TY - JOUR
T1 - Expression profiling of mitochondria-associated microRNAs during osteogenic differentiation of human MSCs
AU - Zheng, Hongjun
AU - Liu, Jin
AU - Yu, Jinsheng
AU - McAlinden, Audrey
N1 - Funding Information:
This work was supported by NIH R01 AR077203 and NIH R21 AR075730 (to AM), and NIH P30 AR057235 (Washington University Musculoskeletal Research Center).We thank the Genome Technology Access Center at the McDonnell Genome Institute at Washington University School of Medicine for help with microarrays and genomic analysis. Thanks also to Erica Lantelme (FACS Core, Department of Pathology & Immunology at Washington University School of Medicine) for assistance with FACS analysis and data interpretation. We gratefully acknowledge Dr. Sanja Sviben, Gregory Strout and Dr. James Fitzpatrick for their assistance in electron microscopy studies conducted at the Washington University Center for Cellular Imaging (WUCCI), which is supported in part by Washington University School of Medicine, The Children's Discovery Institute of Washington University and St. Louis Children's Hospital (CDI-CORE-2015-505 and CDI-CORE-2019-813), the Foundation for Barnes-Jewish Hospital (3770), the Washington University Rheumatic Diseases Research Resource-based Center (NIH P30 AR073752) and the Washington University Musculoskeletal Research Center (MRC) (NIH P30 AR074992).
Funding Information:
We thank the Genome Technology Access Center at the McDonnell Genome Institute at Washington University School of Medicine for help with microarrays and genomic analysis. Thanks also to Erica Lantelme (FACS Core, Department of Pathology & Immunology at Washington University School of Medicine) for assistance with FACS analysis and data interpretation. We gratefully acknowledge Dr. Sanja Sviben, Gregory Strout and Dr. James Fitzpatrick for their assistance in electron microscopy studies conducted at the Washington University Center for Cellular Imaging (WUCCI), which is supported in part by Washington University School of Medicine , The Children's Discovery Institute of Washington University and St. Louis Children's Hospital ( CDI-CORE-2015-505 and CDI-CORE-2019-813 ), the Foundation for Barnes-Jewish Hospital ( 3770 ), the Washington University Rheumatic Diseases Research Resource-based Center (NIH P30 AR073752 ) and the Washington University Musculoskeletal Research Center (MRC) (NIH P30 AR074992 ).
Funding Information:
This work was supported by NIH R01 AR077203 and NIH R21 AR075730 (to AM), and NIH P30 AR057235 (Washington University Musculoskeletal Research Center).
Publisher Copyright:
© 2021 Elsevier Inc.
PY - 2021/10
Y1 - 2021/10
N2 - Small non-coding microRNAs (miRNAs) have the ability to target and bind to many mRNAs within the cytosol resulting in reduced protein expression and modulation of a number of cellular pathways and networks. In addition to the cytosol, miRNAs have been identified in other cellular compartments and organelles, including the mitochondria. While a few mitochondria-associated miRNAs (mitomiRs) are predicted to be derived from the mitochondrial genome, the majority appear to be transcribed from nuclear DNA and somehow transported into the mitochondria. These findings raise interesting questions about why miRNAs are located in the mitochondria and if they play a role in regulating processes within these organelles. Previously published work from our laboratory showed that miR-181a/b can regulate osteogenesis, in part, by enhancing mitochondrial metabolism. In other published studies, miR-181 paralogs and many other miRNAs have been identified in mitochondrial extracts derived from common cell lines and specific primary cells and tissues. Taken together, we were motivated to identify mitomiR expression profiles during in vitro osteogenesis. Specifically, we obtained RNA from purified mitochondrial extracts of human bone marrow-derived mesenchymal stem/stromal cells (MSCs) and from whole cell extracts of MSCs at day 0 or following osteogenic induction for 3, 7 and 14 days. Utilizing Affymetrix GeneChip™ miRNA 4.0 arrays, mitomiR expression signatures were determined at each time point. Based on the Affymetrix detection above background algorithm, the total number of miRNAs detected in MSC mitochondria extracts was 527 (non-induced MSCs), 627 (day 3 induced), 372 (day 7 induced) and 498 (day 14 induced). In addition, we identified significantly differentially-expressed mitomiRs at day 7 and day 14 of osteogenic induction when compared to day 0 (fold change ≥1.5; adjusted p value <0.05). In general, the most pronounced and highly significant changes in mitomiR expression during osteogenesis were observed at the day 7 time point. Interestingly, most miRNAs found to be differentially-expressed in mitochondria extracts did not show significantly altered expression in whole cell extracts at the same time points during osteoblast differentiation. This array study provides novel information on miRNAs associated with the mitochondria in MSCs during differentiation toward the osteoblast phenotype. These findings will guide future research to identify new miRNA candidates that may function in regulating mitochondrial function and/or bone formation, homeostasis or repair.
AB - Small non-coding microRNAs (miRNAs) have the ability to target and bind to many mRNAs within the cytosol resulting in reduced protein expression and modulation of a number of cellular pathways and networks. In addition to the cytosol, miRNAs have been identified in other cellular compartments and organelles, including the mitochondria. While a few mitochondria-associated miRNAs (mitomiRs) are predicted to be derived from the mitochondrial genome, the majority appear to be transcribed from nuclear DNA and somehow transported into the mitochondria. These findings raise interesting questions about why miRNAs are located in the mitochondria and if they play a role in regulating processes within these organelles. Previously published work from our laboratory showed that miR-181a/b can regulate osteogenesis, in part, by enhancing mitochondrial metabolism. In other published studies, miR-181 paralogs and many other miRNAs have been identified in mitochondrial extracts derived from common cell lines and specific primary cells and tissues. Taken together, we were motivated to identify mitomiR expression profiles during in vitro osteogenesis. Specifically, we obtained RNA from purified mitochondrial extracts of human bone marrow-derived mesenchymal stem/stromal cells (MSCs) and from whole cell extracts of MSCs at day 0 or following osteogenic induction for 3, 7 and 14 days. Utilizing Affymetrix GeneChip™ miRNA 4.0 arrays, mitomiR expression signatures were determined at each time point. Based on the Affymetrix detection above background algorithm, the total number of miRNAs detected in MSC mitochondria extracts was 527 (non-induced MSCs), 627 (day 3 induced), 372 (day 7 induced) and 498 (day 14 induced). In addition, we identified significantly differentially-expressed mitomiRs at day 7 and day 14 of osteogenic induction when compared to day 0 (fold change ≥1.5; adjusted p value <0.05). In general, the most pronounced and highly significant changes in mitomiR expression during osteogenesis were observed at the day 7 time point. Interestingly, most miRNAs found to be differentially-expressed in mitochondria extracts did not show significantly altered expression in whole cell extracts at the same time points during osteoblast differentiation. This array study provides novel information on miRNAs associated with the mitochondria in MSCs during differentiation toward the osteoblast phenotype. These findings will guide future research to identify new miRNA candidates that may function in regulating mitochondrial function and/or bone formation, homeostasis or repair.
KW - Bone
KW - Metabolism
KW - MicroRNA
KW - Mitochondria
KW - MitomiR
KW - Osteogenesis
UR - http://www.scopus.com/inward/record.url?scp=85108304891&partnerID=8YFLogxK
U2 - 10.1016/j.bone.2021.116058
DO - 10.1016/j.bone.2021.116058
M3 - Article
C2 - 34144232
AN - SCOPUS:85108304891
SN - 8756-3282
VL - 151
JO - Bone
JF - Bone
M1 - 116058
ER -