Investigating a role for Wnt-associated planar polarity in collective migration of human intestinal epithelium

研究 Wnt 相关平面极性在人肠上皮集体迁移中的作用

基本信息

批准号：
10751956
负责人：
Keith Breau
金额：
$ 3.91万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10751956
关键词：
Adopted Adult Affect Applications Grants Biliary Biological Models Cell Polarity Cell Surface Proteins Cell Surface Receptors Cell physiology Cell-Cell Adhesion Cells Chronic Colitis Complex Computer Models Confocal Microscopy Data Dedications Defect Development Devices Differential Equation Diffusion Disease Distal Drosophila genus Duct (organ) structure Epithelial Cells Epithelium Feedback Formulation Genetic Models Goals Grant Health Homeostasis Human Hybrids Impairment Individual Injury Intercellular Junctions Intervention Intestines Kidney Diseases Knock-out Knowledge Ligands Liquid substance Machine Learning Measures Membrane Mentorship Methods Modeling Natural regeneration Neoplasm Metastasis Nutrient Organism Paper Pathway interactions Phenotype Physiological Play Process Proliferating Proteins Publishing Reaction Regulation Research Role Side Signal Transduction Skeletal Muscle Specific qualifier value Speed Stains Stream Study models Surface Testing Therapeutic Intervention Tissues Training Villus Writing biological systems cell injury cell motility combat design epithelial wound fabrication human disease human migration human model imaging Segmentation in vivo injury and repair intestinal epithelium knock-down mathematical model migration monolayer mutant nervous system disorder novel overexpression pathogen physiologic model planar cell polarity preservation repaired small hairpin RNA spatiotemporal stem cells tool treatment strategy uptake virtual wound wound healing

项目摘要

Abstract. The intestinal epithelium (IE) lines the entire inner surface of the intestine, handling all of the body’s nutrient and fluid uptake while simultaneously serving as a barrier to toxic luminal contents and pathogens. To combat the damage human intestinal epithelial cells (hIECs) accumulate during homeostasis, cells on the villi (primary absorptive surfaces) are constantly replaced. Dedicated intestinal stem cells (hISCs) in crypts proliferate continuously to generate a steady stream of progenitor cells that collectively migrate towards the villi in one of the most expansive examples of collective cell migration in the adult body. Following IEC injury or loss, nearby IECs undergo a rapid collective migration called restitution to quickly repair the damage. Restitution driven by collective migration is an essential cellular process to preserve the barrier against luminal contents. Despite the importance of collective cell migration to intestinal function, little is known about how hIECs coordinate this process. The WNT-associatedPlanar Cell Polarity (WPCP) pathway, and in particular the cell surface protein VANGL2, has been increasingly implicated as a major player in the coordination of collective cell migration, but research is hampered due to a major deficit of physiologically relevant model systems for studying WPCP in higher organisms. Observations from my preliminary data indicate that: 1) sporadic overexpression of VANGL2 is sufficient to strongly impair collective migration, 2) VANGL2 is upregulated at the leading edge of an epithelial wound in scratchassays, and 3) a protein gradient of VANGL2 exists along the in vivo crypt-villus axis. Together these observations led me to hypothesize that the IE utilizes the WPCP pathway to coordinate collective cell migration. I will test this hypothesis using two aims. Aim 1 will establish a mechanistic role for WPCP in homeostatic migration, while Aim 2 will establish its role in wound healing. Both aims will use novel inducible genetic models of WPCP pathway perturbations to generate primary data. This will be used to train a computational reaction-diffusion model of WPCP polarity to decipher how local WPCP feedback can so strongly impact collective migration of large tissue regions. This study will fill critical technical and knowledge gaps by generating new culture models with primary human cells, uncovering novel mechanisms of how the hIE controls collective migration, and building novel computational models for studying collective migration and WPCP. My findings will be significant as VANGL2 and WPCP ligands are increasingly implicated in diverse health conditions including developmental defects, neurological and kidney disease, skeletal muscle and biliary duct regeneration, cancer metastasis, and chronic colitis.

摘要：肠上皮 (IE) 排列在肠道的整个内表面，负责处理所有的事情。身体的营养和液体吸收，同时作为有毒管腔的屏障对抗损伤的人肠上皮细胞（hIEC）。在体内平衡过程中积累，绒毛（主要吸收表面）上的细胞不断地隐窝中的专用肠道干细胞（hISC）不断增殖以产生祖细胞源源不断地集体向绒毛迁移，这是最常见的细胞之一 IEC 损伤或丧失后，成人体内集体细胞迁移的例子不断增多。附近的 IEC 会进行快速集体迁移（称为恢复原状），以快速修复损坏。由集体迁移驱动的恢复是保护屏障的重要细胞过程尽管集体细胞迁移对肠道功能很重要，关于 hIEC 如何协调这一过程，人们知之甚少。 (WPCP) 途径，特别是细胞表面蛋白 VANGL2，已越来越多地被被认为是协调集体细胞迁移的主要参与者，但研究表明由于研究 WPCP 的生理相关模型系统的重大缺陷而受到阻碍我的初步数据观察表明：1）零星的。 VANGL2 的过度表达足以严重损害集体迁移，2) VANGL2 是在划痕试验中上皮伤口前缘上调，3) 蛋白质梯度 VANGL2 存在于体内隐窝-绒毛轴上，这些观察使我得出结论。 IE 利用 WPCP 途径来协调集体细胞迁移。我将使用两个目标来检验这一假设：目标 1 将在 WPCP 中发挥机械作用。稳态迁移，而目标 2 将确定其在伤口愈合中的作用。 WPCP 通路扰动的新型诱导遗传模型可生成原始数据。将用于训练 WPCP 极性的计算反应扩散模型，以破译如何局部 WPCP 反馈可以强烈影响大组织区域的集体迁移。研究将通过产生新的文化模型来填补关键的技术和知识空白人类细胞，揭示 hIE 如何控制集体迁移的新机制，以及我的研究结果将建立新的计算模型来研究集体迁移和 WPCP。意义重大，因为 VANGL2 和 WPCP 配体越来越多地涉及多种健康疾病包括发育缺陷、神经系统和肾脏疾病、骨骼肌和胆管再生、癌症转移和慢性结肠炎。