Defining mechanisms of perianal fistula inception in African and European ancestry Crohn's disease patients

非洲和欧洲克罗恩病患者肛周瘘起始的定义机制

基本信息

批准号：
10314518
负责人：
Rachel Levantovsky
金额：
$ 4.59万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10314518
关键词：
Adopted Affect African Agonist Air Area Biological Assay Biopsy Cell physiology Cells Cellular Indexing of Transcriptomes and Epitopes by Sequencing Clinical Clinical Research Colectomy Colon Complement Complication Crohn&apos s disease Data Data Set Development Dinoprostone Disease Electrical Resistance Environment Epithelial Epithelial Cells Epitopes Etiology European Event Excision Fellowship Fibrosis Fistula Fostering Functional disorder Gastrointestinal tract structure Gene Expression Gene Expression Profile Gene Expression Profiling Genes Genetic Genetic Risk Genetic Transcription Genomics Genotype Goals Health Home Immune Immunofluorescence Immunologic In Vitro Inflammation Inflammatory Interleukin-17 Intervention Intestines Lipids Liquid substance Maintenance Measures Mediating Mediator of activation protein Mesenchymal Modeling Morbidity - disease rate Mucous Membrane Organoids Outcome Pathogenesis Pathologic Pathology Pathway interactions Patient-Focused Outcomes Patients Pattern Phenotype Physiological Play Polymerase Chain Reaction Population Proteins RNA Recording of previous events Rectum Risk Role Sampling Signal Pathway Signal Transduction Specimen Stains Stimulus Stromal Cells System TNF gene Tight Junctions Time Tissue-Specific Gene Expression Tissues Training Translations Variant Visualization WFDC2 gene Work ancestry analysis base behavior influence chronic inflammatory disease clinical care cohort cytokine data acquisition epithelial injury genetic variant genome wide association study genome-wide healing improved in vivo indexing insight interleukin-17C intestinal epithelium loss of function monolayer novel patient population power analysis protein expression receptor recruit rectal risk variant single cell sequencing single-cell RNA sequencing transcriptome transcriptome sequencing treatment response validation studies wound wound healing

项目摘要

PROJECT SUMMARY Crohn’s disease (CD) is characterized by progressive, transmural inflammation that can affect any region of the gastrointestinal tract. Penetrating complications, most often perianal fistulae, will affect up to 50% of CD patients over their disease course. Perianal fistulae are a high morbidity CD complication with poor treatment response. African ancestry (AA) patients have higher rates of perianal disease, including perianal fistula, than European ancestry (EA) CD patients. Many of the genetic advances of the genome wide association era are from studies wherein AA patients are underrepresented. PTGER4 is the only risk locus with strong association signals in AA populations. There is a need to include AA patients in genetic and genomic studies of CD, particularly in the context of perianal fistula, to better understand drivers of disease and improve clinical outcomes. Aim 1 seeks to construct a single cell genomics cohort of AA and EA patients with a history of perianal fistulizing CD. To date, there is no single cell sequencing dataset of CD samples with a focus on the rectum or fistula, nor a dataset comparing single cell gene expression between these ancestry groups in CD. We will perform CITE- sequencing on resection tissues from proctectomies and colectomies in patients with severe CD and history of perianal fistula. We will also collect rectal biopsies from areas of active inflammation and from un-involved rectum or colon for single cell RNA sequencing. From these data, we will identify differential gene expression modules that will be validated by projection onto available bulk RNA sequencing data from larger CD patient cohorts. Single cell capture of both RNA and protein expression will provide unprecedented granularity into cell populations important in fistula progression and illuminate any differences between patient populations. In Aim 2, we will further explore drivers of fistula pathogenesis. We identify IL-17 and PGE2 signaling as candidate pathways that influence epithelial integrity in the context of inflammation and epithelial-mesenchymal transition (EMT), promoting fistula inception. We will culture epithelial monolayers long-term from patient-derived organoids to model epithelial differentiation, barrier function, and EMT. We will use this system to assess the effects of PGE2 and IL-17 receptor agonism and inhibition on epithelial cells, and how this may influence wound healing and mediate fistula inception. In this way, we will be able to interrogate specific signaling pathways and their contributions to epithelial health using functional readouts. The academic environment at Mount Sinai is unparalleled to perform this work. Mount Sinai is home to pioneers in IBD clinical care and research, who collaborate in meaningful ways while working with one of the most diverse patient populations in the world. The training plan for this fellowship takes full advantage. Together, the studies described herein will provide important insight into perianal fistula inception and which gene modules might make a patient susceptible. This proposal combines direct ex vivo single cell genomics with in vitro functional studies for a robust inquiry into a high morbidity complication that affects diverse patient populations.

项目摘要克罗恩病（CD）的特征是进行性透壁炎症，可能影响胃肠道。穿透并发症，最常见的并发症，最多会影响50％的CD患者在他们的疾病过程中。骨瘘是高发病率CD并发症，治疗反应不佳。非洲血统（AA）患者的验收率较高，包括perian瘘，而不是欧洲祖先（EA）CD患者。基因组广泛关联时代的许多遗传进步来自研究其中AA患者的代表性不足。 PTGER4是AA中唯一具有较强关联信号的风险基因座人群。需要将AA患者纳入CD的遗传和基因组研究，特别是在 perian瘘的背景，以更好地了解疾病的驱动因素并改善临床结果。 AIM 1旨在构建具有Perianal Fistulsing病史的AA和EA患者的单个细胞基因组学队列光盘。迄今一个数据集比较CD中这些祖先组之间的单细胞基因表达。我们将执行引用 - 严重CD患者的凝乳切除术和结肠切除术的切除组织的测序以及病史 Perian瘘。我们还将从主动感染和未涉及直肠的区域收集直肠活检或用于单细胞RNA测序的结肠。从这些数据中，我们将确定差异基因表达模块这将通过投影对来自较大CD患者队列的可用大量RNA测序数据进行验证。 RNA和蛋白质表达的单细胞捕获将为细胞提供前所未有的粒度人群在瘘管进展中很重要，并照亮了患者人群之间的任何差异。在AIM 2中，我们将进一步探索瘘管发病机理的驱动因素。我们将IL-17和PGE2信号识别为在感染和上皮间质的背景下影响上皮完整性的候选途径过渡（EMT），促进瘘管的启动。我们将长期从患者衍生的长期培养上皮单层器官以建模上皮分化，屏障功能和EMT。我们将使用该系统评估 PGE2和IL-17受体激动剂和抑制对上皮细胞的影响，以及这可能如何影响伤口治愈和介导瘘管的启动。这样，我们将能够询问特定的信号通路和他们使用功能读数对上皮健康的贡献。西奈山的学术环境无法执行这项工作。西奈山是开拓者的家在IBD临床护理和研究中，他们在与最潜水员之一合作的同时以有意义的方式合作世界上的患者人口。该奖学金的培训计划充分利用了。一起研究本文描述的将提供有关perianal瘘管的重要见解，以及哪些基因模块可能会产生患者易感性。该建议将直接的离体单细胞基因组与体外功能研究结合在一起为了对高发病并发症进行强有力的调查，影响潜水者的患者人群。