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: 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
• 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' 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

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.

项目总结 克罗恩病(CD)的特征是进行性的、跨壁的炎症，可影响到 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。 在他们的病程上。肛周瘘是一种发病率较高的CD并发症，治疗效果不佳。 非洲血统(AA)患者的肛周疾病，包括肛周瘘管，比欧洲人更高 先祖(EA)CD患者。全基因组关联时代的许多遗传进步都来自于研究 其中再生障碍性贫血患者的代表性不足。PTGER4是再生障碍性贫血中唯一具有强关联信号的风险基因座 人口。有必要将再生障碍性贫血患者纳入CD的遗传和基因组研究，特别是在 肛周瘘的背景，以更好地了解疾病的驱动因素和改善临床结果。 目的1旨在构建有肛周瘘管史的AA和EA患者的单细胞基因组学队列 CD.迄今为止，还没有针对直肠或瘘管的CD样本的单细胞测序数据集，也没有 CD中比较这些祖先群体之间单细胞基因表达的数据集。我们将执行CITE- 重度CD患者直肠和结肠切除术切除组织的序列分析 肛周瘘管。我们还将从活动性炎症区域和非受累直肠收集直肠活检。 或冒号用于单细胞RNA测序。从这些数据中，我们将识别差异基因表达模块 这将通过对来自更大CD患者队列的可用的批量RNA测序数据进行投影来验证。 单细胞捕获RNA和蛋白质表达将提供前所未有的细胞粒度 在瘘管进展中很重要的人群，并阐明患者人群之间的任何差异。 在目标2中，我们将进一步探讨瘘管发病的驱动因素。我们认为IL-17和PGE2信号是 炎症和上皮间质背景下影响上皮完整性的候选途径 过渡(EMT)，促进瘘管形成。我们将长期培养患者来源的上皮单层 用来模拟上皮分化、屏障功能和EMT的有机化合物。我们将使用这个系统来评估 前列腺素E_2和IL-17受体激动剂和抑制剂对上皮细胞的作用及其对创伤的影响 治愈和调解瘘管形成。通过这种方式，我们将能够询问特定的信号通路和 他们对上皮健康的贡献使用功能读数。 西奈山的学术环境是进行这项工作的无与伦比的环境。西奈山是拓荒者的故乡 在IBD临床护理和研究方面，他们以有意义的方式进行合作，同时与最多样化的 世界上最多的病人。这一奖学金的培训计划充分利用了这一优势。总之，这些研究 本文的描述将为肛周瘘的发生以及哪些基因模块可能导致 易受感染的病人。该方案将直接体外单细胞基因组学与体外功能研究相结合。 对影响不同患者群体的高发病率并发症进行强有力的调查。