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

Yan Shi; Shang Wang; Dewu Liu; Zhengguang Wang; Yihan Zhu; Jun Li; Kui Xu; Furong Li; Huicai Wen; Ronghua Yang

doi:10.1093/burnst/tkad058

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

更新时间：2024-04-19

- Exosomal miR-4645-5p from hypoxic bone marrow mesenchymal stem cells facilitates diabetic wound healing by restoring keratinocyte autophagy
- Exosomal miR-4645-5p from hypoxic bone marrow mesenchymal stem cells facilitates diabetic wound healing by restoring keratinocyte autophagy
- Burns & Trauma 2024年12卷第1期
- 作者机构：
  
  1. Department of Plastic
  2. The First Affiliated Hospital, Jiangxi Medical College, Nanchang University
  3. Chongqing Key Laboratory of Traditional Chinese Medicine for Prevention and Cure of Metabolic Diseases, College of Traditional Chinese Medicine, Chongqing Medical University
  4. Medical Center of Burn Plastic and Wound Repair, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University
  5. Department of Orthopaedics, Peking University Third Hospital
  6. Department of Plastic and Aesthetic Surgery, Jiangxi Maternal and Child Health Hospital
  7. HaploX Biotechnology Co
  8. Key Laboratory of Xin’an Medicine
  9. Anhui University of Chinese Medicine
  10. Translational Medicine Collaborative Innovation Center, Shenzhen People’s Hospital (The Second Clinical Medical College, Jinan University
  11. The First Affifiliated Hospital, Southern University of Science and Technology
  12. Department of Burn and Plastic Surgery
  13. South China University of Technology
- 作者简介：
- 基金信息：
  
  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), GuangDong 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).
- DOI：10.1093/burnst/tkad058
  中图分类号：
- 纸质出版日期：2024，
  
  网络出版日期：2024-1-17，
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Yan Shi, Shang Wang, Dewu Liu, 等. Exosomal miR-4645-5p from hypoxic bone marrow mesenchymal stem cells facilitates diabetic wound healing by restoring keratinocyte autophagy[J]. Burns & Trauma, 2024,12(1).

Yan Shi, Shang Wang, Dewu Liu, Zhengguang Wang, Yihan Zhu, Jun Li, Kui Xu, Furong Li, Huicai Wen, Ronghua Yang, Exosomal miR-4645-5p from hypoxic bone marrow mesenchymal stem cells facilitates diabetic wound healing by restoring keratinocyte autophagy, Burns & Trauma, Volume 12, 2024, tkad058, https://doi.org/10.1093/burnst/tkad058
Yan Shi, Shang Wang, Dewu Liu, 等. Exosomal miR-4645-5p from hypoxic bone marrow mesenchymal stem cells facilitates diabetic wound healing by restoring keratinocyte autophagy[J]. Burns & Trauma, 2024,12(1). DOI： 10.1093/burnst/tkad058.

Yan Shi, Shang Wang, Dewu Liu, Zhengguang Wang, Yihan Zhu, Jun Li, Kui Xu, Furong Li, Huicai Wen, Ronghua Yang, Exosomal miR-4645-5p from hypoxic bone marrow mesenchymal stem cells facilitates diabetic wound healing by restoring keratinocyte autophagy, Burns & Trauma, Volume 12, 2024, tkad058, https://doi.org/10.1093/burnst/tkad058 DOI：

摘要

Abstract

BackgroundRefractory 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.MethodsWe 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.ResultsHypoxia 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.ConclusionsOverall

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.

关键词

Keywords

Bone marrow mesenchymal stem cellAKT-mTORC1 signalingDiabetic wound repairEpidermal autophagyExosomesMAPKAPK2MigrationProliferationRe-epithelizationkeratinocyte

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