G protein-coupled receptor kinase 3 modulates mesenchymal stem cell proliferation and differentiation through sphingosine-1-phosphate receptor regulation
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...
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| Format: | Article |
| Language: | English |
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BMC
2022-01-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-022-02715-4 |
| _version_ | 1819281495697653760 |
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| author | Jaime M. Brozowski Roman G. Timoshchenko D. Stephen Serafin Brittney Allyn Jessica Koontz Emily M. Rabjohns Rishi R. Rampersad Yinshi Ren Amanda M. Eudy Taylor F. Harris David Abraham Daniel Mattox Clinton T. Rubin Matthew J. Hilton Janet Rubin Nancy L. Allbritton Matthew J. Billard Teresa K. Tarrant |
| author_facet | Jaime M. Brozowski Roman G. Timoshchenko D. Stephen Serafin Brittney Allyn Jessica Koontz Emily M. Rabjohns Rishi R. Rampersad Yinshi Ren Amanda M. Eudy Taylor F. Harris David Abraham Daniel Mattox Clinton T. Rubin Matthew J. Hilton Janet Rubin Nancy L. Allbritton Matthew J. Billard Teresa K. Tarrant |
| author_sort | Jaime M. Brozowski |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-24T01:00:36Z |
| format | Article |
| id | doaj.art-124f702e78d146ed89416332f38c7248 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-24T01:00:36Z |
| publishDate | 2022-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-124f702e78d146ed89416332f38c72482022-12-21T17:23:22ZengBMCStem Cell Research & Therapy1757-65122022-01-0113111510.1186/s13287-022-02715-4G protein-coupled receptor kinase 3 modulates mesenchymal stem cell proliferation and differentiation through sphingosine-1-phosphate receptor regulationJaime M. Brozowski0Roman G. Timoshchenko1D. Stephen Serafin2Brittney Allyn3Jessica Koontz4Emily M. Rabjohns5Rishi R. Rampersad6Yinshi Ren7Amanda M. Eudy8Taylor F. Harris9David Abraham10Daniel Mattox11Clinton T. Rubin12Matthew J. Hilton13Janet Rubin14Nancy L. Allbritton15Matthew J. Billard16Teresa K. Tarrant17Department of Microbiology and Immunology, University of North Carolina at Chapel HillThurston Arthritis Research Center, University of North Carolina at Chapel HillThurston Arthritis Research Center, University of North Carolina at Chapel HillThurston Arthritis Research Center, University of North Carolina at Chapel HillThurston Arthritis Research Center, University of North Carolina at Chapel HillDepartment of Medicine, Division of Rheumatology and Immunology, Duke UniversityDepartment of Medicine, Division of Rheumatology and Immunology, Duke UniversityDepartment of Orthopaedic Surgery, Duke Orthopaedic Cellular, Developmental and Genome Laboratories, Duke University School of MedicineDepartment of Medicine, Division of Rheumatology and Immunology, Duke UniversityDepartment of Chemistry, University of North Carolina at Chapel HillDepartment of Chemistry, University of North Carolina at Chapel HillThurston Arthritis Research Center, University of North Carolina at Chapel HillDepartment of Biomedical Engineering at Stony, Brook UniversityDepartment of Orthopaedic Surgery, Duke Orthopaedic Cellular, Developmental and Genome Laboratories, Duke University School of MedicineDepartment of Medicine, University of North Carolina at Chapel HillDepartment of Chemistry, University of North Carolina at Chapel HillThurston Arthritis Research Center, University of North Carolina at Chapel HillDepartment of Microbiology and Immunology, University of North Carolina at Chapel HillAbstract 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.https://doi.org/10.1186/s13287-022-02715-4Bone marrow-derived mesenchymal stem cellsMesenchymal stem cellsG protein-coupled receptors (GPCRs)G protein-coupled receptor kinase 3 (GRK3)Sphingosine-1-phosphate (S1P)Sphingosine-1-phosphate receptor (S1PR) |
| spellingShingle | Jaime M. Brozowski Roman G. Timoshchenko D. Stephen Serafin Brittney Allyn Jessica Koontz Emily M. Rabjohns Rishi R. Rampersad Yinshi Ren Amanda M. Eudy Taylor F. Harris David Abraham Daniel Mattox Clinton T. Rubin Matthew J. Hilton Janet Rubin Nancy L. Allbritton Matthew J. Billard Teresa K. Tarrant G protein-coupled receptor kinase 3 modulates mesenchymal stem cell proliferation and differentiation through sphingosine-1-phosphate receptor regulation Stem Cell Research & Therapy Bone marrow-derived mesenchymal stem cells Mesenchymal stem cells G protein-coupled receptors (GPCRs) G protein-coupled receptor kinase 3 (GRK3) Sphingosine-1-phosphate (S1P) Sphingosine-1-phosphate receptor (S1PR) |
| title | G protein-coupled receptor kinase 3 modulates mesenchymal stem cell proliferation and differentiation through sphingosine-1-phosphate receptor regulation |
| title_full | G protein-coupled receptor kinase 3 modulates mesenchymal stem cell proliferation and differentiation through sphingosine-1-phosphate receptor regulation |
| title_fullStr | G protein-coupled receptor kinase 3 modulates mesenchymal stem cell proliferation and differentiation through sphingosine-1-phosphate receptor regulation |
| title_full_unstemmed | G protein-coupled receptor kinase 3 modulates mesenchymal stem cell proliferation and differentiation through sphingosine-1-phosphate receptor regulation |
| title_short | G protein-coupled receptor kinase 3 modulates mesenchymal stem cell proliferation and differentiation through sphingosine-1-phosphate receptor regulation |
| title_sort | g protein coupled receptor kinase 3 modulates mesenchymal stem cell proliferation and differentiation through sphingosine 1 phosphate receptor regulation |
| topic | Bone marrow-derived mesenchymal stem cells Mesenchymal stem cells G protein-coupled receptors (GPCRs) G protein-coupled receptor kinase 3 (GRK3) Sphingosine-1-phosphate (S1P) Sphingosine-1-phosphate receptor (S1PR) |
| url | https://doi.org/10.1186/s13287-022-02715-4 |
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