Inhibition of Sphingosine-1-Phosphate Receptor 2 by JTE013 Promoted Osteogenesis by Increasing Vesicle Trafficking, Wnt/Ca<sup>2+</sup>, and BMP/Smad Signaling

Sphingosine-1-phosphate receptor 2 (S1PR2) is a G protein-coupled receptor that regulates various immune responses. Herein, we determine the effects of a S1PR2 antagonist (JTE013) or a S1PR2 shRNA on osteogenesis by culturing murine bone marrow stromal cells (BMSCs) in osteogenic media with JTE013,...

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Autores principales:	Simon Lin, Subramanya Pandruvada, Hong Yu
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	S1PR2 JTE013 osteogenesis matrix vesicle vesicle trafficking Wnt Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/28f43dc6f0ea46f0a3b480db16276085
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spelling	oai:doaj.org-article:28f43dc6f0ea46f0a3b480db162760852021-11-11T17:27:24ZInhibition of Sphingosine-1-Phosphate Receptor 2 by JTE013 Promoted Osteogenesis by Increasing Vesicle Trafficking, Wnt/Ca<sup>2+</sup>, and BMP/Smad Signaling10.3390/ijms2221120601422-00671661-6596https://doaj.org/article/28f43dc6f0ea46f0a3b480db162760852021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/12060https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Sphingosine-1-phosphate receptor 2 (S1PR2) is a G protein-coupled receptor that regulates various immune responses. Herein, we determine the effects of a S1PR2 antagonist (JTE013) or a S1PR2 shRNA on osteogenesis by culturing murine bone marrow stromal cells (BMSCs) in osteogenic media with JTE013, dimethylsulfoxide (DMSO), a S1PR2 shRNA, or a control shRNA. Treatment with JTE013 or the S1PR2 shRNA increased alkaline phosphatase and alizarin red s staining, and enhanced alkaline phosphatase, RUNX2, osteocalcin, and osterix mRNA levels in BMSCs compared with the controls. Protein analysis revealed that a high dose of JTE013 (4 or 8 μM) increased vesicle trafficking-associated proteins (F-actin, clathrin, Early Endosome Antigen 1 (EEA1), and syntaxin 6) and Wnt/Ca2+ signaling. On the other hand, a low dose of JTE013 (1 to 2 μM) increased BMP/Smad signaling. In contrast, the S1PR2 shRNA reduced vesicle trafficking-associated proteins and attenuated Wnts and BMP/Smad signaling, but enhanced p-CaMKII compared with the control, suggesting that the S1PR2 shRNA influenced osteogenesis via different signaling pathways. Moreover, inhibiting protein trafficking by brefeldin A in BMSCs suppressed Wnts and BMPRs expressions. These data supported that enhanced osteogenesis in JTE013-treated BMSCs is associated with increased vesicle trafficking, which promotes the synthesis and transport of osteogenic protein and matrix vesicles and enhances matrix mineralization.Simon LinSubramanya PandruvadaHong YuMDPI AGarticleS1PR2JTE013osteogenesismatrix vesiclevesicle traffickingWntBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12060, p 12060 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	S1PR2 JTE013 osteogenesis matrix vesicle vesicle trafficking Wnt Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	S1PR2 JTE013 osteogenesis matrix vesicle vesicle trafficking Wnt Biology (General) QH301-705.5 Chemistry QD1-999 Simon Lin Subramanya Pandruvada Hong Yu Inhibition of Sphingosine-1-Phosphate Receptor 2 by JTE013 Promoted Osteogenesis by Increasing Vesicle Trafficking, Wnt/Ca<sup>2+</sup>, and BMP/Smad Signaling
description	Sphingosine-1-phosphate receptor 2 (S1PR2) is a G protein-coupled receptor that regulates various immune responses. Herein, we determine the effects of a S1PR2 antagonist (JTE013) or a S1PR2 shRNA on osteogenesis by culturing murine bone marrow stromal cells (BMSCs) in osteogenic media with JTE013, dimethylsulfoxide (DMSO), a S1PR2 shRNA, or a control shRNA. Treatment with JTE013 or the S1PR2 shRNA increased alkaline phosphatase and alizarin red s staining, and enhanced alkaline phosphatase, RUNX2, osteocalcin, and osterix mRNA levels in BMSCs compared with the controls. Protein analysis revealed that a high dose of JTE013 (4 or 8 μM) increased vesicle trafficking-associated proteins (F-actin, clathrin, Early Endosome Antigen 1 (EEA1), and syntaxin 6) and Wnt/Ca2+ signaling. On the other hand, a low dose of JTE013 (1 to 2 μM) increased BMP/Smad signaling. In contrast, the S1PR2 shRNA reduced vesicle trafficking-associated proteins and attenuated Wnts and BMP/Smad signaling, but enhanced p-CaMKII compared with the control, suggesting that the S1PR2 shRNA influenced osteogenesis via different signaling pathways. Moreover, inhibiting protein trafficking by brefeldin A in BMSCs suppressed Wnts and BMPRs expressions. These data supported that enhanced osteogenesis in JTE013-treated BMSCs is associated with increased vesicle trafficking, which promotes the synthesis and transport of osteogenic protein and matrix vesicles and enhances matrix mineralization.
format	article
author	Simon Lin Subramanya Pandruvada Hong Yu
author_facet	Simon Lin Subramanya Pandruvada Hong Yu
author_sort	Simon Lin
title	Inhibition of Sphingosine-1-Phosphate Receptor 2 by JTE013 Promoted Osteogenesis by Increasing Vesicle Trafficking, Wnt/Ca<sup>2+</sup>, and BMP/Smad Signaling
title_short	Inhibition of Sphingosine-1-Phosphate Receptor 2 by JTE013 Promoted Osteogenesis by Increasing Vesicle Trafficking, Wnt/Ca<sup>2+</sup>, and BMP/Smad Signaling
title_full	Inhibition of Sphingosine-1-Phosphate Receptor 2 by JTE013 Promoted Osteogenesis by Increasing Vesicle Trafficking, Wnt/Ca<sup>2+</sup>, and BMP/Smad Signaling
title_fullStr	Inhibition of Sphingosine-1-Phosphate Receptor 2 by JTE013 Promoted Osteogenesis by Increasing Vesicle Trafficking, Wnt/Ca<sup>2+</sup>, and BMP/Smad Signaling
title_full_unstemmed	Inhibition of Sphingosine-1-Phosphate Receptor 2 by JTE013 Promoted Osteogenesis by Increasing Vesicle Trafficking, Wnt/Ca<sup>2+</sup>, and BMP/Smad Signaling
title_sort	inhibition of sphingosine-1-phosphate receptor 2 by jte013 promoted osteogenesis by increasing vesicle trafficking, wnt/ca<sup>2+</sup>, and bmp/smad signaling
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/28f43dc6f0ea46f0a3b480db16276085
work_keys_str_mv	AT simonlin inhibitionofsphingosine1phosphatereceptor2byjte013promotedosteogenesisbyincreasingvesicletraffickingwntcasup2supandbmpsmadsignaling AT subramanyapandruvada inhibitionofsphingosine1phosphatereceptor2byjte013promotedosteogenesisbyincreasingvesicletraffickingwntcasup2supandbmpsmadsignaling AT hongyu inhibitionofsphingosine1phosphatereceptor2byjte013promotedosteogenesisbyincreasingvesicletraffickingwntcasup2supandbmpsmadsignaling
_version_	1718432052001374208

Inhibition of Sphingosine-1-Phosphate Receptor 2 by JTE013 Promoted Osteogenesis by Increasing Vesicle Trafficking, Wnt/Ca<sup>2+</sup>, and BMP/Smad Signaling

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