infection and cocaine mediated alterations in the macrophage derived extracellular vesicles (EVs) promote pulmonary smooth muscle hyperplasia and vascular remodeling by regulating the proliferative signaling cascades on delivery of its cargo to smooth muscle cells . This hypothesis will be tested by pursuing four specific aims. In the first aim using RNA sequencing we will test how EV- miRnome gets influenced on exposure of HIV-infected macrophages to cocaine. In the second aim, we will evaluate the effect of these EVs on smooth muscle function using in-vitro cell-culture system. Third aim will be focused on investigating the in-vivo effect of EVs on pulmonary vascular remodeling and right heart function using HIV-Tg rat model and finally in the fourth aim we will leverage on using human samples from HIV-infected +/- cocaine abusers to correlate the changes in EVs with cardio-pulmonary dysfunction. These studies are innovative because to the best of our knowledge it will be the first attempt to understand the potential link between the macrophage derived EVs and the HIV-1 and cocaine mediated disease progression. The proposed research is significant because this will provide a critical foundation to understand the role of inflammation in the development of HIV-1 and cocaine associated cardio-vascular dysfunction while identifying specific miRNAs in EVs as biomarkers of the disease and /or new targets for preventive and therapeutic interventions in future. Therefore, the proposal is directly responsive to PAS-16-018 entitled ‘HIV/AIDS High Priority Drug Abuse Research (R01)’ to be funded by National Institute on Drug Abuse (NIDA).