TARGETING MIR-29 FUNCTION IN CUTANEOUS WOUND REPAIR

针对皮肤伤口修复中的 MIR-29 功能

• 批准号: MR/P009700/1
• 负责人: Svitlana Kurinna
• 金额: $ 153.92万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FP009700%2F1
• 关键词: TARGETING; MIR; 29; FUNCTION; CUTANEOUS

The incidence of non-healing skin ulcers and the need of improved skin regeneration is continuously increasing in our society. For the development of efficient strategies to improve skin repair it is essential to understand the mechanisms underlying normal and impaired healing. For my project, I am seeking to further describe functions of small ribonucleic acids (RNA) molecules, miR-29s, which I identified as regulators of the top layer of the skin called the epidermis. Upon wounding, the cells of the epidermis (mostly keratinocytes) are responsible for covering the wound site with a new layer of skin that protects our body from infection entry and water loss. Keratinocytes can sufficiently grow and migrate into the wound site only in a healthy skin. It is very important to understand how this process is regulated in the normal skin, and then how this regeneration of keratinocytes is deregulated in patients with impaired wound healing. The overall goal of this research is to enhance normal growth of keratinocytes and thereby, regeneration of the epidermis. However, on the other hand, the excessive growth of keratinocytes results in a hyper-thickened skin, cracking of the epidermis, water loss, and severe secondary infections. Thus, it is important to find molecules that regulate and ensure a fine-tuned control over the normal growth of keratinocytes. Small RNAs, like miR-29s, have been already described to function as precise and physiologic regulators of normal skin growth. Importantly, because of the small size and chemical properties of short nucleic acids, miRs can be used for molecular therapy. miR-29s have already been tried in the clinic to improve skin condition in patients with cutaneous sclerosis (manifested by the abnormal skin thickening), and thus can potentially be rapidly used for molecular therapy of other skin diseases, including wounds. However, it requires knowing all possible molecules that interact with miR-29s inside keratinocytes, and all intra- and intercellular processes, which may be regulated by miR-29s in the epidermis. I will use human keratinocytes isolated from human skin biopsies remaining after non-therapeutic surgeries (e.g., plastic surgery or circumcisions) and grow keratinocytes into human skin equivalents on artificial collagen matrix. This will allow genetic manipulation of miR-29 levels, biochemical pull-down, purification, and analysis of the functional miR-29 complexes with other molecules. I will study the effect of miR-29 on growth and migration of keratinocytes and will use short synthetic nucleic acid molecules to inhibit miR-29. This will allow development of the new strategy to achieve a successful regeneration of the skin during wound healing, after massive burns, and in reconstructive surgeries. I will also study the function of miR-29s in murine wound healing model, necessary to mimic the complexity of the in vivo regeneration. The long-term goal of the project is to utilize miR-29 to improve skin regeneration in patients suffering from large acute wounds, bedsores, and diabetic ulcers.

在我们的社会中，非愈合性皮肤溃疡的发病率和改善皮肤再生的需求不断增加。为了开发有效的策略来改善皮肤修复，必须了解正常和受损愈合的机制。在我的项目中，我正在寻求进一步描述小核糖核酸（RNA）分子miR-29的功能，我将其确定为皮肤顶层（称为表皮）的调节因子。在受伤时，表皮细胞（主要是角质形成细胞）负责用新的皮肤层覆盖伤口部位，保护我们的身体免受感染和水分流失。角质形成细胞只能在健康皮肤中充分生长并迁移到伤口部位。了解这一过程在正常皮肤中是如何调节的，以及在伤口愈合受损的患者中角质形成细胞的再生是如何失调的，这一点非常重要。本研究的总体目标是促进角质形成细胞的正常生长，从而促进表皮的再生。然而，另一方面，角质形成细胞的过度生长导致皮肤过度增厚、表皮破裂、水分流失和严重的继发性感染。因此，重要的是找到调节和确保对角质形成细胞正常生长的微调控制的分子。小RNA，如miR-29，已经被描述为正常皮肤生长的精确和生理调节剂。重要的是，由于短核酸的小尺寸和化学性质，miR可用于分子治疗。miR-29已经在临床上尝试改善皮肤硬化患者的皮肤状况（表现为异常皮肤增厚），因此可能迅速用于其他皮肤疾病的分子治疗，包括伤口。然而，它需要知道所有可能与角质形成细胞内的miR-29相互作用的分子，以及所有细胞内和细胞间的过程，这些过程可能受到表皮中miR-29的调节。我将使用从非治疗性手术后剩余的人皮肤活检中分离的人角质形成细胞（例如，整形手术或包皮环切术），并在人工胶原基质上将角质形成细胞生长成人类皮肤等同物。这将允许miR-29水平的遗传操作、生化下拉、纯化和功能性miR-29与其他分子的复合物的分析。我将研究miR-29对角质形成细胞生长和迁移的影响，并将使用短的合成核酸分子来抑制miR-29。这将允许开发新的策略，以在伤口愈合期间，大面积烧伤后和重建手术中实现皮肤的成功再生。我还将研究miR-29在小鼠伤口愈合模型中的功能，这是模拟体内再生复杂性所必需的。该项目的长期目标是利用miR-29来改善患有大型急性伤口，褥疮和糖尿病溃疡的患者的皮肤再生。

项目成果

A Nrf2-OSGIN1&2-HSP70 axis mediates cigarette smoke-induced endothelial detachment: implications for plaque erosion.

• DOI: 10.1093/cvr/cvad022
• 发表时间: 2023-08-07
• 期刊: Cardiovascular research
• 影响因子: 10.8

Interaction of the NRF2 and p63 transcription factors promotes keratinocyte proliferation in the epidermis.

• DOI: 10.1093/nar/gkab167
• 发表时间: 2021-04-19
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Kurinna S;Seltmann K;Bachmann AL;Schwendimann A;Thiagarajan L;Hennig P;Beer HD;Mollo MR;Missero C;Werner S
• 通讯作者: Werner S

Spatio-temporal regulation of gene expression defines subpopulations of epidermal stem cells.

• DOI: 10.1042/bst20200740
• 发表时间: 2020-11
• 期刊: Biochemical Society transactions
• 影响因子: 3.9
• 作者: Maneesha Aruketty;S. Kurinna

miR targetome of primary human keratinocytes reveals a function for non-conserved binding sites

原代人角质形成细胞的 miR 靶标揭示了非保守结合位点的功能

• DOI: 10.1101/2022.07.04.498673
• 发表时间: 2022
• 作者: Thiagarajan L