Connexin 43 regulates intercellular mitochondrial transfer from human mesenchymal stromal cells to chondrocytes.

IF 7.1 2区 医学 Q1 CELL & TISSUE ENGINEERING Stem Cell Research & Therapy Pub Date : 2024-10-10 DOI:10.1186/s13287-024-03932-9
Rebecca M Irwin, Matthew A Thomas, Megan J Fahey, María D Mayán, James W Smyth, Michelle L Delco
{"title":"Connexin 43 regulates intercellular mitochondrial transfer from human mesenchymal stromal cells to chondrocytes.","authors":"Rebecca M Irwin, Matthew A Thomas, Megan J Fahey, María D Mayán, James W Smyth, Michelle L Delco","doi":"10.1186/s13287-024-03932-9","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>The phenomenon of intercellular mitochondrial transfer from mesenchymal stromal cells (MSCs) has shown promise for improving tissue healing after injury and has potential for treating degenerative diseases like osteoarthritis (OA). Recently MSC to chondrocyte mitochondrial transfer has been documented, but the mechanism of transfer is unknown. Full-length connexin 43 (Cx43, encoded by GJA1) and the truncated, internally translated isoform GJA1-20k have been implicated in mitochondrial transfer between highly oxidative cells, but have not been explored in orthopaedic tissues. Here, our goal was to investigate the role of Cx43 in MSC to chondrocyte mitochondrial transfer. In this study, we tested the hypotheses that (a) mitochondrial transfer from MSCs to chondrocytes is increased when chondrocytes are under oxidative stress and (b) MSC Cx43 expression mediates mitochondrial transfer to chondrocytes.</p><p><strong>Methods: </strong>Oxidative stress was induced in immortalized human chondrocytes using tert-Butyl hydroperoxide (t-BHP) and cells were evaluated for mitochondrial membrane depolarization and reactive oxygen species (ROS) production. Human bone-marrow derived MSCs were transduced for mitochondrial fluorescence using lentiviral vectors. MSC Cx43 expression was knocked down using siRNA or overexpressed (GJA1 + and GJA1-20k+) using lentiviral transduction. Chondrocytes and MSCs were co-cultured for 24 h in direct contact or separated using transwells. Mitochondrial transfer was quantified using flow cytometry. Co-cultures were fixed and stained for actin and Cx43 to visualize cell-cell interactions during transfer.</p><p><strong>Results: </strong>Mitochondrial transfer was significantly higher in t-BHP-stressed chondrocytes. Contact co-cultures had significantly higher mitochondrial transfer compared to transwell co-cultures. Confocal images showed direct cell contacts between MSCs and chondrocytes where Cx43 staining was enriched at the terminal ends of actin cellular extensions containing mitochondria in MSCs. MSC Cx43 expression was associated with the magnitude of mitochondrial transfer to chondrocytes; knocking down Cx43 significantly decreased transfer while Cx43 overexpression significantly increased transfer. Interestingly, GJA1-20k expression was highly correlated with incidence of mitochondrial transfer from MSCs to chondrocytes.</p><p><strong>Conclusions: </strong>Overexpression of GJA1-20k in MSCs increases mitochondrial transfer to chondrocytes, highlighting GJA1-20k as a potential target for promoting mitochondrial transfer from MSCs as a regenerative therapy for cartilage tissue repair in OA.</p>","PeriodicalId":21876,"journal":{"name":"Stem Cell Research & Therapy","volume":"15 1","pages":"359"},"PeriodicalIF":7.1000,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11468299/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Stem Cell Research & Therapy","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1186/s13287-024-03932-9","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CELL & TISSUE ENGINEERING","Score":null,"Total":0}
引用次数: 0

Abstract

Background: The phenomenon of intercellular mitochondrial transfer from mesenchymal stromal cells (MSCs) has shown promise for improving tissue healing after injury and has potential for treating degenerative diseases like osteoarthritis (OA). Recently MSC to chondrocyte mitochondrial transfer has been documented, but the mechanism of transfer is unknown. Full-length connexin 43 (Cx43, encoded by GJA1) and the truncated, internally translated isoform GJA1-20k have been implicated in mitochondrial transfer between highly oxidative cells, but have not been explored in orthopaedic tissues. Here, our goal was to investigate the role of Cx43 in MSC to chondrocyte mitochondrial transfer. In this study, we tested the hypotheses that (a) mitochondrial transfer from MSCs to chondrocytes is increased when chondrocytes are under oxidative stress and (b) MSC Cx43 expression mediates mitochondrial transfer to chondrocytes.

Methods: Oxidative stress was induced in immortalized human chondrocytes using tert-Butyl hydroperoxide (t-BHP) and cells were evaluated for mitochondrial membrane depolarization and reactive oxygen species (ROS) production. Human bone-marrow derived MSCs were transduced for mitochondrial fluorescence using lentiviral vectors. MSC Cx43 expression was knocked down using siRNA or overexpressed (GJA1 + and GJA1-20k+) using lentiviral transduction. Chondrocytes and MSCs were co-cultured for 24 h in direct contact or separated using transwells. Mitochondrial transfer was quantified using flow cytometry. Co-cultures were fixed and stained for actin and Cx43 to visualize cell-cell interactions during transfer.

Results: Mitochondrial transfer was significantly higher in t-BHP-stressed chondrocytes. Contact co-cultures had significantly higher mitochondrial transfer compared to transwell co-cultures. Confocal images showed direct cell contacts between MSCs and chondrocytes where Cx43 staining was enriched at the terminal ends of actin cellular extensions containing mitochondria in MSCs. MSC Cx43 expression was associated with the magnitude of mitochondrial transfer to chondrocytes; knocking down Cx43 significantly decreased transfer while Cx43 overexpression significantly increased transfer. Interestingly, GJA1-20k expression was highly correlated with incidence of mitochondrial transfer from MSCs to chondrocytes.

Conclusions: Overexpression of GJA1-20k in MSCs increases mitochondrial transfer to chondrocytes, highlighting GJA1-20k as a potential target for promoting mitochondrial transfer from MSCs as a regenerative therapy for cartilage tissue repair in OA.

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
连接蛋白43调控从人间质基质细胞到软骨细胞的细胞间线粒体转移。
背景:间充质基质细胞(MSCs)的细胞间线粒体转移现象有望改善损伤后的组织愈合,并具有治疗骨关节炎(OA)等退行性疾病的潜力。最近有记录显示间充质干细胞向软骨细胞转移线粒体,但转移机制尚不清楚。全长的连接蛋白 43(Cx43,由 GJA1 编码)和截短的内部翻译异构体 GJA1-20k 与高度氧化细胞之间的线粒体转移有关,但尚未在骨科组织中进行过研究。在这里,我们的目标是研究 Cx43 在间充质干细胞向软骨细胞线粒体转移中的作用。在这项研究中,我们检验了以下假设:(a)当软骨细胞处于氧化应激时,间充质干细胞向软骨细胞的线粒体转移增加;(b)间充质干细胞 Cx43 的表达介导了向软骨细胞的线粒体转移:方法:使用叔丁基过氧化氢(t-BHP)诱导永生化人软骨细胞产生氧化应激,并评估细胞的线粒体膜去极化和活性氧(ROS)产生情况。使用慢病毒载体转导人骨髓间充质干细胞以获得线粒体荧光。使用 siRNA 敲低间充质干细胞 Cx43 的表达,或使用慢病毒转导过表达(GJA1 + 和 GJA1-20k+)。软骨细胞和间充质干细胞直接接触共培养 24 小时,或使用转孔分离。使用流式细胞术对线粒体转移进行量化。对共培养物进行固定并用肌动蛋白和 Cx43 染色,以观察转移过程中细胞与细胞之间的相互作用:结果:线粒体转移在 t-BHP 受激软骨细胞中明显增加。接触共培养的线粒体转移率明显高于经孔共培养。共聚焦图像显示间充质干细胞和软骨细胞之间有直接的细胞接触,在间充质干细胞中含有线粒体的肌动蛋白细胞延伸末端富集了Cx43染色。间充质干细胞 Cx43 的表达与线粒体向软骨细胞转移的程度有关;敲除 Cx43 会显著减少线粒体向软骨细胞的转移,而过表达 Cx43 则会显著增加线粒体向软骨细胞的转移。有趣的是,GJA1-20k的表达与间充质干细胞向软骨细胞转移线粒体的发生率高度相关:结论:GJA1-20k在间充质干细胞中的过表达可增加线粒体向软骨细胞的转移,这表明GJA1-20k是促进间充质干细胞线粒体转移的潜在靶点,可作为修复OA软骨组织的再生疗法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
Stem Cell Research & Therapy
Stem Cell Research & Therapy CELL BIOLOGY-MEDICINE, RESEARCH & EXPERIMENTAL
CiteScore
13.20
自引率
8.00%
发文量
525
审稿时长
1 months
期刊介绍: Stem Cell Research & Therapy serves as a leading platform for translational research in stem cell therapies. This international, peer-reviewed journal publishes high-quality open-access research articles, with a focus on basic, translational, and clinical research in stem cell therapeutics and regenerative therapies. Coverage includes animal models and clinical trials. Additionally, the journal offers reviews, viewpoints, commentaries, and reports.
期刊最新文献
Exosome crosstalk between cancer stem cells and tumor microenvironment: cancer progression and therapeutic strategies. Reprogrammed human lateral ganglionic eminence precursors generate striatal neurons and restore motor function in a rat model of Huntington's disease. Rapid-acting pain relief in knee osteoarthritis: autologous-cultured adipose-derived mesenchymal stem cells outperform stromal vascular fraction: a systematic review and meta-analysis. Comparative analysis of regulations and studies on stem cell therapies: focusing on induced pluripotent stem cell (iPSC)-based treatments. Correction: Adipose stem cells regulate lipid metabolism by upregulating mitochondrial fatty acid β-oxidation in macrophages to improve the retention rate of transplanted fat.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1