G protein-coupled receptor kinase 3 modulates mesenchymal stem cell proliferation and differentiation through sphingosine-1-phosphate receptor regulation.

Brozowski, Jaime M; Timoshchenko, Roman G; Serafin, D Stephen; Allyn, Brittney; Koontz, Jessica; Rabjohns, Emily M; Rampersad, Rishi R; Ren, Yinshi; Eudy, Amanda M; Harris, Taylor F; Abraham, David; Mattox, Daniel; Rubin, Clinton T; Hilton, Matthew J; Rubin, Janet; Allbritton, Nancy L; Billard, Matthew J; Tarrant, Teresa K

G protein-coupled receptor kinase 3 modulates mesenchymal stem cell proliferation and differentiation through sphingosine-1-phosphate receptor regulation.

dc.contributor.author	Brozowski, Jaime M
dc.contributor.author	Timoshchenko, Roman G
dc.contributor.author	Serafin, D Stephen
dc.contributor.author	Allyn, Brittney
dc.contributor.author	Koontz, Jessica
dc.contributor.author	Rabjohns, Emily M
dc.contributor.author	Rampersad, Rishi R
dc.contributor.author	Ren, Yinshi
dc.contributor.author	Eudy, Amanda M
dc.contributor.author	Harris, Taylor F
dc.contributor.author	Abraham, David
dc.contributor.author	Mattox, Daniel
dc.contributor.author	Rubin, Clinton T
dc.contributor.author	Hilton, Matthew J
dc.contributor.author	Rubin, Janet
dc.contributor.author	Allbritton, Nancy L
dc.contributor.author	Billard, Matthew J
dc.contributor.author	Tarrant, Teresa K
dc.date.accessioned	2022-04-21T18:59:01Z
dc.date.available	2022-04-21T18:59:01Z
dc.date.issued	2022-01-29
dc.date.updated	2022-04-21T18:59:01Z
dc.description.abstract	Background The bone marrow niche supports hematopoietic cell development through intimate contact with multipotent stromal mesenchymal stem cells; however, the intracellular signaling, function, and regulation of such supportive niche cells are still being defined. Our study was designed to understand how G protein receptor kinase 3 (GRK3) affects bone marrow mesenchymal stem cell function by examining primary cells from GRK3-deficient mice, which we have previously published to have a hypercellular bone marrow and leukocytosis through negative regulation of CXCL12/CXCR4 signaling. Methods Murine GRK3-deficient bone marrow mesenchymal stromal cells were harvested and cultured to differentiate into three lineages (adipocyte, chondrocyte, and osteoblast) to confirm multipotency and compared to wild type cells. Immunoblotting, modified-TANGO experiments, and flow cytometry were used to further examine the effects of GRK3 deficiency on bone marrow mesenchymal stromal cell receptor signaling. Microcomputed tomography was used to determine trabecular and cortical bone composition of GRK3-deficient mice and standard ELISA to quantitate CXCL12 production from cellular cultures. Results GRK3-deficient, bone marrow-derived mesenchymal stem cells exhibit enhanced and earlier osteogenic differentiation in vitro. The addition of a sphingosine kinase inhibitor abrogated the osteogenic proliferation and differentiation, suggesting that sphingosine-1-phosphate receptor signaling was a putative G protein-coupled receptor regulated by GRK3. Immunoblotting showed prolonged ERK1/2 signaling after stimulation with sphingosine-1-phosphate in GRK3-deficient cells, and modified-TANGO assays suggested the involvement of β-arrestin-2 in sphingosine-1-phosphate receptor internalization. Conclusions Our work suggests that GRK3 regulates sphingosine-1-phosphate receptor signaling on bone marrow mesenchymal stem cells by recruiting β-arrestin to the occupied GPCR to promote internalization, and lack of such regulation affects mesenchymal stem cell functionality.
dc.identifier.issn	1757-6512
dc.identifier.issn	1757-6512
dc.identifier.uri	https://hdl.handle.net/10161/24859
dc.publisher	Springer Science and Business Media LLC
dc.relation.ispartof	Stem cell research & therapy
dc.relation.isversionof	10.1186/s13287-022-02715-4
dc.subject	Mesenchymal Stem Cells
dc.subject	Animals
dc.subject	Mice
dc.subject	Cell Differentiation
dc.subject	Cell Proliferation
dc.subject	Osteogenesis
dc.subject	X-Ray Microtomography
dc.subject	Sphingosine-1-Phosphate Receptors
dc.title	G protein-coupled receptor kinase 3 modulates mesenchymal stem cell proliferation and differentiation through sphingosine-1-phosphate receptor regulation.
dc.type	Conference
duke.contributor.orcid	Eudy, Amanda M\|0000-0002-3107-5545
duke.contributor.orcid	Hilton, Matthew J\|0000-0003-3165-267X
duke.contributor.orcid	Tarrant, Teresa K\|0000-0003-4067-5363
pubs.begin-page	37
pubs.issue	1
pubs.organisational-group	Duke
pubs.organisational-group	School of Medicine
pubs.organisational-group	Basic Science Departments
pubs.organisational-group	Clinical Science Departments
pubs.organisational-group	Cell Biology
pubs.organisational-group	Medicine
pubs.organisational-group	Orthopaedics
pubs.organisational-group	Medicine, Rheumatology and Immunology
pubs.publication-status	Published
pubs.volume	13