The Role of sEV-associated miR-146a in Tissue Repair

sEV 相关 miR-146a 在组织修复中的作用

基本信息

批准号：
10538174
负责人：
Brianna Leigh Hill
金额：
$ 4.68万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-27 至 2024-09-26
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10538174
关键词：
Acceleration Biological Biological Assay Cadherins Cell Line Cells Clinical Trials Data Dermatologic Development Diabetes Mellitus Diabetic mouse Encapsulated Engineering Environment Epidermal Growth Factor Receptor Epidermis Event Expenditure Genes Goals Healthcare Hyperplasia Inflammatory Response Lead Malignant - descriptor Malignant Epithelial Cell Mediating Medicare Messenger RNA MicroRNAs Modality Nature Outcome Pathology Pathway interactions Play Porifera Process Proteins Psoriasis RNA Regulation Research Role SRC gene Signal Transduction Signaling Molecule Squamous cell carcinoma Subcutaneous Injections Supplementation System Up-Regulation Vesicle Western Blotting Work Wound healing therapy chronic wound desmoglein 2 diabetic wound healing experimental study extracellular vesicles healing inhibitor interest keratinocyte metaplastic cell transformation migration mouse model novel overexpression prevent response skin wound tissue regeneration tissue repair transcriptome transcriptome sequencing tumor tumorigenesis wound wound closure wound healing wound treatment

项目摘要

ABSTRACT Cutaneous wounds represent a significant healthcare issue and since 1997, no treatment for chronic wounds has received FDA approval. Contributing to this treatment desert is the complexity of the wound healing process and the relatively understudied field of how these activities are dysregulated and exploited in chronic wounds and cellular transformation. Our previous work demonstrated that the cadherin, desmoglein 2 (Dsg2) is a pleiotropic protein that modulates mitogenic signaling and pro-migratory programming in both tumorigenesis and wound healing. We have shown that Dsg2 is down-regulated in chronic wounds and that constitutive overexpression of Dsg2 in the epidermis confers increased wound healing, hyperproliferation, and tumor development. Highlighting the contribution of altered keratinocyte dynamics to the pathology of many dermatological conditions including chronic wounds, psoriasis, and squamous cell carcinoma. Dsg2 is able to modulate keratinocyte activities such as proliferation and migration through its regulation of downstream signaling molecules including the pro-proliferative and migratory proteins: EGFR and c-Src. In addition to the modulation of proteins, our lab has also demonstrated that Dsg2 regulates the cellular and small extracellular vesicle (sEV) transcriptome. miRNAseq revealed that Dsg2 induced the dramatic depletion of miR-146a, which is a potent regulator of the inflammatory response and has been shown to potentially regulate 4469 genes, including the level of miR-155. Therefore, it was interesting to note the consequent increase of miR-155 in response to Dsg2 expression. Recently, we have focused on identifying the upstream regulators of Dsg2, and much to our surprise, miR-155 targets Dsg2, which should lead to its degradation, however, RNAseq of these same cells lead to the realization that the level of the lncRNA, Dsg2-AS1, was also increased. It is well established that lncRNAs can “sponge” miRNAs and inhibit the degradation of their targets. This led us to consider a novel regulatory mechanism whereby Dsg2-AS1 sponges miR-155, allowing for a dramatic increase in Dsg2 expression. Preliminary data demonstrates that this mechanism is at play during normal wound healing, as evidenced by the increase in Dsg2 expression after the concomitant increase of miR-155 and Dsg2-AS1 during wound repair. In this proposal, we hypothesize that the regulatory loop that controls Dsg2 expression is altered in chronic wounds such as in the context of diabetes, which results in perturbed keratinocyte activation and therefore ineffective wound closure. Thus, the goals of this proposal are to elucidate the Dsg2 regulatory network and its impact on keratinocyte proliferation and migration, and to demonstrate the ability of sEV- encapsulated RNA species that target this network as a way to stimulate keratinocyte activation in wound repair. The findings obtained from these studies will demonstrate the feasibility of sEVs as a robust treatment modality and contribute to the mechanistic understanding of wound healing.

抽象的皮肤伤口是一个重要的医疗保健问题，自 1997 年以来，慢性伤口没有治疗方法已获得 FDA 批准的原因是伤口愈合过程的复杂性。以及这些活动如何在慢性伤口中失调和利用的研究相对较少的领域我们之前的工作表明，钙粘蛋白、桥粒芯糖蛋白 2 (Dsg2) 是一种调节肿瘤发生和迁移过程中的有丝分裂信号传导和促迁移编程的多效性蛋白我们已经证明 Dsg2 在慢性伤口中表达下调，并且持续存在。 Dsg2 在表皮中的过度表达可促进伤口愈合、过度增殖和肿瘤生长强调角质形成细胞动力学改变对许多病理学的贡献。 Dsg2 能够治疗慢性伤口、牛皮癣和鳞状细胞癌等皮肤病。通过调节下游的角质形成细胞活性，例如增殖和迁移信号分子包括促增殖和迁移蛋白：EGFR 和 c-Src。蛋白质的调节，我们的实验室还证明 Dsg2 调节细胞和小细胞外囊泡 (sEV) 转录组显示 Dsg2 诱导 miR-146a 的急剧消耗，这是炎症反应的有效调节剂，已被证明可以调节 4469 个基因，包括 miR-155 的水平，因此，值得注意的是 miR-155 随之增加。最近，我们重点鉴定了 Dsg2 的上游调控因子，以及令我们惊讶的是，miR-155 靶向 Dsg2，这应该会导致其降解，然而，这些的 RNAseq 相同的细胞导致认识到 lncRNA、Dsg2-AS1 的水平也有所增加。确定 lncRNA 可以“海绵”miRNA 并抑制其靶标的降解。考虑一种新的调节机制，通过 Dsg2-AS1 海绵 miR-155，允许急剧增加初步数据表明，这种机制在正常伤口愈合过程中发挥作用， miR-155 和 Dsg2-AS1 同时增加后 Dsg2 表达增加证明了这一点在伤口修复过程中，我们发现控制 Dsg2 表达的调节环是慢性伤口（例如糖尿病）发生改变，导致角质形成细胞活化紊乱因此，该提案的目标是阐明 Dsg2 监管。网络及其对角质形成细胞增殖和迁移的影响，并证明 sEV- 封装的RNA种类以该网络为目标，作为刺激伤口修复中角质形成细胞活化的一种方式。这些研究的结果将证明 sEV 作为一种稳健的治疗方式的可行性并有助于对伤口愈合机制的理解。