Quantification of HIV Reservoirs in the Gut-Associated Lymphatic System

描述(由申请人提供)： 在申请退伍军人管理局职业发展奖(CDA)时，我希望获得必要的经验、技能和知识，成为一名独立的教员调查员，利用患者驱动的研究更好地了解根除艾滋病毒的障碍。我的研究背景包括：1)调查原癌基因c-kit的转录调控；2)调查病毒转录因子Tat是否与HIV-1潜伏感染有关；3)调查肠道不同部位的HIV水平和免疫参数。这一CDA将使我能够在几个关键领域获得额外的培训，这将促进我向独立翻译研究员的过渡。培训的具体领域包括高级免疫学和病毒学、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、新的分析方法和拨款申请。研究环境将包括旧金山退伍军人医学中心以及加州大学旧金山分校(UCSF)及其附属机构，包括旧金山总医院和格莱斯顿研究所。这些机构以艾滋病毒的临床护理、培训和研究而闻名全国。这项研究试图通过调查肠道是否是持续复制的部位，它是否是潜伏感染的CD4+T细胞(或其他细胞储存库)的主要储备库，以及这些细胞是如何维持的，来提高对艾滋病毒如何以及为什么坚持接受抗逆转录病毒治疗的理解。其具体目标是：1)量化肠道和外周血中不同细胞群和亚群中的艾滋病毒(DNA、RNA)水平；2)通过检测与最近感染相关的标记物，评估回肠正在进行的复制；3)测量每种肠道细胞类型中可诱导、有复制能力和/或“潜伏”艾滋病毒的比例；4)研究肠道潜伏期的机制；以及5)评估不同抗潜伏期疗法从肠道细胞和组织重新激活艾滋病毒的能力。肠道组织将主要通过回肠和直肠的结肠镜活检获得。此外，这项研究将利用因与研究无关的临床原因而接受腹部手术的患者的未使用组织。这些活检和组织将用于识别具有最多HIV DNA和RNA的细胞群，测量肠道中可诱导和/或复制能力的HIV，研究肠道潜伏期的机制，并测试不同抗潜伏期疗法的效果。由此获得的知识最终可能有助于旨在根除或终身与艾滋病毒共存的新的和改进的治疗方法。 公共卫生相关性： 目前，超过22,000名美国艾滋病毒携带者通过退伍军人管理局接受治疗，使退伍军人管理局成为美国最大的艾滋病毒+患者医疗保健提供者。尽管抗病毒药物彻底改变了艾滋病毒携带者的生活，但这些疗法昂贵，需要终身给药和监测，造成副作用和毒性，无法防止艾滋病毒造成的持续损害(平均损失11年生命)，而且很难提供给所有艾滋病毒携带者。因此，探索新的、不同的艾滋病毒治疗方法似乎势在必行，特别是那些可能导致根除的治疗方法。这项研究旨在进一步定义和量化已知的最大艾滋病毒宿主(肠道内)，探索可能使艾滋病毒持续接受抗病毒治疗的机制(包括肠道内的潜伏感染和持续复制)，并评估旨在克服肠道或血液中潜伏感染的治疗方法。通过这样做，它有可能带来新的和改进的艾滋病毒治疗方法，这将对包括美国退伍军人在内的所有艾滋病毒+患者产生巨大影响。

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. PUBLIC HEALTH RELEVANCE: Over 22,000 U.S. veterans with HIV currently receive care through VA facilities, making the VA the largest provider of medical care to HIV+ patients in the U.S. Though antiviral medicines have revolutionized the lives of those with HIV, these therapies are expensive, require lifelong administration and monitoring, cause side effects and toxicities, do not prevent ongoing damage from HIV (average loss of 11 years of life), and are difficult to provide to everybody with HIV. Thus, it seems imperative to explore new and different therapies for HIV, especially those that can possibly lead to eradication. This study seeks to further define and quantify the largest known reservoir of HIV (in the gut), to explore mechanisms that may allow HIV to persist on antiviral therapy (including latent infection and ongoing replication in the gut), and to evaluate therapies aimed at overcoming latent infection in the gut or blood. In doing so, it has the potential to lead to new and improved therapies for HIV, which would have an enormous impact on all HIV+ patients, including U.S. veterans.