Wound Healing and Regenerative Medicine Research Program, Department of Dermatology & Cutaneous Surgery, University of Miami Miller School of Medicine, Miami, FL 33136, USA.
J Biol Chem. 2012 Aug 24;287(35):29324-35. doi: 10.1074/jbc.M112.382135. Epub 2012 Jul 6.
Chronic nonhealing wounds, such as venous ulcers (VUs), are a widespread and serious medical problem with high morbidity and mortality. The molecular pathology of VUs remains poorly understood, impeding the development of effective treatment strategies. Using mRNA expression profiling of VUs biopsies and computational analysis, we identified a candidate set of microRNAs with lowered target gene expression. Among these candidates, miR-16, -20a, -21, -106a -130a, and -203 were confirmed to be aberrantly overexpressed in a cohort study of 10 VU patients by quantitative PCR and in situ hybridizations. These microRNAs were predicted to target multiple genes important for wound healing, including early growth response factor 3, vinculin, and leptin receptor (LepR). Overexpression of the top up-regulated miRNAs, miR-21 and miR-130a, in primary human keratinocytes down-regulated expression of the endogenous LepR and early growth response factor 3. The luciferase reporter assay verified LepR as a direct target for miR-21 and miR-130a. Both miR-21 and miR-130a delayed epithelialization in an acute human skin wound model. Furthermore, in vivo overexpression of miR-21 inhibited epithelialization and granulation tissue formation in a rat wound model. Our results identify a novel mechanism in which overexpression of specific set of microRNAs inhibits wound healing, resulting in new potential molecular markers and targets for therapeutic intervention.
慢性难愈性创面,如静脉性溃疡(venous ulcers,VUs),是一种广泛存在且严重的医学问题,具有较高的发病率和死亡率。VUs 的分子病理学仍知之甚少,这阻碍了有效治疗策略的发展。通过 VUs 活检的 mRNA 表达谱分析和计算分析,我们确定了一组候选 microRNAs,其靶基因表达降低。在 10 名 VU 患者的队列研究中,通过定量 PCR 和原位杂交,证实了这些候选 microRNAs 中的 miR-16、-20a、-21、-106a-130a 和 -203 异常过表达。这些 microRNAs 被预测靶向多个与伤口愈合相关的重要基因,包括早期生长反应因子 3、纽蛋白和瘦素受体(leptin receptor,LepR)。在原代人角质形成细胞中过表达 top 上调的 microRNAs,miR-21 和 miR-130a,下调了内源性 LepR 和早期生长反应因子 3 的表达。荧光素酶报告基因测定验证了 LepR 是 miR-21 和 miR-130a 的直接靶标。miR-21 和 miR-130a 均延迟了急性人皮肤创面模型中的上皮化。此外,在大鼠创面模型中体内过表达 miR-21 抑制了上皮化和肉芽组织形成。我们的研究结果确定了一种新的机制,即特定 microRNAs 集合的过表达抑制伤口愈合,从而为新的潜在分子标志物和治疗干预靶点提供了依据。