TY - JOUR
T1 - MicroRNA-183-5p Increases with Age in Bone-Derived Extracellular Vesicles, Suppresses Bone Marrow Stromal (Stem) Cell Proliferation, and Induces Stem Cell Senescence
AU - Davis, Colleen
AU - Dukes, Amy
AU - Drewry, Michelle
AU - Helwa, Inas
AU - Johnson, Maribeth H.
AU - Isales, Carlos M.
AU - Hill, William D.
AU - Liu, Yutao
AU - Shi, Xingming
AU - Fulzele, Sadanand
AU - Hamrick, Mark W.
N1 - Funding Information:
Dr. Brendan Marshall and Ms. Penny Roon and Ms. Libby Perry in the AU Histology and Electron Microscopy Core provided expertise with immunogold labeling and transmission electron microscopy. Dr. Paul McNeil and Mr. Tim Kurtz in the AU Cell Imaging Core assisted with confocal imaging, and Dr. Gabor Csanyi provided helpful suggestions on the Amplex Red assay. The authors are grateful to Dr. Meghan McGee-Lawrence and Michael Yu for assistance with immunostaining. Funding for this research was provided by the National Institute on Aging (P01 AG036675) and supported, in part, by the Department of Veterans Affairs (VA Merit Award 1I01CX000930-01, W.D.H.). The contents of this publication do not represent the views of the Department of Veterans Affairs or the US Government.
Publisher Copyright:
© Copyright 2017, Mary Ann Liebert, Inc. 2017.
PY - 2017/11
Y1 - 2017/11
N2 - 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 H2O2-induced oxidative stress show that H2O2 treatment of BMSCs increases the abundance of miR-183-5p in BMSC-derived EVs, and Amplex Red assays demonstrate that H2O2 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.
AB - 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 H2O2-induced oxidative stress show that H2O2 treatment of BMSCs increases the abundance of miR-183-5p in BMSC-derived EVs, and Amplex Red assays demonstrate that H2O2 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.
KW - bone
KW - exosomes
KW - miRNA
KW - oxidative stress
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UR - http://www.scopus.com/inward/citedby.url?scp=85027516950&partnerID=8YFLogxK
U2 - 10.1089/ten.tea.2016.0525
DO - 10.1089/ten.tea.2016.0525
M3 - Article
C2 - 28363268
AN - SCOPUS:85027516950
SN - 1937-3341
VL - 23
SP - 1231
EP - 1240
JO - Tissue Engineering - Part A
JF - Tissue Engineering - Part A
IS - 21-22
ER -