Exosomal miR-4645-5p from hypoxic bone marrow mesenchymal stem cells facilitates diabetic wound healing by restoring keratinocyte autophagy

作者全名："Shi, Yan; Wang, Shang; Liu, Dewu; Wang, Zhengguang; Zhu, Yihan; Li, Jun; Xu, Kui; Li, Furong; Wen, Huicai; Yang, Ronghua"

作者地址："[Shi, Yan; Wen, Huicai] Nanchang Univ, Affiliated Hosp 1, Jiangxi Med Coll, Dept Plast, Yongwaizheng Rd, Donghu Dist, Nanchang 330006, Jiangxi, Peoples R China; [Wang, Shang] Chongqing Med Univ, Coll Tradit Chinese Med, Chongqing Key Lab Tradit Chinese Med Prevent & Cur, Med Coll Rd, Chongqing 400016, Peoples R China; [Liu, Dewu] Nanchang Univ, Affiliated Hosp 1, Jiangxi Med Coll, Med Ctr Burn Plast & Wound Repair, Yongwaizheng Rd, Nanchang 330006, Jiangxi, Peoples R China; [Wang, Zhengguang] Peking Univ Third Hosp, Dept Orthopaed, 49 North Garden Rd, Beijing 100191, Peoples R China; [Zhu, Yihan] Jiangxi Maternal & Child Hlth Hosp, Dept Plast & Aesthet Surg, Bayidadao Rd, Nanchang 330006, Peoples R China; [Li, Jun] HaploX Biotechnol Co Ltd, Songpingshan Rd, Shenzhen 518057, Guangdong, Peoples R China; [Xu, Kui] Anhui Univ Chinese Med, Key Lab Xinan Med, Minist Educ, Qianjiang Rd, Hefei 230038, Anhui, Peoples R China; [Li, Furong] Jinan Univ, Shenzhen Peoples Hosp, Clin Med Coll 2, Translat Med Collaborat Innovat Ctr, Guangzhou, Peoples R China; [Li, Furong] Southern Univ Sci & Technol, Affifiliated Hosp 1, Dongmenbei Rd, Shenzhen 518020, Guangdong, Peoples R China; [Yang, Ronghua] South China Univ Technol, Guangzhou Peoples Hosp 1, Dept Burn & Plast Surg, Panfu Rd, Guangzhou 510180, Guangdong, Peoples R China"

通信作者："Wen, HC (通讯作者)，Nanchang Univ, Affiliated Hosp 1, Jiangxi Med Coll, Dept Plast, Yongwaizheng Rd, Donghu Dist, Nanchang 330006, Jiangxi, Peoples R China.; Li, FR (通讯作者)，Jinan Univ, Shenzhen Peoples Hosp, Clin Med Coll 2, Translat Med Collaborat Innovat Ctr, Guangzhou, Peoples R China.; Li, FR (通讯作者)，Southern Univ Sci & Technol, Affifiliated Hosp 1, Dongmenbei Rd, Shenzhen 518020, Guangdong, Peoples R China.; Yang, RH (通讯作者)，South China Univ Technol, Guangzhou Peoples Hosp 1, Dept Burn & Plast Surg, Panfu Rd, Guangzhou 510180, Guangdong, Peoples R China."

来源：BURNS & TRAUMA

ESI学科分类：CLINICAL MEDICINE

WOS号：WOS:001145419400001

JCR分区：Q1

影响因子：5.3

年份：2024

卷号：12

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：Bone marrow mesenchymal stem cell; AKT-mTORC1 signaling; Diabetic wound repair; Epidermal autophagy; Exosomes; MAPKAPK2; Migration; Proliferation; Re-epithelization; Keratinocyte

摘要："Background Refractory diabetic wounds are a common occurrence in patients with diabetes and epidermis-specific macroautophagy/autophagy impairment has been implicated in their pathogenesis. Therefore, identifying and developing treatment strategies capable of normalizing epidermis-specific macroautophagy/autophagy could facilitate diabetic wound healing. The study aims to investigate the potential of bone marrow mesenchymal stem cell-derived exosomes (BMSC-exos) from hypoxic conditions as a treatment to normalize epidermis-specific autophagy for diabetic wound healing.Methods We compared the effects of bone marrow mesenchymal stem cell (BMSC)-sourced exosomes (BMSC-Exos) from hypoxic conditions to those of BMSC in normoxic conditions (noBMSC-Exos). Our studies involved morphometric assessment of the exosomes, identification of the microRNA (miRNA) responsible for the effects, evaluation of keratinocyte functions and examination of effects of the exosomes on several molecules involved in the autophagy pathway such as microtubule-associated protein 1 light chain 3 beta, beclin 1, sequestosome 1, autophagy-related 5 and autophagy-related 5. The experiments used human BMSCs from the American Type Culture Collection, an in vivo mouse model of diabetes (db/db) to assess wound healing, as well as the human keratinocyte HaCaT cell line. In the methodology, the authors utilized an array of approaches that included electron microscopy, small interfering RNA (siRNA) studies, RNA in situ hybridization, quantitative real-time reverse transcription PCR (qRT-PCR), the isolation, sequencing and differential expression of miRNAs, as well as the use of miR-4645-5p-specific knockdown with an inhibitor.Results Hypoxia affected the release of exosomes from hypoxic BMSCs (hy-BMSCs) and influenced the size and morphology of the exosomes. Moreover, hyBMSC-Exo treatment markedly improved keratinocyte function, including keratinocyte autophagy, proliferation and migration. miRNA microarray and bioinformatics analysis showed that the target genes of the differentially expressed miRNAs were mainly enriched in 'autophagy' and 'process utilizing autophagic mechanism' in the 'biological process' category and miR-4645-5p as a major contributor to the pro-autophagy effect of hyBMSC-Exos. Moreover, mitogen-activated protein kinase-activated protein kinase 2 (MAPKAPK2) was identified as a potential target of exosomal miR-4645-5p; this was confirmed using a dual luciferase assay. Exosomal miR-4645-5p mediates the inactivation of the MAPKAPK2-induced AKT kinase group (comprising AKT1, AKT2, and AKT3), which in turn suppresses AKT-mTORC1 signaling, thereby facilitating miR-4645-5p-mediated autophagy.Conclusions Overall, the results of this study showed that hyBMSC-Exo-mediated transfer of miR-4645-5p inactivated MAPKAPK2-induced AKT-mTORC1 signaling in keratinocytes, which activated keratinocyte autophagy, proliferation and migration, resulting in diabetic wound healing in mice. Collectively, the findings could aid in the development of a novel therapeutic strategy for diabetic wounds."

基金机构："National Natural Science Foundation of China [82060350, 82002272, 82272276]; China Postdoctoral Science Foundation [2022 M711335, 2021 M701434]; Guang-Dong Basic and Applied Basic Research Foundation [2022A1515110490, 2022A1515011380, 2022A1515012160]; Industry-university-research Innovation Fund of Higher Education of China [2021JH028]; Science and Technology Innovation Committee of Shenzhen [JCYJ20220530152015036]"

基金资助正文："This study was supported by the National Natural Science Foundation of China (No. 82060350, No. 82002272,No. 82272276), China Postdoctoral Science Foundation (No. 2022 M711335, No. 2021 M701434), Guang-Dong Basic and Applied Basic Research Foundation (No.2022A1515110490, No.2021A1515011453, No.2022A1515011380, No. 2022A1515012160), Industry-university-research Innovation Fund of Higher Education of China (No. 2021JH028), the Science and Technology Innovation Committee of Shenzhen (No. JCYJ20220530152015036)"