Human mesenchymal stromal cells release functional mitochondria in extracellular vesicles.
Human mesenchymal stromal cells release functional mitochondria in extracellular vesicles.
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人类间充质基质细胞在细胞外囊泡中释放功能性线粒体。
DOI：
10.3389/fbioe.2022.870193
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发表时间：
2022
影响因子：
5.7
通讯作者：
Delco, Michelle L.
中科院分区：
文献类型：
--
作者：
Thomas, Matthew A.;Fahey, Megan J.;Pugliese, Brenna R.;Irwin, Rebecca M.;Antonyak, Marc A.;Delco, Michelle L.
软骨和其他骨骼软组织在损伤后愈合不良，部分原因是它们缺乏血管分布和低代谢率。没有药理学方法已被证明可以有效预防关节损伤后的慢性退行性疾病。间充质基质细胞（MSC）已被研究其治疗与骨关节炎（OA）相关的疼痛和保护关节软骨的能力。MSC的局限性包括细胞表型的可变性、低植入率和保留率以及不一致的临床结果。因此，目前正在研究诸如细胞外囊泡（EV）的无细胞生物疗法。已经发现MSC衍生的EV复制了其来源细胞的许多治疗效果，但驱动这一点的机制仍不清楚。最近在非骨科组织中的证据表明，MSC可以通过捐献线粒体、恢复受体细胞中的线粒体功能、保持细胞活力和促进组织修复来拯救受损细胞。我们的研究小组假设MSC包装线粒体以出口到EV中，并且这些所谓的“mitoEV”可以为无细胞的靶向治疗提供递送策略。因此，本研究的目的是：1）表征MSC分泌组的囊泡部分相对于线粒体货物，2）确定MSC-EV是否含有功能性线粒体，和3）确定软骨细胞是否可以摄取MSC衍生的mitoEV。我们从MSC条件培养基中分离出外泌体、微囊泡和无囊泡组分。使用动态光散射和纳米颗粒跟踪的组合，我们确定MSC-EV群体属于通常用于对EV进行分类的三个大小类别（外来体、微泡、凋亡小体）。荧光纳米颗粒追踪、免疫印迹和流式细胞术显示，线粒体货物在所有EV大小的群体中都是丰富的，并且mitoEV在最大的EV中几乎无处不在。极化染色表明mitoEV的一个子集含有功能性线粒体。最后，流式细胞术和荧光成像证实了线粒体EV的软骨细胞进行鱼藤酮/抗霉素诱导的线粒体功能障碍的摄取。这些数据表明MSC将完整的功能性线粒体包装成EV，其可以在没有直接细胞-细胞相互作用的情况下转移到软骨细胞。这项工作表明，健康MT向软骨细胞的细胞间转移可能代表一种新的非细胞方法，以增加愈合不良的无血管骨骼软组织中的线粒体含量和功能。
Cartilage and other skeletal soft tissues heal poorly after injury, in part due to their lack of vascularity and low metabolic rate. No pharmacologic approaches have proven effective in preventing chronic degenerative disease after joint injury. Mesenchymal stromal cells (MSCs) have been investigated for their ability to treat pain associated with osteoarthritis (OA) and preserve articular cartilage. Limitations of MSCs include variability in cell phenotype, low engraftment and retention rates, and inconsistent clinical outcomes. Therefore, acellular biologic therapies such as extracellular vesicles (EVs) are currently being investigated. MSC-derived EVs have been found to replicate many of the therapeutic effects of their cells of origin, but the mechanisms driving this remain unclear. Recent evidence in non-orthopedic tissues suggests MSCs can rescue injured cells by donating mitochondria, restoring mitochondrial function in recipient cells, preserving cell viability, and promoting tissue repair. Our group hypothesized that MSCs package mitochondria for export into EVs, and that these so-called “mitoEVs” could provide a delivery strategy for cell-free mitochondria-targeted therapy. Therefore, the goals of this study were to: 1) characterize the vesicle fractions of the MSCs secretome with respect to mitochondrial cargoes, 2) determine if MSC-EVs contain functional mitochondria, and 3) determine if chondrocytes can take up MSC-derived mitoEVs. We isolated exosome, microvesicle, and vesicle-free fractions from MSC-conditioned media. Using a combination of dynamic light scattering and nanoparticle tracking, we determined that MSC-EV populations fall within the three size categories typically used to classify EVs (exosomes, microvesicles, apoptotic bodies). Fluorescent nanoparticle tracking, immunoblotting, and flow cytometry revealed that mitochondrial cargoes are abundant across all EV size populations, and mitoEVs are nearly ubiquitous among the largest EVs. Polarization staining indicated a subset of mitoEVs contain functional mitochondria. Finally, flow cytometry and fluorescent imaging confirmed uptake of mitoEVs by chondrocytes undergoing rotenone/antimycin-induced mitochondrial dysfunction. These data indicate that MSCs package intact, functional mitochondria into EVs, which can be transferred to chondrocytes in the absence of direct cell-cell interactions. This work suggests intercellular transfer of healthy MT to chondrocytes could represent a new, acellular approach to augment mitochondrial content and function in poorly-healing avascular skeletal soft tissues.
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影响因子：
29
作者：
Crewe C;Funcke JB;Li S;Joffin N;Gliniak CM;Ghaben AL;An YA;Sadek HA;Gordillo R;Akgul Y;Chen S;Samovski D;Fischer-Posovszky P;Kusminski CM;Klein S;Scherer PE
通讯作者：
Scherer PE
影响因子：
3.3
作者：
Sophia Fox AJ;Bedi A;Rodeo SA
通讯作者：
Rodeo SA
影响因子：
2.8
作者：
Bartell, Lena R.;Fortier, Lisa A.;Delco, Michelle L.
通讯作者：
Delco, Michelle L.
影响因子：
2.8
作者：
Goetz, Jessica E.;Coleman, Mitchell C.;Fredericks, Douglas C.;Petersen, Emily;Martin, James A.;McKinley, Todd O.;Tochigi, Yuki
通讯作者：
Tochigi, Yuki
影响因子：
4.7
作者：
Asghar, Waseem;El Assal, Rami;Shafiee, Hadi;Anchan, Raymond M.;Demirci, Utkan
通讯作者：
Demirci, Utkan