Microvesicles as Potential Biomarkers for the Identification of Senescence in Human Mesenchymal Stem Cells

Abstract

Senescence in human mesenchymal stem cells (MSCs) not only contributes to organism aging and the development of a variety of diseases but also severely impairs their therapeutic properties as a promising cell therapy. Studies searching for efficient biomarkers that represent cellular senescence have attracted much attention; however, no single marker currently provides an accurate cell-free representation of cellular senescence. Here, we studied characteristics of MSC-derived microvesicles (MSC-MVs) that may reflect the senescence in their parental MSCs. We found that senescent late passage (LP) MSCs secreted higher levels of MSC-MVs with smaller size than did early passage (EP) MSCs, and the level of CD105+ MSC-MVs decreased with senescence in the parental MSCs. Also, a substantially weaker ability to promote osteogenesis in MSCs was observed in LP than EP MSC-MVs. Comparative analysis of RNA sequencing showed the same trend of decreasing number of highly-expressed miRNAs with increasing number of passages in both MSCs and MSC-MVs. Most of the highly-expressed genes in LP MSCs and the corresponding MSC-MVs were involved in the regulation of senescence-related diseases, such as Alzheimer's disease. Furthermore, based on the miRNA profiling, transcription factors (TF) and genes regulatory networks of MSC senescence, and the datasets from GEO database, we confirmed that expression of miR-146a-5p in MSC-MVs resembled the senescent state of their parental MSCs. Our findings provide evidence that MSC-MVs are a key factor in the senescence-associated secretory phenotype of MSCs and demonstrate that their integrated characteristics can dynamically reflect the senescence state of MSCs representing a potential biomarker for monitoring MSC senescence.

Highly Purified Human Extracellular Vesicles Produced by Stem Cells Alleviate Aging Cellular Phenotypes of Senescent Human Cells

“…Recently, EVs produced from cultured MSCs were used to rejuvenate aged hematopoietic stem cells or MSCs . It was reported that young (p2–p6) MSCs produced EVs that could rejuvenate aged murine hematopoietic stem cells in a mouse model , similar to a previous report using MSC‐EVs and an irradiation injury mouse model .…”

Involvement of miR‐146a‐5p/neurogenic locus notch homolog protein 1 in the proliferation and differentiation of STRO‐1+ human dental pulp stem cells

“…miR-720 has been reported to promote odontogenic differentiation and reduce proliferation of dental pulp stem/progenitor cells by repressing Nanog homeobox (NANOG) (10). miR-146a-5p (previously known as miR-146a), a member of the miR-146 miRNA family, has been confirmed to play a significant role in the regulation of stem cell proliferation, differentiation, and apoptosis (11)(12)(13). However, this study is the first to demonstrate that miR-146a-5p is involved in the maintenance of the STRO-1 + DPSC state.…”

“…Several miRNAs involved in human‐derived dental stem‐cell maintenance and function have been identified . Recently, it has been reported that miR‐146a‐5p is related to the senescence of human mesenchymal stem cells (hMSCs) and enhances the secretion of vascular endothelial growth factor and the angiogenesis of hMSCs . Overexpression of miR‐146 inhibits the proliferation and promotes the differentiation of mouse neural stem cells .…”

Mesenchymal Stem Cell Derived Extracellular Vesicles in Aging

“…A significant decrease was confirmed for miR-294 and miR-872-3p. In another study, miR-146a and miR-335-5p were up-and down-regulated, respectively, in late passage MSC-EVs versus early passage MSC-EVs but the expression of both miRNAs increased with increasing age in MSC-EVs (Lei et al, 2017). However, the modulated expression of these miRNAs in MSC-EVs was not demonstrated to be related to senescence induction.…”

“…Increase of EV production is a general feature in aged and senescence-induced cells as shown by several studies as early as 2008 (Lehmann et al, 2008;Beer et al, 2015;Takasugi et al, 2017). Accordingly, the production of EVs by MSCs increases with donor age and late passage cultures (Fafian-Labora et al, 2017Lei et al, 2017). The release of EVs from senescent cells is at least partially dependent on p53 and its downstream target gene tumor suppression-activated pathway 6 (TSAP6).…”