Cargando…

Pathway Analysis of MicroRNA Expression Profile during Murine Osteoclastogenesis

To design novel therapeutics against bone loss, understanding the molecular mechanisms regulating osteoclastogenesis is critical. Osteoclast formation and function are tightly regulated by transcriptional, post-transcriptional and post-translational mechanisms. This stringent regulation is crucial t...

Descripción completa

Detalles Bibliográficos
Autores principales: Franceschetti, Tiziana, Dole, Neha S., Kessler, Catherine B., Lee, Sun-Kyeong, Delany, Anne M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4164525/
https://www.ncbi.nlm.nih.gov/pubmed/25222202
http://dx.doi.org/10.1371/journal.pone.0107262
_version_ 1782334967498407936
author Franceschetti, Tiziana
Dole, Neha S.
Kessler, Catherine B.
Lee, Sun-Kyeong
Delany, Anne M.
author_facet Franceschetti, Tiziana
Dole, Neha S.
Kessler, Catherine B.
Lee, Sun-Kyeong
Delany, Anne M.
author_sort Franceschetti, Tiziana
collection PubMed
description To design novel therapeutics against bone loss, understanding the molecular mechanisms regulating osteoclastogenesis is critical. Osteoclast formation and function are tightly regulated by transcriptional, post-transcriptional and post-translational mechanisms. This stringent regulation is crucial to prevent excessive or insufficient bone resorption and to maintain bone homeostasis. microRNAs (miRNAs) are key post-transcriptional regulators that repress expression of target mRNAs controlling osteoclast proliferation, differentiation, and apoptosis. Disruption of miRNA-mediated regulation alters osteoclast formation and bone resorption. Prior studies profiled miRNA expression in murine osteoclast precursors treated with RANKL for 24 hours. However, a more complete miRNA signature, encompassing early, mid and late stages of osteoclastogenesis, is wanting. An Agilent microarray platform was used to analyze expression of mature miRNAs in an enriched population of murine bone marrow osteoclast precursors (depleted of B220(+) and CD3(+) cells) undergoing 1, 3, or 5 days of RANKL-driven differentiation. Expression of 93 miRNAs, changed by >2 fold during early, mid, and late stages of osteoclastogenesis, were identified and sorted into 7 clusters. We validated the function and expression of miR-365, miR-451, and miR-99b, which were found in distinct clusters. Inhibition of miR-365 increased osteoclast number but decreased osteoclast size, while miR-99b inhibition decreased both osteoclast number and size. In contrast, overexpression of miR-451 had no effect. Computational analyses predicted mTOR, PI3 kinase/AKT, cell-matrix interactions, actin cytoskeleton organization, focal adhesion, and axon guidance pathways to be top targets of several miRNA clusters. This suggests that many miRNA clusters differentially expressed during osteoclastogenesis converge on some key functional pathways. Overall, our study is unique in that we identified miRNAs differentially expressed during early, mid, and late osteoclastogenesis in a population of primary mouse bone marrow cells enriched for osteoclast progenitors. This novel data set contributes to our understanding of the molecular mechanisms regulating the complex process of osteoclast differentiation.
format Online
Article
Text
id pubmed-4164525
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-41645252014-09-19 Pathway Analysis of MicroRNA Expression Profile during Murine Osteoclastogenesis Franceschetti, Tiziana Dole, Neha S. Kessler, Catherine B. Lee, Sun-Kyeong Delany, Anne M. PLoS One Research Article To design novel therapeutics against bone loss, understanding the molecular mechanisms regulating osteoclastogenesis is critical. Osteoclast formation and function are tightly regulated by transcriptional, post-transcriptional and post-translational mechanisms. This stringent regulation is crucial to prevent excessive or insufficient bone resorption and to maintain bone homeostasis. microRNAs (miRNAs) are key post-transcriptional regulators that repress expression of target mRNAs controlling osteoclast proliferation, differentiation, and apoptosis. Disruption of miRNA-mediated regulation alters osteoclast formation and bone resorption. Prior studies profiled miRNA expression in murine osteoclast precursors treated with RANKL for 24 hours. However, a more complete miRNA signature, encompassing early, mid and late stages of osteoclastogenesis, is wanting. An Agilent microarray platform was used to analyze expression of mature miRNAs in an enriched population of murine bone marrow osteoclast precursors (depleted of B220(+) and CD3(+) cells) undergoing 1, 3, or 5 days of RANKL-driven differentiation. Expression of 93 miRNAs, changed by >2 fold during early, mid, and late stages of osteoclastogenesis, were identified and sorted into 7 clusters. We validated the function and expression of miR-365, miR-451, and miR-99b, which were found in distinct clusters. Inhibition of miR-365 increased osteoclast number but decreased osteoclast size, while miR-99b inhibition decreased both osteoclast number and size. In contrast, overexpression of miR-451 had no effect. Computational analyses predicted mTOR, PI3 kinase/AKT, cell-matrix interactions, actin cytoskeleton organization, focal adhesion, and axon guidance pathways to be top targets of several miRNA clusters. This suggests that many miRNA clusters differentially expressed during osteoclastogenesis converge on some key functional pathways. Overall, our study is unique in that we identified miRNAs differentially expressed during early, mid, and late osteoclastogenesis in a population of primary mouse bone marrow cells enriched for osteoclast progenitors. This novel data set contributes to our understanding of the molecular mechanisms regulating the complex process of osteoclast differentiation. Public Library of Science 2014-09-15 /pmc/articles/PMC4164525/ /pubmed/25222202 http://dx.doi.org/10.1371/journal.pone.0107262 Text en © 2014 Franceschetti et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Franceschetti, Tiziana
Dole, Neha S.
Kessler, Catherine B.
Lee, Sun-Kyeong
Delany, Anne M.
Pathway Analysis of MicroRNA Expression Profile during Murine Osteoclastogenesis
title Pathway Analysis of MicroRNA Expression Profile during Murine Osteoclastogenesis
title_full Pathway Analysis of MicroRNA Expression Profile during Murine Osteoclastogenesis
title_fullStr Pathway Analysis of MicroRNA Expression Profile during Murine Osteoclastogenesis
title_full_unstemmed Pathway Analysis of MicroRNA Expression Profile during Murine Osteoclastogenesis
title_short Pathway Analysis of MicroRNA Expression Profile during Murine Osteoclastogenesis
title_sort pathway analysis of microrna expression profile during murine osteoclastogenesis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4164525/
https://www.ncbi.nlm.nih.gov/pubmed/25222202
http://dx.doi.org/10.1371/journal.pone.0107262
work_keys_str_mv AT franceschettitiziana pathwayanalysisofmicrornaexpressionprofileduringmurineosteoclastogenesis
AT dolenehas pathwayanalysisofmicrornaexpressionprofileduringmurineosteoclastogenesis
AT kesslercatherineb pathwayanalysisofmicrornaexpressionprofileduringmurineosteoclastogenesis
AT leesunkyeong pathwayanalysisofmicrornaexpressionprofileduringmurineosteoclastogenesis
AT delanyannem pathwayanalysisofmicrornaexpressionprofileduringmurineosteoclastogenesis