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.

项目概要 克罗恩病 (CD) 的特点是进行性、透壁性炎症，可影响身体的任何区域。 胃肠道。高达 50% 的 CD 患者会出现穿透性并发症，最常见的是肛周瘘 他们的病程。肛周瘘管是一种高发病率的 CD 并发症，且治疗效果不佳。 非洲血统 (AA) 患者的肛周疾病（包括肛周瘘管）发病率高于欧洲血统患者 祖先 (EA) CD 患者。全基因组关联时代的许多遗传进展都来自研究 其中 AA 患者代表性不足。 PTGER4 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中，特别是在 肛周瘘的背景，以更好地了解疾病的驱动因素并改善临床结果。 目标 1 旨在构建一个由有肛周瘘病史的 AA 和 EA 患者组成的单细胞基因组队列 光盘。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 比较 CD 中这些祖先群体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 且有病史的患者进行直肠切除术和结肠切除术的切除组织进行测序 肛周瘘。我们还将从活动性炎症区域和未受累的直肠收集直肠活检组织 或结肠用于单细胞 RNA 测序。从这些数据中，我们将识别差异基因表达模块 这将通过投影到来自更大 CD 患者队列的可用批量 RNA 测序数据来验证。 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 人群对瘘管病进展很重要，并阐明了患者人群之间的差异。 在目标 2 中，我们将进一步探讨瘘管发病机制的驱动因素。我们将 IL-17 和 PGE2 信号传导确定为 在炎症和上皮间充质背景下影响上皮完整性的候选途径 过渡（EMT），促进瘘管发生。我们将从患者来源长期培养上皮单层 类器官来模拟上皮分化、屏障功能和 EMT。我们将使用该系统来评估 PGE2 和 IL-17 受体激动和抑制对上皮细胞的影响，以及这如何影响伤口 愈合并介导瘘管的发生。通过这种方式，我们将能够询问特定的信号传导途径并 他们使用功能读数对上皮健康的贡献。 西奈山的学术环境对于开展这项工作来说是无与伦比的。西奈山是先驱者的故乡 在 IBD 临床护理和研究领域，他们以有意义的方式合作，同时与最多元化的人之一合作 世界上的患者人数。该奖学金的培训计划充分利用了这一点。一起，研究 本文描述的内容将提供对肛周瘘起始以及哪些基因模块可能产生的重要见解。 易感患者。该提案将直接离体单细胞基因组学与体外功能研究相结合 对影响不同患者群体的高发病率并发症进行深入调查。