Wound healing mechanisms modulated by novel antimicrobial Peptides

新型抗菌肽调节伤口愈合机制

• 批准号: 9182311
• 负责人: Anne Kasus-Jacobi
• 金额: $ 18.5万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-30 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9182311
• 关键词: Adhesions; Antibiotic Resistance; Antibiotics; Antimicrobial Effect; Bacteria; Bacterial Infections; Binding; Biological Process; Blindness; Cells; Clinic; Clinical; Co-Immunoprecipitations; Complex; Cornea; Corneal Abrasion; Corneal Injury; Dermal; Development; Disease; Ensure; Epithelial Cells; Event; Eye Injuries; G-Protein-Coupled Receptors; Goals; Healed; Health; Human; Immune system; Infection; Inflammation; Intracellular Membranes; Intracellular Signaling Proteins; Invaded; Investigation; Keratitis; Knowledge; Lead; Length; Ligand Binding; Ligands; Mediating; Mediator of activation protein; Membrane; Methods; Molecular; Mus; National Eye Institute; One-Step dentin bonding system; Peptides; Prevention; Process; Protein Analysis; Protein Isoforms; Proteins; RNA analysis; Reporting; Research; Resistance; Risk Factors; Role; Signal Pathway; Signal Transduction; Structure; Tertiary Protein Structure; Testing; Therapeutic; Time; Translating; United States; Vision; Work; Wound Healing; analog; antimicrobial; antimicrobial peptide; base; cell motility; clinical application; corneal epithelium; extracellular; healing; human migration; improved outcome; in vivo; inhibitor/antagonist; innovation; killings; knock-down; migration; novel; novel therapeutics; open wound; pathogen; peptide analog; peptide drug; prevent; receptor; receptor binding; research study; response; wound

DESCRIPTION: Wound healing is a complex process that follows a defined sequence of events consisting of inflammation, proliferation and migration of epithelial cells to re-epithelialize the wound, and remodeling/differentiation. The cationic antimicrobial protein of 37 kDa (CAP37) belongs to the innate immune system and was originally identified based on its powerful antimicrobial effect. It was later found to induce migration, adhesion and proliferation of cultured corneal epithelial cells, which translated into faster corneal re- epithelialization in vivo. Structure-function studies of CAP37 were initiated with the long-term goal to identify the antimicrobial and the wound healing domains of the protein to generate therapeutic peptides for clinical use. Our lab recently identified two CAP37-derived peptide analogs that carry a strong antimicrobial activity, even on antibiotic resistant or multiresistant strains, and a wound healing activity. These analogs have strong potential to be used as innovative ocular treatments with the dual effects of promoting corneal re- epithelialization and preventing or clearing bacterial infections. The molecular mechanisms mediating the wound healing effects of the full length CAP37 and the two derived analogs are unknown. The objective of this proposal is to identify the membrane and intracellular mediators activated by these extracellular ligands to induce migration of immortalized corneal epithelial cells and corneal re-epithelialization in mouse. Based on previous studies in our lab, we hypothesize that CAP37 and derived analogs act as extracellular ligands, binding a G protein-coupled receptor (GPCR) on corneal epithelial cells to activate intracellular signaling pathways, including the signal transduction proteins PKCδ and PKCθ, thus promoting wound healing effects. Two specific aims are proposed. (1) To identify the receptor(s) mediating the migration of human corneal epithelial cells and corneal wound healing in response to CAP37 ligands. (2) To identify the downstream signaling pathway(s) mediating corneal epithelial cell migration and corneal wound healing in response to CAP37 ligands. This new knowledge will break new grounds in this field of research because a receptor for CAP37 will be identified for the first time since its discovery 30 years ago. This new knowledge will also be key to developing and optimizing CAP37-derived peptide analogs into innovative therapeutics with the dual effects of killing resistant pathogens and promoting wound healing.

描述：伤口愈合是一个复杂的过程，遵循由以下事件组成的定义序列： 炎症、上皮细胞增殖和迁移以使伤口再上皮化，和 重塑/分化。37 kDa的阳离子抗菌蛋白（CAP 37）属于先天性免疫 系统，最初是根据其强大的抗菌作用确定的。后来发现它能诱导 培养的角膜上皮细胞的迁移、粘附和增殖，从而转化为更快的角膜重建。 体内上皮形成。启动了CAP 37的结构-功能研究，其长期目标是鉴定 所述蛋白质的抗微生物和伤口愈合结构域以产生用于临床的治疗肽 使用.我们的实验室最近鉴定了两种CAP 37衍生的肽类似物，它们具有很强的抗微生物活性， 甚至对抗生素抗性或多重抗性菌株，以及伤口愈合活性。这些类似物具有很强的 有可能作为创新的眼部治疗方法，具有促进角膜再生的双重效果， 上皮形成和预防或清除细菌感染。 介导全长CAP 37和两个衍生的CAP 37的伤口愈合作用的分子机制 类似物未知。本提案的目的是确定膜和细胞内介质 由这些细胞外配体激活以诱导永生化角膜上皮细胞和角膜上皮细胞的迁移， 再上皮化。基于我们实验室以前的研究，我们假设CAP 37和衍生的 类似物作为细胞外配体，结合角膜上皮细胞上的G蛋白偶联受体（GPCR）， 激活细胞内信号通路，包括信号转导蛋白PKCδ和PKCθ，从而 促进伤口愈合效果。提出了两个具体目标。(1)为了鉴定介导细胞凋亡的受体 人角膜上皮细胞的迁移和角膜创伤愈合对CAP 37配体的应答。(2)到 鉴定介导角膜上皮细胞迁移和角膜创伤的下游信号通路 对CAP 37配体的应答愈合。这一新知识将在这一研究领域开辟新天地 因为这将是自30年前发现CAP 37以来首次发现CAP 37的受体。这个新 知识也将是开发和优化CAP 37衍生肽类似物的关键， 具有杀灭耐药病原体和促进伤口愈合双重效果的治疗剂。