m6A阅读蛋白YTHDF2调控ACSL4介导糖尿病角质形成细胞铁死亡及其在伤口愈合中的作用机制研究

糖尿病皮肤角质形成细胞减少，皮肤变薄，伤口难愈，机制有待研究。我们发现糖尿病角质形成细胞存在铁死亡：其铁死亡关键蛋白ACSL4、及脂过氧化增加，同时伴有m6A阅读蛋白YTHDF2减少。YTHDF2功能是促进m6A修饰的mRNA降解，那么糖尿病状态下YTHDF2是否能调控ACSL4的表达，导致角质形成细胞铁死亡，目前尚不清楚？且我们已证明YTHDF2能结合m6A修饰的ACSL4 mRNA。因此我们假设糖尿病状态下，角质形成细胞下调的YTHDF2，使ACSL4表达增加，引起细胞发生铁死亡，导致皮肤易损难愈，而过表达YTHDF2或铁死亡抑制剂均可以挽救细胞铁死亡促进伤口愈合。故本项目拟采用慢病毒过表达及敲低YTHDF2、RIP-qPCR等方法来阐明YTHDF2调控m6A修饰的ACSL4 介导糖尿病角质形成细胞铁死亡及其在糖尿病伤口愈合中的作用，为糖尿病皮肤伤口治疗的提供新思路。

The diabetic keratinocytes were fewer and thickness of skin was thinner，which leads to poor wound healing in diabetes. The underlying mechanisms remain largely unknown. We provided evidence showing both the acyl-CoA synthetase long-chain family member 4（ACSL4）（a key regulator of ferroptosis）and lipid peroxidation products were increased, while m6A reader YTHDF2 was reduced in diabetic keratinocytes. And YTHDF2 was reported to regulate degradation of m6A-modifed mRNAs. However, whether YTHDF2 can mediate the stability of ACSL4 mRNA is still unknown? We found that ACSL4 mRNA was modified by m6A and bound by YTHDF2. So we hypothesize that decreased YTHDF2 can increase the expression of ACSL4, thus causing ferroptosis of keratinocytes and impairing wound healing during diabetic condition. Specific overexpression of YTHDF2 and application of ferroptosis inhibitors could promote wound healing through inhibiting ferroptosis of keratinocytes. Therefore, Molecular and cellular techniques, including lentiviral vector-mediated knockdown or overexpression and RIP-qPCR will be used to elucidate the mechanism that YTHDF2-regulated m6A modification of ACSL4 mediates ferroptosis in diabetic keratinocytes, so as to provide novel therapeutic targets and strategies for improving diabetic wound healing.