Characterization of Keloid Specific Exosomes and Determination of Exosomal Critical Signaling Pathways in the Keloid Microenvironment

瘢痕疙瘩特异性外泌体的表征以及瘢痕疙瘩微环境中外泌体关键信号通路的测定

• 批准号: 10443732
• 负责人: Lamont Jones
• 金额: $ 17.93万
• 依托单位: HENRY FORD HEALTH SYSTEM
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10443732
• 关键词: Affect; Animal Model; Benign; Biological Markers; Cell Communication; Cell Line; Cell physiology; Cells; Cicatrix; Clinic Visits; Data; Development Plans; Disease; Ear; Ethnic group; Event; Exercise; Fibroblasts; Fibrosis; Genes; Goals; Investigation; Keloid; Mediating; Mentors; Methylation; MicroRNAs; Modeling; Oryctolagus cuniculus; Outcome; Pain; Pathogenesis; Pathway interactions; Patients; Persons; Phenotype; Physiological; Positioning Attribute; Production; Pruritus; Publishing; Recurrence; Research; Research Proposals; Resources; Role; Scientist; Signal Pathway; Signal Transduction; Skin; Skin Tissue; Testing; Therapeutic; Treatment outcome; United States; Work; base; career development; emotional distress; epigenetic marker; exosome; improved; improved outcome; in vivo; innovation; insight; intercellular communication; loss of function; multi-ethnic; novel; success; targeted treatment; translational applications; tumor; wound healing

Project Summary/Abstract: There are more than 11 million people in the world with keloids and more than 425,000 associated clinic visits, yearly, in the United States. Keloids are benign fibroproliferative tumors which cause pain, pruritus, emotional distress and loss of function. Current therapies are unsatisfactory with unacceptably high recurrence rates, mainly because of an incomplete understanding of keloid pathogenesis. Fibroblasts are a key player in keloid pathogenesis, but the drivers are unknown. Keloid disease is influenced by aberrant signaling pathways. However, no clear signaling pathway has been identified. Exosomes mediate cell-cell communication, exercising primary physiological and pathophysiological function. Exosomal cargo, such as microRNAs (miRNAs), regulate cellular function. Our group identified RAB27, important for exosome secretion, as being differentially hypomethylated in keloid compared to normal skin. Our group has isolated keloid-specific exosomes. To date, there are no published studies on keloid exosomes or the contribution of RAB27 methylation on exosome function. We propose to test the central hypothesis that exosomes communicate critical signaling events in the keloid microenvironment mediated by RAB27 gene methylation. Aim 1: To determine the effect of RAB27 methylation on keloid exosome production and miRNA cargo profiles. Hypotheses: (1) Keloid exosomal production correlates with RAB27 gene methylation. (2) Keloid exosomal cargo miRNA expression profiles correlates with RAB27 gene methylation. (3) Keloid exosome miRNA’s putative target genes lie within pathways essential for wound healing and/or fibrosis. Aim 2: To determine the effects of keloid exosomes on the keloid microenvironment. Hypothesis: Keloid fibroblast exosomes compared to normal fibroblast exosomes will cause pro-fibrotic phenotype changes in normal fibroblasts. Aim 3: To determine the effect of keloid exosomes on scar formation in vivo. Hypothesis: Exosomes generated in aim 1 and tested in aim 2 will increase scar formation in a rabbit ear scar model. Significance: This project will lead to an enhanced understanding of keloid pathogenesis and the potential for exosome-based therapy. Innovation: (i) rational progression from preliminary data supporting the novel role of exosomes in keloid pathogenesis; (ii) investigating the influence of RAB27 methylation on the function and production of keloid exosomes would suggest a mechanistic basis for novel epigenetic biomarkers;(iii) using unique resources which includes fibroblast cell lines from primary untreated keloid (25) and matched normal skin (25) from a multi-ethnic group of patients and an in vivo animal model allow for the pragmatic translational application of results; (iv) entirely new field of keloid investigation. In summary, this project, mentoring and career development plan will position, Lamont R Jones, MD, MBA, to become an independent clinician scientist and leader in keloid pathogenesis.

项目概要/摘要：世界上有超过1100万人患有瘢痕疙瘩及以上 在美国，每年有超过425，000次相关的诊所访问。瘢痕疙瘩是良性纤维增生性的 引起疼痛、瘙痒、情绪困扰和功能丧失的肿瘤。目前的治疗方法是 不能令人满意，复发率高得令人无法接受，主要是因为对 瘢痕疙瘩的发病机制成纤维细胞是瘢痕疙瘩发病机制中的关键角色，但驱动因素尚不清楚。 瘢痕疙瘩疾病受异常信号通路的影响。然而，没有明确的信号通路 被识别。外泌体介导细胞-细胞通讯，行使主要的生理和 病理生理功能外泌体货物，如microRNA（miRNA），调节细胞功能。 我们的研究小组鉴定了对外泌体分泌很重要的RAB 27，它在细胞中被差异低甲基化。 与正常皮肤相比，我们的小组已经分离出瘢痕疙瘩特异性外泌体。迄今为止，没有 已发表的关于瘢痕疙瘩外泌体的研究或RAB 27甲基化对外泌体功能的贡献。 我们建议测试中心假设，即外泌体在细胞内传递关键信号事件。 瘢痕疙瘩微环境介导的RAB 27基因甲基化目的1：确定 RAB 27甲基化对瘢痕疙瘩外泌体产生和miRNA货物谱的影响假设：（1）瘢痕疙瘩 外泌体产生与RAB 27基因甲基化相关。(2)瘢痕疙瘩外泌体货物miRNA 表达谱与RAB 27基因甲基化相关。(3)瘢痕疙瘩外泌体miRNA的假定靶点 基因位于伤口愈合和/或纤维化所必需的途径内。目标2：确定影响 瘢痕疙瘩外泌体对瘢痕疙瘩微环境的影响假设：瘢痕疙瘩成纤维细胞外泌体比较 与正常成纤维细胞外泌体结合将导致正常成纤维细胞中的促纤维化表型变化。目标三： 确定瘢痕疙瘩外泌体对体内瘢痕形成的影响。假设：外泌体 在AIM 1中产生并在AIM 2中测试的化合物将增加兔耳瘢痕模型中的瘢痕形成。 意义：该项目将导致对瘢痕疙瘩发病机制的深入了解， 基于外泌体的治疗的潜力。创新：（一）根据初步数据进行合理的进展 支持外来体在瘢痕疙瘩发病机制中的新作用;（ii）研究RAB 27对瘢痕疙瘩的影响， 甲基化对瘢痕疙瘩外泌体的功能和产生的影响将表明瘢痕疙瘩外泌体甲基化的机制基础。 新的表观遗传生物标志物;（iii）使用独特的资源，包括来自原代的成纤维细胞系; 未治疗的瘢痕疙瘩（25例）和匹配的正常皮肤（25例），来自多种族患者组， 动物模型允许结果的实际转化应用;（iv）瘢痕疙瘩的全新领域 调查总之，本项目，指导和职业发展计划将定位，拉蒙R 琼斯，医学博士，工商管理硕士，成为一个独立的临床科学家和领导人在瘢痕疙瘩发病机制。

项目成果

MicroRNA Profile Differentiates Head and Neck Keloid and Adjacent Normal Skin Tissue.

MicroRNA 谱可区分头颈部瘢痕疙瘩和邻近的正常皮肤组织。

• DOI: 10.1089/fpsam.2020.0414
• 发表时间: 2021
• 期刊: Facial plastic surgery & aesthetic medicine
• 影响因子: 2
• 作者: Jones,LamontR;Levin,AlbertM;Dai,Xiangguo;Datta,Indrani;Li,Jia;Yin,Congcong;Mi,Qing-Sheng
• 通讯作者: Mi,Qing-Sheng