Ma Hongrui, Cao Yashi, Gao Zizheng, Xu Zhifei, Yang Bo, Luo Peihua, Hu Yuhuai, He Qiaojun
Center for Drug Safety Evaluation and Research of Zhejiang University, College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Zijingang Campus, Hangzhou, 310058, Zhejiang, PR China.
Institute of Pharmacology & Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, PR China.
Stem Cell Rev Rep. 2025 Jun 28. doi: 10.1007/s12015-025-10929-1.
The limited regenerative capacity of epidermal cells following tissue injury impairs wound healing in diabetic foot ulcers (DFUs), contributing to elevated rates of amputation and mortality. Recent advances have demonstrated that somatic cells can be reprogrammed into diverse cell types through the application of defined reprogramming factors. This study aims to develop a safe, efficient, and clinically translatable strategy for skin regeneration via direct lineage reprogramming.
We established a novel reprogramming approach using a combination of two factors, BMI1 and FGFR2b (termed B2), to induce fibroblast-to-keratinocyte-like cells (iKCs) conversion in vitro and delivered via adeno-associated virus 9 (AAV9) in vivo. Molecular and functional characteristics of iKCs were evaluated by qRT-PCR, Western blot, immunofluorescence, transcriptomic analysis, and in vitro differentiation assays. A diabetic (db/db) mouse skin wound model was used to assess the regenerative potential and therapeutic effects. Statistical significance was determined using Student's t-test or one-way ANOVA.
iKCs-B2 (Keratinocyte-like cells form from B2-infected L929) exhibited both morphological and functional characteristics comparable to primary keratinocytes. In vivo, AAV9-mediated delivery of B2 factors significantly promoted wound closure, reconstructed stratified epithelium, restored barrier function, and markedly reduced the mortality rate.
This study presents a safe and effective direct reprogramming strategy for skin regeneration, bypassing the pluripotent stage and avoiding cell transplantation. The B2 combination provides a novel molecular tool for wound repair and offers translational potential for treating non-healing wounds such as DFUs.
组织损伤后表皮细胞有限的再生能力会损害糖尿病足溃疡（DFU）的伤口愈合，导致截肢率和死亡率升高。最近的进展表明，通过应用特定的重编程因子，体细胞可以被重编程为多种细胞类型。本研究旨在通过直接谱系重编程开发一种安全、高效且可临床转化的皮肤再生策略。
我们建立了一种新的重编程方法，使用BMI1和FGFR2b这两种因子的组合（称为B2），在体外诱导成纤维细胞向角质形成细胞样细胞（iKC）转化，并通过腺相关病毒9（AAV9）在体内递送。通过qRT-PCR、蛋白质免疫印迹、免疫荧光、转录组分析和体外分化试验评估iKC的分子和功能特征。使用糖尿病（db/db）小鼠皮肤伤口模型评估再生潜力和治疗效果。使用学生t检验或单因素方差分析确定统计学意义。
iKCs-B2（由B2感染的L929形成的角质形成细胞样细胞）表现出与原代角质形成细胞相当的形态和功能特征。在体内，AAV9介导的B2因子递送显著促进伤口闭合、重建分层上皮、恢复屏障功能，并显著降低死亡率。
本研究提出了一种安全有效的皮肤再生直接重编程策略，绕过了多能阶段，避免了细胞移植。B2组合为伤口修复提供了一种新的分子工具，并为治疗DFU等不愈合伤口提供了转化潜力。
