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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要全世界 90% 以上的 HIV 感染都是通过粘膜途径发生的。大量 CD4 T 细胞耗竭、病毒感染、肠上皮细胞凋亡、上皮紧密连接破坏导致微生物易位和淋巴组织纤维化的 SIV 感染恒河猴 (RM) 模型。被认为是研究 HIV 相关肠病的公认模型，我们最近发现 SIV 感染后不久就会发生肠上皮细胞 (iEC) 凋亡和 iEC 再生减少。 IL-10R 的内化对 IL-10 信号传导和 IL-10 介导的失调产生影响抗炎反应在 iEC 损伤和随后的 SIV 发病机制中也发挥着至关重要的作用。了解 HIV/SIV 中异常的 iEC 再生可能会带来一种替代方法基于增强小肠 iEC 修复机制治疗 HIV 介导的肠病，每个肠绒毛都被至少六个 Lieberkühn 隐窝包围，其中容纳了专门的自我更新的干细胞和祖细胞，以及产生各种分化的 LGR5+（Leu-）。含有G蛋白偶联受体5)的丰富重复序列被鉴定为隐窝基底柱状的标记 (CBC) 在成体肠隐窝基部选择性表达的干细胞的遗传分析。 LGR5+肠干细胞（ISC）揭示单个干细胞可导致成人隐窝稳态干细胞通过克隆扩增产生极化肠上皮细胞、杯状细胞、肠内分泌细胞。细胞和簇细胞在经过几个瞬时扩增 (TA) 阶段后迁移到绒毛。 TA 细胞最终还分化为潘氏细胞 (PC)，这是一种高度特化的分泌细胞类型，最近的细胞消融和共培养散布在小肠隐窝底部的干细胞。研究表明 PC 提供必要的生态位信号来支持 LGR5+ 干细胞更新和存活。因此，我们目睹了 HIV/SIV 感染破坏了 LGR5+ ISC 和 LGR5+ ISC 之间的通讯。 PC 导致 iEC 再生减少在目标 1 中，我们将定义 LGR5+ 干细胞的相关性。在目标 2 中，我们将发现 ISC 和 PC 的变化。在两个子目标 2a 中，我们将确定 SIV 感染中的转录。在正常和 SIV 感染的 RM 中对 ISC 和 PC 进行编程在子目标 2b 中，我们将比较菌落。 SIV感染和正常RM之间恒河猴肠道LGR5+干细胞的形成效率。