title: 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 H2O2-induced oxidative stress show that H2O2treatment of BMSCs increases the abundance of miR-183-5p in BMSC-derived EVs, and Amplex Red assays demonstrate that H2O2is 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.
authors.affiliation: Department of Cellular Biology & Anatomy, Medical College of Georgia, Augusta University , Augusta, Georgia .
authors.forename: Colleen
authors.initials: C
authors.lastname: Davis
authors.affiliation: Department of Cellular Biology & Anatomy, Medical College of Georgia, Augusta University , Augusta, Georgia .
authors.forename: Amy
authors.initials: A
authors.lastname: Dukes
authors.affiliation: Department of Cellular Biology & Anatomy, Medical College of Georgia, Augusta University , Augusta, Georgia .
authors.forename: Michelle
authors.initials: M
authors.lastname: Drewry
authors.affiliation: Department of Cellular Biology & Anatomy, Medical College of Georgia, Augusta University , Augusta, Georgia .
authors.forename: Inas
authors.initials: I
authors.lastname: Helwa
authors.affiliation: Department of Cellular Biology & Anatomy, Medical College of Georgia, Augusta University , Augusta, Georgia .
authors.forename: Maribeth H
authors.initials: MH
authors.lastname: Johnson
authors.affiliation: Department of Cellular Biology & Anatomy, Medical College of Georgia, Augusta University , Augusta, Georgia .
authors.forename: Carlos M
authors.initials: CM
authors.lastname: Isales
authors.affiliation: Department of Cellular Biology & Anatomy, Medical College of Georgia, Augusta University , Augusta, Georgia .
authors.forename: William D
authors.initials: WD
authors.lastname: Hill
authors.affiliation: Department of Cellular Biology & Anatomy, Medical College of Georgia, Augusta University , Augusta, Georgia .
authors.forename: Yutao
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Cellular Biology & Anatomy, Medical College of Georgia, Augusta University , Augusta, Georgia .
authors.forename: Xingming
authors.initials: X
authors.lastname: Shi
authors.affiliation: Department of Cellular Biology & Anatomy, Medical College of Georgia, Augusta University , Augusta, Georgia .
authors.forename: Sadanand
authors.initials: S
authors.lastname: Fulzele
authors.affiliation: Department of Cellular Biology & Anatomy, Medical College of Georgia, Augusta University , Augusta, Georgia .
authors.forename: Mark W
authors.initials: MW
authors.lastname: Hamrick
mesh_terms: D000375:Aging / Q000235:genetics*; D000818:Animals; D001842:Bone and Bones / Q000166:cytology / Q000378:metabolism*; D002113:Calcification, Physiologic / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics; D049109:Cell Proliferation; D016922:Cellular Senescence; D004705:Endocytosis / Q000235:genetics; D000067128:Extracellular Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D020869:Gene Expression Profiling; D051547:Heme Oxygenase-1 / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D010012:Osteogenesis / Q000235:genetics; D018384:Oxidative Stress; D014162:Transfection
chemical_list: D035683:MicroRNAs; C543802:Mirn183 microRNA, mouse; D051547:Heme Oxygenase-1
keywords: bone; exosomes; miRNA; oxidative stress