Quantification of HIV-1 Reservoirs in the Gut

简介(申请人提供)：我的研究背景包括：1)研究c-kit基因的转录调控；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。通过申请美国国立卫生研究院职业发展奖(CDA)，我希望获得必要的经验、技能和知识，成为一名独立的教职员工调查员，利用患者驱动的研究更好地了解根除艾滋病毒的障碍。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。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、旧金山退伍军人管理局和格莱斯顿研究所。这些机构以艾滋病毒的临床护理、培训和研究而闻名全国。这项研究旨在通过调查肠道中是否有正在进行的复制，肠道是否是潜伏感染的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疗法是否会影响肠道中正在进行的复制和/或潜伏感染，来提高对HIV持续接受抗逆转录病毒治疗(ART)的方式和原因的理解。其具体目标是：1)比较肠道内不同区域和细胞类型的艾滋病毒水平和免疫参数；2)通过测量强化治疗对艾滋病毒、免疫标志物和肠道屏障功能异常标志物水平的影响，评估艾滋病毒在肠道内的持续复制；以及3)评估肠道潜伏库的大小，研究肠道潜伏期的机制，以及评估肠道细胞和组织对抗潜伏期治疗的反应。研究第一部分是一项前瞻性的纵向先导性研究，参与者将接受基线的上下内窥镜检查，从4个区域进行活组织检查，然后进行12周的强化(使用雷替格雷+/-第二种药物)和重复的内窥镜检查。研究的第二部分将涉及内窥镜检查，从一个肠道部位(第2A部分)进行更密集的采样，以及从因其他原因接受腹部手术的HIV+患者的未使用组织(2B)。这些活检和组织将被用来识别拥有最多艾滋病毒DNA和RNA的细胞群体，建立培养系统来测量肠道中可诱导和/或复制能力的艾滋病毒，研究肠道潜伏的机制，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。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，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，很难提供给每个艾滋病毒感染者。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或改善对终身艾滋病毒感染的耐受性的方法。这项研究旨在通过调查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，以及新的治疗方法是否可以影响肠道的持续复制和/或潜伏感染，来提高对艾滋病毒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.