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.

抽象的。肠上皮（IE）在肠道的整个内部表面上排成一条，处理所有人体的营养和液体吸收，同时充当有毒腔的障碍内容和病原体。打击损害人类肠上皮细胞（HIEC）在体内稳态期间积聚，绒毛上的细胞（主要吸收表面）一直是更换。隐窝中的专用肠干细胞（HISC）连续增殖以产生祖细胞的稳定流，这些细胞集体朝着最多的绒毛迁移到绒毛成人体内集体细胞迁移的广泛例子。在IEC受伤或损失之后，附近的IEC经历了一个快速的集体迁移，称为修复，以快速修复损坏。集体迁移驱动的恢复原状是保留障碍的必不可少的细胞过程针对腔内内容。尽管集体细胞迁移到肠功能很重要，但关于HIEC如何协调这一过程，知之甚少。 Wnt相关的平面细胞极性（WPCP）途径，尤其是细胞表面蛋白vangl2，越来越多作为集体细胞迁移协调的主要参与者实施，但研究是由于对研究WPCP的物理相关模型系统的重大缺陷而受到阻碍更高的生物。从我的初步数据中观察到：1）零星 vangl2的过表达足以严重损害集体迁移，2）vangl2是在划痕的上皮伤口的前缘上调，3）蛋白质梯度 vangl2的沿体内加密villus轴存在。这些观察结果使我假设IE利用WPCP途径来协调集体细胞迁移。我将使用两个目标检验这一假设。 AIM 1将在WPCP中建立机械作用稳态迁移，而AIM 2将确定其在伤口愈合中的作用。两个目标都将使用 WPCP途径扰动的新型诱导遗传模型生成主要数据。这将用于训练WPCP极性的计算反应扩散模型，以破译局部WPCP反馈会严重影响大型组织区域的集体迁移。这研究将通过产生新的文化模型来填补关键的技术和知识差距人类细胞，发现HIE如何控制集体迁移的新机制，构建用于研究集体迁移和WPCP的新型计算模型。我的发现会随着vangl2和WPCP配体越来越多地与潜水员健康有关包括发育缺陷，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。