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

sEV 相关 miR-146a 在组织修复中的作用

• 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
• 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

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 皮肤伤口代表了一个重大的医疗保健问题，自1997年以来，没有治疗慢性伤口 已获得FDA批准。为这种治疗空间做出贡献是伤口愈合过程的复杂性 相对理解的领域，这些活动如何在慢性伤口中失调和利用 和细胞转化。我们以前的工作表明钙粘蛋白Desmoglein 2（DSG2）是一个 在肿瘤发生和 伤口愈合。我们已经表明，DSG2在慢性伤口中被下调，并且本构成 表皮中DSG2的过表达供应增加伤口愈合，过度增殖和肿瘤 发展。强调改变角质形成细胞动力学对许多病理学的贡献 皮肤病疾病，包括慢性伤口，牛皮癣和鳞状细胞癌。 DSG2能够 调节角质形成活性，例如增殖和通过其下游调节的迁移 信号分子包括促生蛋白和迁移蛋白：EGFR和C-SRC。除了 蛋白质的调节，我们的实验室还证明DSG2调节细胞外和小细胞外 囊泡（SEV）转录组。 mirnaseq揭示了DSG2诱导miR-146a的巨大耗竭，该耗竭 是炎症反应的潜在调节剂，已被证明可能调节4469个基因，即 包括miR-155的水平。因此，有趣的是注意mir-155在 对DSG2表达的响应。最近，我们专注于确定DSG2的上游监管机构，以及 令我们惊讶的是，miR-155的目标是DSG2，这应该导致其降解，但是，这些dsg2的rnaseq是 相同的细胞导致人们意识到lncRNA的水平也增加了DSG2-AS1。很好 确定lncrnas可以“海绵” miRNA并抑制其靶标的降解。这导致了我们 考虑一种新型的调节机制，DSG2-AS1海绵miR-155，可以显着增加 在DSG2表达中。初步数据表明，这种机制在正常伤口愈合期间正在发挥作用， 如MiR-155和DSG2-AS1同时增加后，DSG2表达的增加所证明 在伤口修复期间。在此提案中，我们假设控制DSG2表达的调节循环是 慢性伤口（例如糖尿病的背景下）改变了，这导致了角质形成细胞激活 因此无效的伤口闭合。这是该提议的目标是阐明DSG2监管 网络及其对角质形成细胞增殖和迁移的影响，并证明了SEV的能力 封装靶向该网络的RNA物种是刺激伤口修复中角质形成细胞激活的一种方式。 从这些研究中获得的发现将证明SEV作为强大的治疗方式的可行性 并有助于对伤口愈合的机械理解。