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

• 批准号: 10649458
• 负责人: Rachel Levantovsky
• 金额: $ 5.27万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 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/10649458
• 关键词: Adopted; Affect; African ancestry; Agonist; Air; Anal Fistula; Anus; Area; Biological Assay; Biopsy; Cell physiology; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Clinical; Clinical Research; Cohort Studies; Colectomy; Collaborations; Colorectal; Complement; Complication; Crohn' s disease; Data; Data Set; Development; Dinoprostone; Disease; Disparate; Electrical Resistance; Environment; Epithelial Cells; Epithelium; Epitopes; Etiology; European ancestry; 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; IL17 gene; Immune; Immunofluorescence Immunologic; In Vitro; Inflammation; Inflammatory; Intervention; Intestines; Lipids; Liquid substance; Maintenance; Measures; Mediating; Mediator; Mesenchymal; Modeling; Morbidity - disease rate; Mucous Membrane; Organoids; Outcome; Pathogenesis; Pathologic; Pathology; Pathway interactions; Patient-Focused Outcomes; Patients; Pattern; Penetration; Phenotype; Physiological; Play; Polymerase Chain Reaction; Population; Predisposition; Proteins; RNA; Receptor Inhibition; 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; behavior influence; chronic inflammatory disease; clinical care; cohort; cytokine; data acquisition; epithelial injury; genetic variant; genome wide association study; genome-wide; genome-wide analysis; 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

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、透壁性炎症，可影响全身任何部位。 胃肠道穿透性并发症，最常见的是肛周瘘，将影响高达50%的CD患者 在他们的疾病过程中。肛周瘘是一种高发病率的CD并发症，治疗效果差。 非洲血统（AA）患者肛周疾病（包括肛周瘘）的发生率高于欧洲血统（AA）患者。 祖先（EA）CD患者。全基因组关联时代的许多遗传学进展都来自研究 其中AA患者代表性不足。PTGER 4是AA中唯一具有强关联信号的风险位点 人口。有必要将AA患者纳入CD的遗传和基因组研究，特别是在 在肛周瘘的背景下，更好地了解疾病的驱动因素并改善临床结果。 目的1：建立一个有肛周瘘管病史的AA和EA患者的单细胞基因组学队列 CD.到目前为止，还没有CD样本的单细胞测序数据集，重点是直肠或瘘管， 比较CD中这些祖先组之间的单细胞基因表达的数据集。我们将执行CITE- 对患有严重CD且有以下病史的患者的直肠切除术和结肠切除术的切除组织进行测序 肛周瘘管我们还将收集活动性炎症区域和未受累直肠的直肠活检标本 或结肠进行单细胞RNA测序。从这些数据中，我们将确定差异基因表达模块 这将通过投影到来自较大CD患者队列的可用批量RNA测序数据上来验证。 RNA和蛋白质表达的单细胞捕获将为细胞内提供前所未有的粒度。 患者人群在瘘管进展中的重要性，并阐明患者人群之间的任何差异。 在目标2中，我们将进一步探索瘘管发病机制的驱动因素。我们确定IL-17和PGE 2信号传导为 在炎症和上皮-间质细胞间质分化背景下影响上皮完整性的候选途径 过渡（EMT），促进瘘管病的发生。我们将从患者来源的细胞中长期培养上皮单层， 类器官用于模拟上皮分化、屏障功能和EMT。我们将使用这个系统来评估 PGE 2和IL-17受体激动和抑制对上皮细胞的影响，以及这可能如何影响伤口 愈合和介导瘘管形成。通过这种方式，我们将能够询问特定的信号通路， 它们对上皮健康的贡献。 在西奈山的学术环境是无与伦比的执行这项工作。西奈山是拓荒者的家园 在IBD临床护理和研究中，他们以有意义的方式合作，同时与世界上最多样化的 世界上的病人。该研究金的培训计划得到了充分利用。总之，这些研究 本文所述的将提供重要的洞察肛周瘘管发病和哪些基因模块可能使 易受感染的病人该建议将直接的离体单细胞基因组学与体外功能研究相结合 对影响不同患者人群的高发病率并发症进行有力调查。