MiRNAs Expression Profiling in Raw264.7 Macrophages after Nfatc1-Knockdown Elucidates Potential Pathways Involved in Osteoclasts Differentiation

Differentiation of macrophages toward osteoclasts is crucial for bone homeostasis but can be detrimental in disease states, including osteoporosis and cancer. Therefore, understanding the osteoclast differentiation process and the underlying regulatory mechanisms may facilitate the identification of...

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Autores principales:	Roberta Russo, Francesca Zito, Nadia Lampiasi
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	osteoclastogenesis differential expression MAPK pathway NFATc1 PCR arrays siRNA transfection Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/7621d03a9792427ab1bfbf1de5eb483c
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spelling	oai:doaj.org-article:7621d03a9792427ab1bfbf1de5eb483c2021-11-25T16:46:49ZMiRNAs Expression Profiling in Raw264.7 Macrophages after Nfatc1-Knockdown Elucidates Potential Pathways Involved in Osteoclasts Differentiation10.3390/biology101110802079-7737https://doaj.org/article/7621d03a9792427ab1bfbf1de5eb483c2021-10-01T00:00:00Zhttps://www.mdpi.com/2079-7737/10/11/1080https://doaj.org/toc/2079-7737Differentiation of macrophages toward osteoclasts is crucial for bone homeostasis but can be detrimental in disease states, including osteoporosis and cancer. Therefore, understanding the osteoclast differentiation process and the underlying regulatory mechanisms may facilitate the identification of new therapeutic targets. Hereby, we tried to reveal new miRNAs potentially involved in the regulation of early steps of osteoclastogenesis, with a particular focus on those possibly correlated with NFATc1 expression, by studying miRNAs profiling. During the first 24 h of osteoclastogenesis, 38 miRNAs were differentially expressed between undifferentiated and RANKL-stimulated RAW264.7 cells, while 10 miRNAs were differentially expressed between RANKL-stimulated cells transfected with negative control or NFATc1-siRNAs. Among others, the expression levels of miR-411, miR-144 and members of miR-29, miR-30, and miR-23 families changed after RANKL stimulation. Moreover, the potential role of miR-124 during osteoclastogenesis was explored by transient cell transfection with anti-miR-124 or miR-124-mimic. Two relatively unknown miRNAs, miR-880-3p and miR-295-3p, were differentially expressed between RANKL-stimulated/wild-type and RANKL-stimulated/NFATc1-silenced cells, suggesting their possible correlation with NFATc1. KEGG enrichment analyses showed that kinase and phosphatase enzymes were among the predicted targets for many of the studied miRNAs. In conclusion, our study provides new data on the potential role and possible targets of new miRNAs during osteoclastogenesis.Roberta RussoFrancesca ZitoNadia LampiasiMDPI AGarticleosteoclastogenesisdifferential expressionMAPK pathwayNFATc1PCR arrayssiRNA transfectionBiology (General)QH301-705.5ENBiology, Vol 10, Iss 1080, p 1080 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	osteoclastogenesis differential expression MAPK pathway NFATc1 PCR arrays siRNA transfection Biology (General) QH301-705.5
spellingShingle	osteoclastogenesis differential expression MAPK pathway NFATc1 PCR arrays siRNA transfection Biology (General) QH301-705.5 Roberta Russo Francesca Zito Nadia Lampiasi MiRNAs Expression Profiling in Raw264.7 Macrophages after Nfatc1-Knockdown Elucidates Potential Pathways Involved in Osteoclasts Differentiation
description	Differentiation of macrophages toward osteoclasts is crucial for bone homeostasis but can be detrimental in disease states, including osteoporosis and cancer. Therefore, understanding the osteoclast differentiation process and the underlying regulatory mechanisms may facilitate the identification of new therapeutic targets. Hereby, we tried to reveal new miRNAs potentially involved in the regulation of early steps of osteoclastogenesis, with a particular focus on those possibly correlated with NFATc1 expression, by studying miRNAs profiling. During the first 24 h of osteoclastogenesis, 38 miRNAs were differentially expressed between undifferentiated and RANKL-stimulated RAW264.7 cells, while 10 miRNAs were differentially expressed between RANKL-stimulated cells transfected with negative control or NFATc1-siRNAs. Among others, the expression levels of miR-411, miR-144 and members of miR-29, miR-30, and miR-23 families changed after RANKL stimulation. Moreover, the potential role of miR-124 during osteoclastogenesis was explored by transient cell transfection with anti-miR-124 or miR-124-mimic. Two relatively unknown miRNAs, miR-880-3p and miR-295-3p, were differentially expressed between RANKL-stimulated/wild-type and RANKL-stimulated/NFATc1-silenced cells, suggesting their possible correlation with NFATc1. KEGG enrichment analyses showed that kinase and phosphatase enzymes were among the predicted targets for many of the studied miRNAs. In conclusion, our study provides new data on the potential role and possible targets of new miRNAs during osteoclastogenesis.
format	article
author	Roberta Russo Francesca Zito Nadia Lampiasi
author_facet	Roberta Russo Francesca Zito Nadia Lampiasi
author_sort	Roberta Russo
title	MiRNAs Expression Profiling in Raw264.7 Macrophages after Nfatc1-Knockdown Elucidates Potential Pathways Involved in Osteoclasts Differentiation
title_short	MiRNAs Expression Profiling in Raw264.7 Macrophages after Nfatc1-Knockdown Elucidates Potential Pathways Involved in Osteoclasts Differentiation
title_full	MiRNAs Expression Profiling in Raw264.7 Macrophages after Nfatc1-Knockdown Elucidates Potential Pathways Involved in Osteoclasts Differentiation
title_fullStr	MiRNAs Expression Profiling in Raw264.7 Macrophages after Nfatc1-Knockdown Elucidates Potential Pathways Involved in Osteoclasts Differentiation
title_full_unstemmed	MiRNAs Expression Profiling in Raw264.7 Macrophages after Nfatc1-Knockdown Elucidates Potential Pathways Involved in Osteoclasts Differentiation
title_sort	mirnas expression profiling in raw264.7 macrophages after nfatc1-knockdown elucidates potential pathways involved in osteoclasts differentiation
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/7621d03a9792427ab1bfbf1de5eb483c
work_keys_str_mv	AT robertarusso mirnasexpressionprofilinginraw2647macrophagesafternfatc1knockdownelucidatespotentialpathwaysinvolvedinosteoclastsdifferentiation AT francescazito mirnasexpressionprofilinginraw2647macrophagesafternfatc1knockdownelucidatespotentialpathwaysinvolvedinosteoclastsdifferentiation AT nadialampiasi mirnasexpressionprofilinginraw2647macrophagesafternfatc1knockdownelucidatespotentialpathwaysinvolvedinosteoclastsdifferentiation
_version_	1718412962758131712

MiRNAs Expression Profiling in Raw264.7 Macrophages after Nfatc1-Knockdown Elucidates Potential Pathways Involved in Osteoclasts Differentiation

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