Profiling intraepithelial lymphocyte populations in health and CrohnâÂÂs disease

分析健康和克罗恩病中的上皮内淋巴细胞群

• 批准号: 10017208
• 负责人: Karen Leigh Edelblum
• 金额: $ 19.88万
• 依托单位: RBHS-NEW JERSEY MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-12 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10017208
• 关键词: Acute; Affect; Area; Atlases; Biological Process; Biopsy; CD8B1 gene; Cells; Characteristics; Chronic; Collaborations; Crohn' s disease; Development; Disease; Disease remission; Distal part of ileum; Effector Cell; Endoscopic Biopsy; Enterocytes; Epithelial; Epithelium; Equilibrium; Evaluation; Excision; Exhibits; Fluorescent in Situ Hybridization; Frequencies; Future; Genetic; Goals; Health; Homeostasis; Human; Image; Immune; Immune response; Immunologic Surveillance; Immunologics; Individual; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Response; Intestinal Mucosa; Intestines; Knowledge; Lamina Propria; Lead; Licensing; Longitudinal Studies; Lymphocyte; Lymphocyte Function; Lymphocyte Subset; Lymphocytosis; Lymphoid Cell; Maintenance; Mission; Molecular Profiling; Mucosal Immunity; Mucous Membrane; National Institute of Diabetes and Digestive and Kidney Diseases; Operative Surgical Procedures; Pathogenesis; Patients; Phenotype; Population; Postoperative Period; Prognostic Marker; Property; Public Health; Publishing; RNA; Recurrence; Regulation; Reporting; Research; Sampling; Small Intestines; Surface; T-Lymphocyte; Technology; Testing; Therapeutic Intervention; Tissues; United States National Institutes of Health; Villus; Work; base; chronic inflammatory disease; cohort; cytotoxic; enteric pathogen; experimental study; functional plasticity; human disease; improved; insight; intraepithelial; microorganism; novel; novel diagnostics; novel marker; single molecule; single-cell RNA sequencing; targeted treatment; therapeutic development; transcriptome; transcriptomics

PROJECT SUMMARY. Immunological surveillance at barrier surfaces is essential to provide defense against enteric pathogens and maintain mucosal homeostasis. Disruption of the balance between pro-inflammatory and regulatory immune response can lead to a loss of mucosal tolerance and development of chronic inflammatory disease, such as Crohn's disease (CD). The majority of inflammatory responses in CD have been attributed to both innate and adaptive immune cells located in the lamina propria; however, relatively little is known regarding the contribution of the intraepithelial lymphocyte (IEL) compartment to CD pathogenesis. IELs, which include intraepithelial T cells (IET) and intraepithelial ILCs (IE-ILC), contribute to the maintenance of epithelial barrier integrity and function as first line of defense against luminal microorganisms. Due to the difficulty in obtaining a sufficient number of IELs from patient biopsies for in-depth functional analyses, we propose to leverage advances in single-cell RNA sequencing technology and RNA imaging to profile the phenotypic and functional characteristics of ileal IEL subpopulations in health and acute CD. Using transcriptomics as a guide to identify molecular signatures for individual IEL subsets, we will develop single-molecule fluorescence in situ hybridization (sm-FISH) probe multiplexes to visualize these cell populations in patient biopsies. In collaboration with the IBD Genetics Consortium, we will obtain tissue sections from longitudinal endoscopic biopsies of a CD patient cohort that has undergone ileal resection and determine the extent to which IEL phenotype, localization and effector function correlate to disease pathogenesis. These experiments will not only provide the first cell atlas of the human ileal IEL compartment, but may also lead to the development of novel prognostic biomarkers or therapeutic interventions for the treatment of CD.

项目摘要。 屏障表面的免疫监视对于防御肠道病毒至关重要 病原体并维持粘膜稳态。破坏促炎症之间的平衡 调节性免疫反应可能导致粘膜耐受性丧失和慢性病的发展 炎症性疾病，例如克罗恩病 (CD)。 CD 中的大部分炎症反应 归因于位于固有层的先天性和适应性免疫细胞；然而， 关于上皮内淋巴细胞 (IEL) 区室的贡献知之甚少 CD发病机制。 IEL，包括上皮内 T 细胞 (IET) 和上皮内 ILC (IE-ILC)， 有助于维持上皮屏障的完整性和作为第一道防线的功能 管腔微生物。由于很难从患者那里获得足够数量的 IEL 活检进行深入的功能分析，我们建议利用单细胞 RNA 的进步 测序技术和RNA成像来分析表型和功能特征 健康和急性 CD 中的回肠 IEL 亚群。使用转录组学作为识别分子的指南 针对各个 IEL 子集的特征，我们将开发单分子荧光原位杂交 (sm-FISH) 探针多重检测以可视化患者活检中的这些细胞群。合作中 与 IBD 遗传学联盟合作，我们将从纵向内窥镜活检中获得组织切片 接受回肠切除术的 CD 患者队列并确定 IEL 的程度 表型、定位和效应器功能与疾病发病机制相关。这些实验将 不仅提供了人类回肠 IEL 室的第一个细胞图谱，而且还可能导致 开发新的预后生物标志物或治疗 CD 的治疗干预措施。