Quantification of HIV-1 Reservoirs in the Gut

描述（申请人提供）：我的研究背景包括：1)研究基因c-kit的转录控制；2)研究病毒转录因子Tat是否与HIV-1潜伏感染有关；3)研究肠道不同部位的HIV水平和免疫参数。在申请美国国立卫生研究院职业发展奖（CDA）时，我希望获得必要的经验、技能和知识，成为一名独立的教师研究者，利用患者驱动的研究来更好地了解根除艾滋病毒的障碍。这个CDA将使我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。研究环境将包括加州大学旧金山分校和三个附属机构：旧金山总医院、旧金山退伍军人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。本研究旨在通过调查肠道中是否存在持续复制，肠道是否是潜伏感染CD4+ T细胞的主要储存库，以及新的抗逆转录病毒药物或抗潜伏治疗是否可以影响肠道中正在进行的复制和/或潜伏感染，从而提高对艾滋病毒在抗逆转录病毒治疗（ART）中持续存在的方式和原因的理解。具体目的是：1)比较肠道内不同区域和细胞类型的HIV水平和免疫参数；2)通过测量治疗强化对HIV、免疫标记物和异常肠道屏障功能标记物水平的影响来评估在肠道中的持续复制；3)评估肠道潜伏库的大小，研究肠道潜伏的机制，评估肠道细胞和组织对抗潜伏治疗的反应。研究第一部分是一项前瞻性、纵向的试点研究，参与者将接受基线上、下内窥镜检查，并从4个区域进行活检，随后进行12周的强化治疗（使用雷替重力韦+/-第二种药物）和重复内窥镜检查。该研究的第二部分将包括内窥镜检查，从一个肠道部位（2A部分）进行更密集的采样，以及因其他原因接受腹部手术的HIV+患者的未使用组织（2B）。这些活组织检查和组织将用于鉴定含有最多HIV DNA和RNA的细胞群，建立培养系统以测量肠道中可诱导和/或可复制的HIV，研究肠道中潜伏的机制，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治疗方法，旨在根除或终生与艾滋病毒共存。虽然抗病毒药物彻底改变了艾滋病毒感染者的生活，但这些疗法价格昂贵，需要终身给药和监测，有副作用和毒性的风险，不能根除艾滋病毒，不能完全扭转艾滋病毒的免疫缺陷，而且很难向所有艾滋病毒感染者提供。因此，探索新的和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或提高终身艾滋病毒感染耐受性的治疗方法。本研究旨在通过研究肠道中是否存在持续的病毒复制，肠道是否是潜伏感染细胞的主要储存库，以及新疗法是否可以影响肠道中的持续复制和/或潜伏感染，来提高对HIV持续治疗的方式和原因的理解。

DESCRIPTION (provided by applicant): My research background includes: 1) investigating the transcriptional control of the gene 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. In applying for the NIH 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. This CDA will enable me to obtain additional training in several key areas that will facilitate my transition to an independent translational researcher. The research environment will include UCSF and three affiliated institutions: San Francisco General Hospital, the San Francisco VA, 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 (ART) by investigating whether there is ongoing replication in the gut, whether the gut is a major reservoir of latently-infected CD4+ T cells, and whether new antiretrovirals or anti- latency therapies can impact ongoing replication and/or latent infection in the gut. The specific aims are: 1) to compare HIV levels and immune parameters in different regions and cell types within the gut; 2) to assess for ongoing replication in the gut by measuring the effect of therapy intensification on levels of HIV, immune markers, and markers of abnormal gut barrier function; and 3) to assess the size of the gut latent reservoir, investigate mechanisms of latency in the gut, and assess the response of gut cells and tissues to anti-latency therapies. Study part 1 is a prospective, longitudinal pilot study in which participants will have baseline upper and lower endoscopy with biopsies from 4 regions followed by 12 weeks of intensification (with raltegravir +/- a second agent) and repeat endoscopies. Part 2 of the study will involve endoscopy with more intensive sampling from one gut site (part 2A) as well as unused tissue from HIV+ patients who are undergoing abdominal surgery for other reasons (2B). These biopsies and tissues will be used to identify the cell populations with the most HIV DNA and RNA, to establish culture systems 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. Though antiviral medicines have revolutionized the lives of those with HIV, these therapies are expensive, require lifelong administration and monitoring, have risk of side effects and toxicities, do not eradicate HIV, do not completely reverse the immune deficits of HIV, 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 or improved tolerance of lifelong HIV infection. This study seeks to improve understanding of how and why HIV persists on therapy by investigating whether there is persistent viral replication in the gut, whether the gut is a major reservoir of latently-infected cells, and whether new therapies can impact persistent replication and/or latent infection in the gut.