Immunometabolic regulation of CD8+ T cell mediated intestinal epithelial cell death in people with HIV (PWH)

HIV 感染者 (PWH) 中 CD8 T 细胞介导的肠上皮细胞死亡的免疫代谢调节

• 批准号: 10528704
• 负责人: Douglas Kwon
• 金额: $ 70.8万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10528704
• 关键词: Address; Apoptosis; Autologous; Bacteria; Biological Assay; Biological Markers; Biological Models; Biopsy; Blood; Blood Circulation; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cell Death; Cell model; Cells; Cellular biology; Cessation of life; Chronic; Colon; Data; Defect; Development; Disease; Disease Progression; Endoscopy; Epithelial; Epithelial Cells; Functional disorder; Genetic Transcription; Gut associated lymphoid tissue; HIV; HIV Infections; Heart Diseases; Histology; Homeostasis; Human; Hyperglycemia; Immune; Immune System Diseases; Immunology; Impairment; In Situ; Individual; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Institutes; Intestinal Mucosa; Intestinal permeability; Intestines; Knowledge; Lipids; Massachusetts; Measures; Mediating; Membrane Lipids; Metabolic; Modeling; Molecular; Morbidity - disease rate; Mucous Membrane; Mus; Nutritional; Organoids; Pathogenesis; Pathologist; Pathology; Pathway interactions; Peripheral; Permeability; Persons; Phenotype; Population; Positioning Attribute; Process; Regimen; Regulation; Role; Sample Size; Sampling; Stroke; T-Lymphocyte; Technology; Testing; Tissue Sample; Tissues; Virus Diseases; Virus Replication; antiretroviral therapy; base; cohort; exhaustion; fatty acid metabolism; gastrointestinal; gastrointestinal epithelium; gut bacteria; gut inflammation; immune activation; improved; improved outcome; in vivo; in vivo Model; intestinal barrier; intestinal epithelium; microbial; mortality; mouse model; novel; self-renewal; single cell sequencing; systemic inflammatory response; transcriptome sequencing

A hallmark of human immunodeficiency virus (HIV) infection is intestinal inflammation and impairment of gut epithelial barrier function. These defects are thought to drive HIV disease progression by allowing translocation of luminal microbial products into the circulation, which triggers chronic systemic immune activation and disease progression. Although antiretroviral therapy (ART) effectively suppresses viral replication in the blood, it does not restore homeostasis in the intestine, even after years of treatment. This contributes to increased morbidity and mortality due to inflammatory non-communicable diseases (NCDs) such as heart disease and stroke in people with HIV (PWH). Despite the central role of intestinal barrier function in HIV disease pathogenesis, little is known about the specific mechanisms by which HIV results in epithelial damage. Studies of intestinal epithelial cell (IEC) dysfunction in HIV have relied primarily on correlative observations based on histology or peripheral biomarkers. Our proposal addresses critical gaps in knowledge by utilizing intestinal tissue samples from well-characterize cohorts of PWH and HIV-uninfected individuals, along with novel in vivo mouse and ex vivo human cell models and state-of-the-art technologies to explore specific mechanisms by which HIV contributes to IEC death in PWH. We hypothesize that HIV- associated defects in CD8+ T cell fatty acid (FA) metabolism cause them to scavenge lipids from nearby IEC, which leads to IEC death. This IEC death then results in intestinal barrier disruption. To address these hypotheses, we are proposing two complementary aims. In Aim 1 we will characterize the in vivo and ex vivo impact of HIV infection on IEC death in PWH using colon tissue samples obtained by endoscopy to characterize IEC death and CD8+ T cell phenotypes. We will also utilize novel mini-gut organoid models that incorporate autologous tissue resident immune cells and in-depth single cell sequencing analysis leveraging a platform optimized for small sample sizes developed by collaborators at MIT. In Aim 2, we will determine the mechanisms by which HIV-associated dysregulation of FA metabolism in intestinal CD8+ T cells disrupts the colonic epithelial barrier. In this aim we will utilize ex vivo human organoid and in vivo murine models to test the mechanistic role of impaired FA metabolism in CD8+ T cell mediated IEC death and intestinal barrier dysfunction. This approach will identify specific cellular and molecular mechanisms underlying the impact of HIV infection on intestinal barrier function and systemic immune activation. To carry out these Aims we have assembled a team with combined expertise in HIV disease, mucosal immunology, and GI pathology, who are well positioned to uncover specific mechanisms that underlie intestinal epithelial dysfunction in HIV infection. This proposal will address important unknown mechanisms of IEC biology that may help in the development of new strategies to reverse gut barrier defects in HIV infection.

人类免疫缺陷病毒（HIV）感染的标志是肠炎和障碍 肠上皮屏障功能。这些缺陷被认为可以通过允许 腔体微生物产物转运到循环中，这会触发慢性全身免疫 激活和疾病进展。尽管抗逆转录病毒疗法有效抑制病毒 在血液中的复制，即使经过多年的治疗，它也不会恢复肠内稳态。这 导致由于炎症性非传染性疾病（NCD）而引起的发病率和死亡率的增加 例如艾滋病毒（PWH）患者的心脏病和中风。尽管肠壁的核心作用 在HIV疾病发病机理中的功能，对HIV导致的特定机制知之甚少 上皮损害。 HIV中肠上皮细胞（IEC）功能障碍的研究主要依赖于 基于组织学或外围生物标志物的相关观察结果。我们的建议解决了关键的差距 通过利用PWH和HIV未直发的肠道组织样品利用肠道组织样品 个人，新型体内小鼠和体内人体细胞模型以及最先进的技术 探索艾滋病毒在PWH中为IEC死亡做出贡献的特定机制。我们假设HIV- CD8+ T细胞脂肪酸（FA）代谢的相关缺陷导致它们从附近清除脂质 IEC，导致IEC死亡。然后，IEC死亡会导致肠道障碍。解决这些 假设，我们提出了两个补充目标。在AIM 1中，我们将表征体内和Ex Vivo HIV感染对通过内窥镜获得的结肠组织样本对PWH的IEC死亡的影响 表征IEC死亡和CD8+ T细胞表型。我们还将利用新型迷你器官模型 结合自体组织驻留免疫细胞和深入的单细胞测序分析 利用MIT合作者开发的小样本量优化的平台。在AIM 2中，我们将 确定肠道CD8+ T中FA代谢失调的机制 细胞破坏结肠上皮屏障。在此目标中，我们将利用体内人体器官和体内鼠 测试FA代谢受损的机械作用在CD8+ T细胞中介导的IEC死亡和 肠屏障功能障碍。这种方法将识别特定的细胞和分子机制 HIV感染对肠道屏障功能和全身免疫激活的影响。执行这些 目的我们已经组建了一个在艾滋病毒疾病，粘膜免疫学和GI方面具有联合专业知识的团队 病理学，他们良好地揭示了肠上皮功能障碍的特定机制 在HIV感染中。该建议将解决IEC生物学重要的未知机制，这可能有助于 制定逆转肠道屏障缺陷的新策略。