Keratinocyte Integrin Crosstalk During Wound Healing

伤口愈合过程中角质形成细胞整合素串扰

• 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
• 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

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）：表皮角质形成细胞通过恢复表皮屏障和分泌旁分泌因子对正常伤口愈合至关重要，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，受损的表皮功能导致慢性不足（例如，糖尿病溃疡）或过度旺盛的愈合（例如，增生性疤痕）。我们的长期目标是开发治疗范例，通过该治疗范例可以操纵整合素来调节致病性角质形成细胞功能。虽然整合素调节增殖、迁移和生长因子信号传导是公认的，但它们的整合素协调伤口角质形成细胞功能仍然是个谜。此外，虽然正常和创伤角质形成细胞在体内表达整联蛋白a9 b1，但在解释整联蛋白a9 b1丢失时，在先前的体外研究中进行了混淆观察。使用基因定义的，病毒转导的角质形成细胞，表达整合素a3 b1和/或a9 b1在不同的组合，我们发现，a9 b1发挥交叉抑制作用的细胞功能和基因表达，是由a3 b1，包括旁分泌信号，促进内皮细胞功能。此外，我们已经衍生出在表皮中以不同组合表达a3 b1和/或a9 b1的遗传定义的小鼠。引人注目的是，从表皮中删除a9 b1增强了伤口收缩和血管生成，这两种功能归因于a3 b1指导的旁分泌信号传导。基于我们的基础数据，我们假设a9 b1抑制a3 b1依赖的旁分泌信号从表皮控制伤口闭合和血管生成。我们进一步假设，调节a9 b1介导的抑制a3 b1，也许通过在伤口愈合的关键阶段的配体依赖性激活a9 b1，是至关重要的适当的时间和空间编排表皮功能，促进伤口愈合。将使用基因组学、生物信息学、肽生物化学、细胞生物学和定义的伤口愈合遗传小鼠模型的组合，在三个目标中测试该假设。在该项目期结束时，我们将提供角质形成细胞a9 b1功能的第一次分析，确定a3 b1和a9 b1如何协调调节伤口修复，鉴定a9 b1对a3 b1发挥交叉抑制调节的分子机制，并测试a9 b1靶向肽可用于控制某些表皮伤口愈合功能的概念。在这样做的过程中，我们将为新型整联蛋白靶向治疗剂开发基础，以调节角质形成细胞功能和伤口结果。