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

• 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
• 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

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.

项目摘要 炎症性肠病（IBD）是一系列不可治愈的、复发性和缓解性疾病， 肠粘膜屏障功能缺陷和异常的促炎反应， 微生物植物群。IBD患者发生结肠炎相关结肠直肠癌（CAC）的风险增加， 控制它的上皮特异性机制还不完全清楚。同样不清楚的是， 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 揭示了一种新的上皮修复蛋白，富含亮氨酸的重复序列和含有IG结构域， Nogo受体相互作用蛋白家族2（LINGO2）有助于恶性转化。初步 数据，我们表明上皮细胞中的Lingo2缺陷导致EGFR磷酸化的上调， 与野生型相比，响应于IL 6处理的STAT3活化延长并且增殖增加 细胞此外，Lingo2缺陷型小鼠具有严重的葡聚糖硫酸钠（DSS）诱导的结肠炎， 通过EGFR抑制来拯救。我们证明了Lingo2在免疫细胞中表达，并且在免疫细胞中的损失。 造血区室有助于结肠炎的易感性。与发表的观察结果一致 我们发现，增加的EGFR信号传导具有促肿瘤作用，当暴露于氧化偶氮甲烷（AOM）和 与对照小鼠相比，DSS，Lingo2缺陷型小鼠更容易发生CAC。我 因此，假设Lingo2通过调节EGFR防止CAC的发展， 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的保护，然后了解微环境中的Lingo2缺乏是否是 对CAC来说很重要我的假设将通过两个相互关联的具体目标来检验， CAC中上皮细胞中LINGO2和EGFR信号传导的相互作用（Aim 1），然后评估Lingo2 缺乏影响CAC中STAT3活化细胞因子的产生和骨髓细胞的活性（目的2）。 所提出的实验将使用各种创新的方法，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 因为它将提供关于新的肿瘤抑制分子途径的新信息。 宾夕法尼亚大学提供了完美的研究环境，进行这项调查，因为当地 在粘膜生物学、上皮生物学和免疫学的相关领域的专业知识。候选人将 获得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 生物学知识库这些技能组合将大大提高候选人的职业发展， 独立的NIH资助的终身教职员工的长期目标是了解 控制粘膜修复和体内平衡的机制。