Keratinocyte Integrin Crosstalk During Wound Healing

伤口愈合过程中角质形成细胞整合素串扰

基本信息

批准号：
8623098
负责人：
C. Michael DiPersio
金额：
$ 33.58万
依托单位：
ALBANY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-02-20 至 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8623098
关键词：
Address Angiogenic Factor Basement membrane Biochemistry Bioinformatics Cell physiology Cells Cellular biology Chronic Cicatrix Communication Complex Custom Data Defect Deposition Diabetic ulcer Endothelial Cells Engineering Ensure Epidermis Extracellular Matrix Fibronectins Foundations Functional disorder Gene Expression Gene Expression Profile Gene Expression Regulation Genes Genetic Genomics Goals Growth Factor Healed Human Hypertrophic Cicatrix In Vitro Individual Injury Integrins Knock-out Knockout Mice Knowledge Laminin Ligands Link Mediating Molecular Molecular Profiling Mus Natural regeneration Outcome Paracrine Communication Peptides Play Process Publishing RNA Interference Rattus Recombinants Regulation Resolution Role Signal Transduction Staging Testing Therapeutic Time Work Wound Healing adhesion receptor angiogenesis base cDNA Arrays cell motility combinatorial healing in vivo keratinocyte laser capture microdissection meetings migration mouse model novel paracrine public health relevance receptor success therapeutic target wound

项目摘要

DESCRIPTION (provided by applicant): Epidermal keratinocytes are vital to normal wound healing by restoring the epidermal barrier and secreting paracrine factors that govern diverse processes including wound angiogenesis. In pathogenic settings, impaired epidermal function results in chronically insufficient (e.g., diabetic ulcers) or over-exuberant healing (e.g., hypertrophic scars). Our long-term goal is to develop therapeutic paradigms through which integrins can be manipulated to modulate pathogenic keratinocyte function. While it is well established that integrins regulate proliferation, migration and growth factor signaling, their rols in orchestrating wound keratinocyte functions remain enigmatic. Moreover, while normal and wound keratinocytes express integrin a9b1, in vivo, upon explanation integrin a9b1 is lost, confounding observations made in previous studies, in vitro. Using genetically defined, virally transduced keratinocytes that express integrins a3b1 and/or a9b1 in different combinations, we discovered that a9b1 exerts a cross-suppressive effect on cell functions and gene expression that is governed by a3b1, including paracrine signals that promote endothelial cell function. Moreover, we have derived genetically defined mice that express a3b1 and/or a9b1 in epidermis in different combinations. Strikingly, deletion of a9b1 from epidermis enhances wound contraction and angiogenesis, two functions that are attributed to paracrine signaling directed by a3b1. Based on our foundation data, we hypothesize that a9b1 suppresses a3b1-dependent paracrine signals from the epidermis that control wound closure and angiogenesis. We further hypothesize that the regulation of a9b1-mediated suppression of a3b1, perhaps through ligand-dependent activation of a9b1 at key stages of wound healing, is critical for proper temporal and spatial orchestration of epidermal functions that promote wound healing. This hypothesis will be tested in three Aims using a combination of genomics, bioinformatics, peptide biochemistry, cell biology, and defined genetic mouse models of wound healing. At the end of this project period, we will have provided the first analyses of keratinocyte a9b1 functions, determined how a3b1 and a9b1 coordinately regulate wound repair, identified molecular mechanisms through which a9b1 exerts cross- suppressive regulation over a3b1, and tested the concept that a9b1-targeting peptides can be used to control certain epidermal wound healing functions. In doing so, we will have developed the basis for novel integrin- targeting therapeutics to modulate keratinocyte functions and wound outcome.

描述（由申请人提供）：表皮角质形成细胞对正常伤口愈合至关重要，通过恢复表皮屏障和分泌旁分泌因子，这些旁分泌因子控制包括伤口血管生成在内的多种过程。在致病环境中，表皮功能受损会导致长期不足（例如糖尿病性溃疡）或过度过度愈合（例如，肥大性疤痕）。我们的长期目标是开发可以操纵整联蛋白以调节致病角质形成性功能的治疗范式。虽然整合蛋白可以调节增殖，迁移和生长因子信号传导，但它们在策划伤口角质形成细胞功能方面的ROL仍然神秘。此外，虽然正常和伤口的角质形成细胞在体内表达整联蛋白A9b1，但在解释中，整联蛋白A9B1丢失，但在先前的研究中进行了混杂的观察，体外。使用遗传定义的，病毒转导的角质形成细胞，这些角质形成细胞在不同组合中表达整联蛋白A3B1和/或A9B1，我们发现A9B1对细胞功能和基因表达产生了交叉抑制作用，对受A3B1的基因表达和基因表达效应，包括A3B1，包括促进内皮细胞功能的旁分泌信号。此外，我们衍生出在不同组合中表达表皮中表达A3B1和/或A9B1的基因定义的小鼠。令人惊讶的是，表皮中A9B1的缺失增强了伤口收缩和血管生成，这两个功能归因于A3B1指导的旁分泌信号。基于我们的基础数据，我们假设A9B1抑制了来自表皮的A3B1依赖性旁分泌信号，这些信号控制了伤口闭合和血管生成。我们进一步假设，通过在伤口愈合的关键阶段对A9B1的A9B1激活A9B1介导的A3B1抑制，对于促进伤口愈合的表皮功能的适当时间和空间编排至关重要。该假设将在三个目标中使用基因组学，生物信息学，肽生物化学，细胞生物学以及定义的伤口愈合遗传小鼠模型的组合进行检验。在该项目期间结束时，我们将提供角质形成细胞A9B1功能的首次分析，确定A3B1和A9B1如何协调调节伤口修复，确定了A9B1通过A3B1施加交叉抑制的分子机制，并测试了A9B1的概念 - 靶向肽可用于控制某些表皮伤口愈合功能。在此过程中，我们将为新型整合靶向治疗剂调节角质形成细胞功能和伤口预后的基础。