Quantification of HIV Reservoirs in the Gut-Associated Lymphatic System

描述（由申请人提供）： 在申请VA职业发展奖（CDA），我希望获得必要的经验，技能和知识，成为一个独立的教师调查谁使用患者驱动的研究，以更好地了解消除艾滋病毒的障碍。我的研究背景包括：1）研究原癌基因c-kit的转录控制; 2）研究病毒转录因子达特是否有助于HIV-1的潜伏感染;和3）研究肠道不同部分内的HIV水平和免疫参数。该CDA将使我能够在几个关键领域获得额外的培训，这将有助于我过渡到一个独立的翻译研究员。培训的具体领域包括先进的免疫学和病毒学，生物统计学，临床研究的设计和实施，科学合作，新的实验室技术，新的测定和赠款写作。研究环境将包括弗朗西斯科退伍军人医疗中心沿着加州大学弗朗西斯科分校（UCSF）及其附属机构，包括弗朗西斯科总医院和格莱斯顿研究所。这些机构在艾滋病毒的临床护理、培训和研究方面全国闻名。 这项研究旨在通过调查肠道是否是持续复制的场所，它是否是潜在感染的CD 4 + T细胞（或其他细胞库）的主要储库，以及如何维持这些细胞，来提高对HIV如何以及为什么持续抗逆转录病毒治疗的理解。具体目标是：1）量化艾滋病毒水平（DNA，RNA）; 2）通过测试与最近感染相关的标志物来评估回肠中正在进行的复制; 3）测量具有可诱导的、有复制能力的和/或“潜伏的”HIV的每种肠道细胞类型的比例; 4）研究肠道中潜伏的机制;和5）评估不同抗潜伏疗法从肠细胞和组织中重新激活HIV的能力。肠道组织主要通过回肠和直肠的结肠镜活检获得。此外，本研究将使用因与研究无关的临床原因接受腹部手术的患者的未使用组织。这些活组织检查和组织将用于鉴定具有最多HIV DNA和RNA的细胞群，测量肠道中的诱导型和/或复制型HIV，研究肠道中的潜伏机制，并测试不同抗潜伏疗法的效果。由此获得的知识可能最终有助于新的和改进的疗法，旨在根除艾滋病毒或与艾滋病毒终身共存。

DESCRIPTION (provided by applicant): In applying for the VA Career Development Award (CDA), I hope to acquire the experience, skills, and knowledge necessary to become an independent faculty investigator who uses patient-driven research to better understand the obstacles to eradication of HIV. My research background includes: 1) investigating the transcriptional control of the proto-oncogene c-kit; 2) investigating whether the viral transcription factor Tat contributes to latent infection with HIV-1; and 3) investigating HIV levels and immune parameters within different parts of the gut. This CDA will enable me to obtain additional training in several key areas that will facilitate my transition to an independent translational researcher. Specific areas of training include advanced immunology and virology, biostatistics, design and implementation of clinical studies, scientific collaborations, new laboratory techniques, novel assays, and grant writing. The research environment will include the San Francisco VA Medical Center along with the University of California, San Francisco (UCSF) and its affiliated institutions, including San Francisco General Hospital and the Gladstone Institute. These institutions are nationally known for clinical care, training, and research in HIV. This study seeks to improve understanding of how and why HIV persists on antiretroviral therapy by investigating whether the gut is a site of ongoing replication, whether it is a major reservoir of latently-infected CD4+ T cells (or other cellular reservoirs), and how these cells are maintained. The specific aims are: 1) to quantify levels of HIV (DNA, RNA) in different cell populations and subpopulations in both the gut and peripheral blood; 2) to assess for ongoing replication in the ileum by testing for markers that are associated with recent infection; 3) to measure the proportion of each gut cell type with inducible, replication-competent, and/or "latent" HIV; 4) to investigate mechanisms of latency in the gut; and 5) to evaluate the ability of different anti-latency therapies to reactivate HIV from gut cells and tissues. Gut tissue will be obtained primarily via colonoscopic biopsies of the ileum and rectum. In addition, the study will make use of unused tissue from patients who are undergoing abdominal surgery for clinical reasons unrelated to the study. These biopsies and tissues will be used to identify the cell populations with the most HIV DNA and RNA, to measure inducible and/or replication-competent HIV in the gut, to investigate mechanisms of latency in the gut, and to test the effect of different anti-latency therapies. The knowledge thus gained may ultimately contribute to new and improved therapies aimed at eradication or lifelong coexistence with HIV.