MEG3 shuttled by exosomes released from human bone marrow mesenchymal stem cells promotes TP53 stability to regulate MCM5 transcription in keloid fibroblasts

IF 3.2 4区医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of Gene Medicine Pub Date : 2024-04-30 DOI:10.1002/jgm.3688

Feibin Zhu, Yuanjian Ye, Ying Shao, Chunli Xue

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Abstract

Background

Despite the interest in mesenchymal stem cells (MSC), their potential to treat abnormal scarring, especially keloids, is yet to be described. The present study aimed to investigate the therapeutic potential of exosomes derived from human bone marrow MSCs (hBMSC-Exos) in alleviating keloid formation.

Methods

Exosomes were isolated from hBMSC, and keloid fibroblasts (KFs) were treated with hBMSC-Exos. Cell counting kit-8, wound healing, transwell invasion, immunofluorescence, and western blot assays were conducted to study the malignant phenotype of KFs. Mice were induced with keloids and treated with hBMSC-Exos. The effect of hBMSC-Exos on keloid formation in vivo was evaluated by hematoxylin and eosin staining, Masson staining, immunohistochemistry, and western blotting. The GSE182192 dataset was screened for differentially expressed long non-coding RNA during keloid formation. Next, maternally expressed gene 3 (MEG3) was knocked down in hBMSC to obtain hBMSC-Exos^sh-MEG3. The molecular mechanism of MEG3 was investigated by bioinformatic screening, and the relationship between MEG3 and TP53 or MCM5 was verified.

Results

hBMSC-Exos inhibited the malignant proliferation, migration, and invasion of KFs at same time as promoting their apoptosis, Moreover, hBMSC-Exos reduced the expression of fibrosis- and collagen-related proteins in the cells and the formation of keloids caused by KFs. The reduction in MEG3 enrichment in hBMSC-Exos weakened the inhibitory effect of hBMSC-Exos on KF activity. hBMSC-Exos delivered MEG3 to promote MCM5 transcription by TP53 in KFs. Overexpression of MCM5 in KFs reversed the effects of hBMSC-Exos^sh-MEG3, leading to reduced KF activity.

Conclusions

hBMSC-Exos delivered MEG3 to promote the protein stability of TP53, thereby activating MCM5 and promoting KF activity.

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人骨髓间充质干细胞释放的外泌体穿梭的 MEG3 可促进 TP53 的稳定性，从而调节瘢痕疙瘩成纤维细胞中 MCM5 的转录

背景尽管间充质干细胞（MSC）备受关注，但其治疗异常瘢痕，尤其是瘢痕疙瘩的潜力尚未得到描述。本研究旨在探讨从人类骨髓间充质干细胞中提取的外泌体（hBMSC-Exos）在缓解瘢痕疙瘩形成方面的治疗潜力。方法从 hBMSC 中分离出外泌体，用 hBMSC-Exos 处理瘢痕疙瘩成纤维细胞（KFs）。通过细胞计数试剂盒-8、伤口愈合、Transwell侵袭、免疫荧光和Western印迹检测来研究KFs的恶性表型。诱导小鼠患瘢痕疙瘩，并用 hBMSC-Exos 治疗。通过苏木精和伊红染色、Masson 染色、免疫组织化学和 Western 印迹法评估了 hBMSC-Exos 对体内瘢痕疙瘩形成的影响。在 GSE182192 数据集中筛选了瘢痕疙瘩形成过程中差异表达的长非编码 RNA。然后，在 hBMSC 中敲除母源表达基因 3（MEG3），得到 hBMSC-Exossh-MEG3。通过生物信息学筛选研究了 MEG3 的分子机制，并验证了 MEG3 与 TP53 或 MCM5 的关系。结果 hBMSC-Exos 可抑制 KFs 的恶性增殖、迁移和侵袭，同时促进其凋亡。hBMSC-Exos 中 MEG3 富集的减少削弱了 hBMSC-Exos 对 KF 活性的抑制作用。在 KFs 中过表达 MCM5 可逆转 hBMSC-Exossh-MEG3 的作用，从而降低 KF 的活性。结论 hBMSC-Exos 传递 MEG3 促进 TP53 蛋白的稳定性，从而激活 MCM5 并促进 KF 的活性。

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来源期刊

Journal of Gene Medicine 医学-生物工程与应用微生物

CiteScore

6.40

自引率

0.00%

发文量

审稿时长

6-12 weeks

期刊介绍： The aims and scope of The Journal of Gene Medicine include cutting-edge science of gene transfer and its applications in gene and cell therapy, genome editing with precision nucleases, epigenetic modifications of host genome by small molecules, siRNA, microRNA and other noncoding RNAs as therapeutic gene-modulating agents or targets, biomarkers for precision medicine, and gene-based prognostic/diagnostic studies. Key areas of interest are the design of novel synthetic and viral vectors, novel therapeutic nucleic acids such as mRNA, modified microRNAs and siRNAs, antagomirs, aptamers, antisense and exon-skipping agents, refined genome editing tools using nucleic acid /protein combinations, physically or biologically targeted delivery and gene modulation, ex vivo or in vivo pharmacological studies including animal models, and human clinical trials. Papers presenting research into the mechanisms underlying transfer and action of gene medicines, the application of the new technologies for stem cell modification or nucleic acid based vaccines, the identification of new genetic or epigenetic variations as biomarkers to direct precision medicine, and the preclinical/clinical development of gene/expression signatures indicative of diagnosis or predictive of prognosis are also encouraged.