Impact of HIV/SIV Infection on Paneth and Intestinal Stem Cell Interaction

HIV/SIV 感染对潘氏和肠道干细胞相互作用的影响

• 批准号: 9349058
• 负责人: Bapi Pahar
• 金额: $ 63.55万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-05 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9349058
• 关键词: ALCAM gene; Abbreviations; Ablation; Acquired Immunodeficiency Syndrome; Acute; Adult; Animal Model; Anti-inflammatory; Apoptosis; Bacteria; Biopsy; CD4 Positive T Lymphocytes; CD44 gene; Cell Communication; Cell Compartmentation; Cell physiology; Cells; Chronic; Clonal Expansion; Coculture Techniques; Communication; Data; Development; EGF gene; Enterocytes; Enteroendocrine Cell; Epidermal Growth Factor; Epithelial; Epithelial Cells; Family; Fibrosis; G-Protein-Coupled Receptors; Gastrointestinal Diseases; Gastrointestinal tract structure; Generations; Genes; Genetic Transcription; Goblet Cells; HIV; HIV Enteropathy; HIV Infections; Homeostasis; Impairment; In Vitro; Individual; Infection; Inflammatory Response; Injury; Interferons; Interleukin-10; Intestines; LGR5 gene; Lead; Leucine-Rich Repeat; Ligands; Link; Lymphoid Tissue; Macaca; Macaca mulatta; Mediating; Modeling; Molecular; Mouse Mammary Tumor Virus; Natural regeneration; Paneth Cells; Pathogenesis; Pathology; Patients; Play; Positioning Attribute; Quantitative Reverse Transcriptase PCR; Recovery; Regulator Genes; Reserve Stem Cell; Role; Route; SIV; Secretory Cell; Signal Transduction; Site; Small Intestines; Stem cells; Structure of intestinal gland; Study models; Systems Biology; Testing; Tight Junctions; Tissues; Transforming Growth Factors; Villus; Virus Diseases; base; cell injury; cell type; differential expression; genetic analysis; human tissue; immune activation; integration site; interleukin-10 receptor; intestinal crypt; jejunum; microbial; nonhuman primate; notch protein; protein expression; repaired; selective expression; self renewing cell; stem cell division; stem cell population; transcriptome; transcriptomics

PROJECT SUMMARY Worldwide more than 90% of all HIV infections occur via mucosal route. The gastrointestinal tract is a site of massive CD4 T-cell depletion, viral infection, enterocyte apoptosis, disruption of epithelial tight junctions leading to microbial translocation, and lymphoid tissue fibrosis. The SIV infected rhesus macaque (RM) model is considered a well-accepted model to study of HIV-associated enteropathy and we have recently discovered that intestinal epithelial cell (iEC) apoptosis and diminished iEC regeneration occur soon after SIV-infection. Internalization of IL-10R with the resultant impact on IL-10 signaling and dysregulation of the IL-10-mediated anti-inflammatory responses also play a crucial role in iEC damage and subsequent SIV pathogenesis. Understanding the aberrant iEC regeneration in HIV/SIV might lead to an alternative approach for the treatment of HIV-mediated enteropathy based on enhancing iEC repair mechanisms. In the small intestine, each intestinal villus is encircled by at least six crypts of Lieberkühn, which house dedicated populations of stem and progenitor cells that self-renew, as well as give rise to the various differentiated iECs. LGR5+ (Leu- rich repeat containing G protein-coupled receptor 5) was identified as the marker for crypt base columnar (CBC) stem cells that was selectively expressed at the base of adult intestinal crypts. Genetic analysis of LGR5+ intestinal stem cells (ISCs) revealed that a single stem cell leads to crypt homeostasis in adult intestines by clonal expansion. Stem cells give rise to the polarized enterocytes, goblet cells, enteroendocrine cells, and tuft cells that migrate up to the villi after passing through several transiently-amplifying (TA) stages. TA cells also terminally differentiate into Paneth cells (PCs), a highly specialized secretory cell type which intersperse with stem cells at the base of the crypts in the small intestine. Recent cell ablation and co-culture studies showed that PCs provide essential niche signals to support LGR5+ stem cell renewal and survival. Therefore, we hypothesize that HIV/SIV infection disrupts the communication between LGR5+ ISC and PCs leading to diminished iECs regeneration. In Aim 1, we will define the correlates of LGR5+ stem cells and PCs with SIV/HIV infection status. In Aim 2, we will uncover the alterations of ISCs and PCs communication in SIV infection in two sub aims. In Sub-Aim 2a, we will determine the transcriptional programming of ISCs and PCs in normal and SIV infected RMs. In Sub-Aim 2b, we will compare the colony forming efficiency of rhesus intestinal LGR5+ stem cells between SIV infected and normal RMs.

项目总结 在世界范围内，90%以上的艾滋病毒感染是通过粘膜途径发生的。胃肠道是一种 大量的CD4T细胞耗尽，病毒感染，肠细胞凋亡，上皮紧密连接中断 导致微生物移位和淋巴组织纤维化。SIV感染恒河猴(RM)模型 被认为是研究HIV相关肠病的一个被广泛接受的模型，我们最近发现 SIV感染后不久，肠上皮细胞(IEC)发生凋亡，IEC再生减少。 IL-10R内化对IL-10信号转导的影响及IL-10介导的调节失调 抗炎反应也在IEC损伤和随后的SIV发病机制中发挥关键作用。 了解HIV/SIV中IEC的异常再生可能会导致另一种治疗方法 基于增强IEC修复机制的HIV介导肠病的治疗。在小肠里， 每个肠道绒毛被至少六个利伯库恩地穴环绕，这些地穴容纳着专门的种群 干细胞和祖细胞自我更新，以及产生各种分化的IECs。Lgr5+(亮氨酸- 富含G蛋白偶联受体5的重复序列)可作为隐窝基柱的标志 (CBC)在成人肠隐窝底部选择性表达的干细胞。遗传病的分析 Lgr5+肠道干细胞(ISCs)研究表明，单个干细胞可导致成人隐窝的动态平衡 通过克隆扩张的肠道。干细胞产生极化的肠上皮细胞、杯状细胞、肠内分泌 细胞和丛生细胞，在经过几个瞬时放大(TA)阶段后迁移到绒毛。 TA细胞也终末分化为Paneth细胞(PC)，这是一种高度专业化的分泌细胞类型， 在小肠的隐窝底部点缀着干细胞。最近的细胞消融和共培养 研究表明，PC提供了支持LGR5+干细胞更新和存活所必需的利基信号。 因此，我们假设HIV/SIV感染扰乱了LGR5+ISC和LGR5+ISC之间的通信 导致IECS再生减弱的PC。在目标1中，我们将定义LGR5+干细胞的相关性 以及具有SIV/HIV感染状态的PC。在目标2中，我们将揭示ISC和PC的变化 传播在SIV感染中的两个分目标。在次级目标2a中，我们将确定转录 正常和SIV感染的RMS中的ISCs和PC的编程。在子目标2b中，我们将比较 SIV感染与正常RMS间恒河猴肠道LGR5+干细胞的形成效率