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

• 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
• 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

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)排列在整个肠道内表面，处理所有 体内营养和液体的吸收，同时起到阻挡有毒管腔的作用 内容物和病原体。对抗人类肠道上皮细胞(HIECs)的损伤 在动态平衡期间积累，绒毛(初级吸收表面)上的细胞不断 被替换了。隐窝中的专用肠道干细胞(HISCs)持续增殖，以产生 源源不断的祖细胞集体向绒毛迁移，是最 成人体内集体细胞迁移的扩张性例子。在IEC受伤或丢失后， 附近的IEC经历了一种名为恢复原状的快速集体迁移，以迅速修复损坏。 由集体移徙推动的恢复是保护这一障碍的一个基本的细胞过程 反对流明的内容。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。与WNT相关的平面细胞的极性 (WPCP)途径，特别是细胞表面蛋白Vangl2，已经越来越多地 被认为是协调集体细胞迁移的主要参与者，但研究 由于用于研究WPCP的生理相关模型系统的主要缺陷 高等有机体。从我的初步数据中观察到：1)零星的 Vangl2的过度表达足以强烈地损害集体迁移，2)Vangl2是 在划痕试验中在上皮伤口的前缘上调，以及3)蛋白质梯度 在体内沿隐窝-绒毛轴分布。综合这些观察结果，我得出了 假设IE利用WPCP途径来协调集体细胞迁移。 我将用两个目标来检验这一假设。目标1将为WPCP在#年建立一个机械作用 动态平衡迁移，而Aim 2将确定其在伤口愈合中的作用。这两个目标都将使用 WPCP途径扰动的新的可诱导遗传模型以产生原始数据。这 将被用来训练WPCP极性的计算反应-扩散模型，以破译 局部WPCP反馈可以如此强烈地影响大组织区域的集体迁移。这 研究将通过生成新的文化模型来填补关键的技术和知识空白 人类细胞，揭示了HIE如何控制集体迁移的新机制，以及 为研究集体迁徙和WPCP建立新的计算模型。我的发现将 Vangl2和WPCP配体与多种健康相关的重要性 包括发育缺陷、神经和肾脏疾病、骨骼肌和 胆管再生、癌症转移和慢性结肠炎。