Regulatory role for an epithelial repair protein, LINGO2, in colitis

上皮修复蛋白 LINGO2 在结肠炎中的调节作用

基本信息

批准号：
10039666
负责人：
Nicole Maloney Belle
金额：
$ 16.84万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10039666
关键词：
3-Dimensional Abscess Affect Azoxymethane Biological Models Biology Bone Marrow CRISPR/Cas technology Carcinogens Carcinoma Cells Cellular biology Chimera organism Chronic Colitis Colitis associated colorectal cancer Colonic Neoplasms Data Defect Development Disease Disease model EGFR inhibition Environment Epidermal Growth Factor Receptor Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor Epithelial Epithelial Cells Epithelium Exposure to Faculty Fistula Foundations Functional disorder Funding Genes Genetic Genetic Polymorphism Goals Growth Hematopoietic Hemorrhagic colitis Homeostasis Human IL6 gene Immune Immunology In Vitro Inflammation Inflammatory Inflammatory Bowel Diseases Inflammatory Response Interleukin-6 Intestines Investigation Kinetics Knock-in Knock-out Knockout Mice Leucine-Rich Repeat Malignant - descriptor Malignant Neoplasms Modeling Molecular Mucous Membrane Mus Mutant Strains Mice Myelogenous Myeloid Cells Organoids Pathway interactions Patients Pennsylvania Phosphorylation Predictive Factor Predisposition Production Protein Family Proteins Publications Publishing RIPK1 gene Regulation Relapse Research Risk Role STAT3 gene Severities Signal Transduction Sodium Dextran Sulfate Susceptibility Gene Techniques Testing Training Transplantation United States National Institutes of Health Universities Up-Regulation adenoma career development colitis associated cancer cytokine dextran sulfate sodium induced colitis experimental study genetic manipulation genome wide association study inflammatory disease of the intestine innovation intestinal barrier intestinal epithelium knowledge base leucine-rich repeat protein member microbiota mouse model novel overexpression receptor repaired response skills small molecule tenure track tool trefoil factor tumor tumor microenvironment tumorigenic

项目摘要

PROJECT SUMMARY Inflammatory bowel disease (IBD) constitutes a spectrum of incurable, relapsing and remitting disorders driven by intestinal mucosal barrier function defects and aberrant pro-inflammatory responses directed against microbial flora. IBD patients are at increased risk of developing colitis-associated colorectal cancer (CAC) but the epithelial specific mechanisms that control it are incompletely understood. Also unclear are the genetic factors that predict which IBD patients are likely to progress to CAC. The research focus of this proposal is to uncover how dysregulation of a novel epithelial repair protein, Leucine rich repeat and Ig domain-containing, nogo receptor-interacting protein family 2 (LINGO2) contributes to malignant transformation. In preliminary data, we show that Lingo2 deficiency in epithelial cells results in up-regulation of EGFR phosphorylation, prolonged STAT3 activation in response to IL6 treatment and increased proliferation compared to wildtype cells. Furthermore, Lingo2 deficient mice have severe dextran sodium sulfate (DSS) induced colitis that is rescued by EGFR inhibition. We demonstrate that Lingo2 is expressed in immune cells and its loss in the hematopoietic compartment contributes to susceptibility to colitis. Consistent with the published observations that increased EGFR signaling is pro-tumorigenic, we show that when exposed to azoxymethane (AOM) and DSS, Lingo2 deficient mice are more susceptible to the development of CAC compared to control mice. I therefore hypothesize that Lingo2 protects against the development of CAC through regulation of EGFR and STAT3 activity. The objective is to first mechanistically dissect the cell autonomous role of Lingo2 in epithelial cells in protection from CAC and then to understand whether Lingo2 deficiency in the microenvironment is important for CAC. My hypothesis will be tested through two inter-related Specific Aims that will examine the interplay of LINGO2 and EGFR signaling in epithelial cells in CAC (Aim 1) and then evaluate how Lingo2 deficiency affects the production of STAT3 activating cytokines and activity of myeloid cells in CAC (Aim 2). The experiments proposed will use various innovative approaches including the use of unique knock-in murine models, 3D organoid culture model systems and organoid orthotopic transplantation. The proposed research is significant because it will provide new information about a novel tumor suppressive molecular pathway. University of Pennsylvania provides the perfect research environment to conduct this investigation given local expertise in the inter-related fields of mucosal biology, epithelial biology and immunology. The candidate will gain fundamental skills in murine and in vitro disease modeling and broaden her immunology and epithelial biology knowledge base. These skill sets will greatly enhance the candidate's career development into an independent NIH funded tenure-track faculty member with the long term goal of understanding the mechanisms that control mucosal repair and homeostasis.

项目概要炎症性肠病 (IBD) 是一系列无法治愈、复发和缓解的疾病肠粘膜屏障功能缺陷和异常的促炎反应直接针对微生物菌群。 IBD 患者患结肠炎相关结直肠癌 (CAC) 的风险增加，但控制它的上皮特异性机制尚不完全清楚。遗传也不清楚预测哪些 IBD 患者可能进展为 CAC 的因素。本提案的研究重点是揭示一种新型上皮修复蛋白（富含亮氨酸重复序列和含有 Ig 结构域）的失调， nogo 受体相互作用蛋白家族 2 (LINGO2) 有助于恶性转化。在初步数据显示，上皮细胞中的 Lingo2 缺陷导致 EGFR 磷酸化上调，与野生型相比，IL6 治疗响应的 STAT3 激活时间延长，增殖增加细胞。此外，Lingo2 缺陷小鼠患有严重的右旋糖酐硫酸钠 (DSS) 诱导的结肠炎，即 EGFR 抑制可挽救。我们证明 Lingo2 在免疫细胞中表达，并且在免疫细胞中表达缺失造血室有助于结肠炎的易感性。与已发表的观察结果一致增加的 EGFR 信号传导具有促肿瘤发生作用，我们表明，当暴露于氧化偶氮甲烷 (AOM) 和与对照小鼠相比，DSS、Lingo2 缺陷小鼠更容易发生 CAC。我因此假设 Lingo2 通过调节 EGFR 和 STAT3 活性。目的是首先从机制上剖析 Lingo2 在上皮细胞中的细胞自主作用细胞免受 CAC 侵害，然后了解微环境中的 Lingo2 缺陷是否与对于 CAC 来说很重要。我的假设将通过两个相互关联的具体目标进行检验，这两个目标将检验 LINGO2 和 EGFR 信号在 CAC 上皮细胞中的相互作用（目标 1），然后评估 Lingo2 如何发挥作用缺乏会影响 CAC 中 STAT3 激活细胞因子的产生和骨髓细胞的活性（目标 2）。拟议的实验将使用各种创新方法，包括使用独特的敲入小鼠模型、3D 类器官培养模型系统和类器官原位移植。拟议的研究是意义重大，因为它将提供有关新型肿瘤抑制分子途径的新信息。鉴于当地情况，宾夕法尼亚大学为开展这项调查提供了完美的研究环境粘膜生物学、上皮生物学和免疫学等相关领域的专业知识。候选人将获得小鼠和体外疾病建模的基本技能，并拓宽她的免疫学和上皮细胞学生物学知识库。这些技能将极大地促进候选人的职业发展独立的 NIH 资助的终身教职人员，其长期目标是了解控制粘膜修复和体内平衡的机制。