MicroRNA-183-5p Increases with Age in Bone-Derived Extracellular Vesicles, Suppresses Bone Marrow Stromal (Stem) Cell Proliferation, and Induces Stem Cell Senescence

Abstract

Microvesicle- and exosome-mediated transport of microRNAs (miRNAs) represents a novel cellular and molecular pathway for cell-cell communication. In this study, we tested the hypothesis that these extracellular vesicles (EVs) and their miRNAs might change with age, contributing to age-related stem cell dysfunction. EVs were isolated from the bone marrow interstitial fluid (supernatant) of young (3-4 months) and aged (24-28 months) mice to determine whether the size, concentration, and miRNA profile of EVs were altered with age in vivo. Results show that EVs isolated from bone marrow are CD63 and CD9 positive, and the concentration and size distribution of bone marrow EVs are similar between the young and aged mice. Bioanalyzer data indicate that EVs from both young and aged mice are highly enriched in miRNAs, and the miRNA profile of bone marrow EVs differs significantly between the young and aged mice. Specifically, the miR-183 cluster (miR-96/-182/-183) is highly expressed in aged EVs. In vitro assays demonstrate that aged EVs are endocytosed by primary bone marrow stromal cells (BMSCs), and these aged EVs inhibit the osteogenic differentiation of young BMSCs. Transfection of BMSCs with miR-183-5p mimic reduces cell proliferation and osteogenic differentiation, increases senescence, and decreases protein levels of the miR-183-5p target heme oxygenase-1 (Hmox1). In vitro assays utilizing HO-induced oxidative stress show that HO treatment of BMSCs increases the abundance of miR-183-5p in BMSC-derived EVs, and Amplex Red assays demonstrate that HO is elevated in the bone marrow microenvironment with age. Together, these data indicate that aging and oxidative stress can significantly alter the miRNA cargo of EVs in the bone marrow microenvironment, which may in turn play a role in stem cell senescence and osteogenic differentiation by reducing Hmox1 activity.

Senescent cells: Living or dying is a matter of NK cells

“…In general, EVs carry and transfer a wide variety of molecules, such as micro RNAs (miRNAs), messenger RNAs, DNA, and proteins. Remarkably, EVs secreted from senescent cells have unique characteristics and contribute to regulate the behavior of recipient cells similarly to soluble SASP factors . By analyzing EVs from senescent human dermal fibroblasts, Terlecki‐Zaniewicz and colleagues have identified a set of selectively retained or secreted miRNAs and have shown that senescent cell‐derived EVs with their miRNA cargo contribute to an antiapoptotic environment in tissues where senescent cells accumulate .…”

“…Remarkably, EVs secreted from senescent cells have unique characteristics and contribute to regulate the behavior of recipient cells similarly to soluble SASP factors. 56,57 By analyzing EVs from senescent human dermal fibroblasts, Terlecki-Zaniewicz and colleagues have identified a set of selectively retained or secreted miRNAs and have shown that senescent cell-derived EVs with their miRNA cargo contribute to an antiapoptotic environment in tissues where senescent cells accumulate. 58 Furthermore, ovarian cancer-derived exosomes expressing miRNA-433 have the potential to modulate the tumor microenvironment by inducing cellular senescence in neighboring cells.…”

Peripheral blood exosomes pass blood-brain-barrier and induce glial cell activation

“…Plasma EVs isolated from young but not from elderly donors promote the osteogenic differentiation of mesenchymal stem cells in a galectin-3 dependent manner 18 . Furthermore, EVs purified from the elderly suppress cell proliferation and osteogenic differentiation of bone marrow stromal cells 3 .…”

SILAC-based quantitative proteomics reveals pleiotropic, phenotypic modulation in primary murine macrophages infected with the protozoan pathogenLeishmania donovani

“…In our infection system, despite the repressed protein levels, we find Hmox-1 mRNA to be up-regulated in macrophages infected with L. donovani . The negative correlation between Hmox-1 RNA and protein levels, amongst other factors, could be due to increased secretion of Hmox-1 in the culture medium, as Hmox-1 can be a secreted protein [86].…”