Vascular Endothelial Growth Factor (VEGF) and Platelet (PF-4) Factor 4 Inputs Modulate Human Microvascular Endothelial Signaling in a Three-Dimensional Matrix Migration Context
The process of angiogenesis is under complex regulation in adult organisms, particularly as it often occurs in an inflammatory post-wound environment. As such, there are many impacting factors that will regulate the generation of new blood vessels which include not only pro-angiogenic growth factors...
| Main Authors: | , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
American Society for Biochemistry and Molecular Biology (ASBMB)
2014
|
| Online Access: | http://hdl.handle.net/1721.1/89149 https://orcid.org/0000-0002-1545-1651 https://orcid.org/0000-0002-7232-304X https://orcid.org/0000-0002-3856-7454 |
| _version_ | 1826214821099470848 |
|---|---|
| author | Hang, Ta-Chun Tedford, Nathan C. Reddy, Raven J. Rimchala, Tharathorn Wells, Alan White, Forest M. Kamm, Roger Dale Lauffenburger, Douglas A. |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Hang, Ta-Chun Tedford, Nathan C. Reddy, Raven J. Rimchala, Tharathorn Wells, Alan White, Forest M. Kamm, Roger Dale Lauffenburger, Douglas A. |
| author_sort | Hang, Ta-Chun |
| collection | MIT |
| description | The process of angiogenesis is under complex regulation in adult organisms, particularly as it often occurs in an inflammatory post-wound environment. As such, there are many impacting factors that will regulate the generation of new blood vessels which include not only pro-angiogenic growth factors such as vascular endothelial growth factor, but also angiostatic factors. During initial postwound hemostasis, a large initial bolus of platelet factor 4 is released into localized areas of damage before progression of wound healing toward tissue homeostasis. Because of its early presence and high concentration, the angiostatic chemokine platelet factor 4, which can induce endothelial anoikis, can strongly affect angiogenesis. In our work, we explored signaling crosstalk interactions between vascular endothelial growth factor and platelet factor 4 using phosphotyrosine-enriched mass spectrometry methods on human dermal microvascular endothelial cells cultured under conditions facilitating migratory sprouting into collagen gel matrices. We developed new methods to enable mass spectrometry-based phosphorylation analysis of primary cells cultured on collagen gels, and quantified signaling pathways over the first 48 h of treatment with vascular endothelial growth factor in the presence or absence of platelet factor 4. By observing early and late signaling dynamics in tandem with correlation network modeling, we found that platelet factor 4 has significant crosstalk with vascular endothelial growth factor by modulating cell migration and polarization pathways, centered around P38α MAPK, Src family kinases Fyn and Lyn, along with FAK. Interestingly, we found EphA2 correlational topology to strongly involve key migration-related signaling nodes after introduction of platelet factor 4, indicating an influence of the angiostatic factor on this ambiguous but generally angiogenic signal in this complex environment. |
| first_indexed | 2024-09-23T16:11:41Z |
| format | Article |
| id | mit-1721.1/89149 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:11:41Z |
| publishDate | 2014 |
| publisher | American Society for Biochemistry and Molecular Biology (ASBMB) |
| record_format | dspace |
| spelling | mit-1721.1/891492022-10-02T06:57:55Z Vascular Endothelial Growth Factor (VEGF) and Platelet (PF-4) Factor 4 Inputs Modulate Human Microvascular Endothelial Signaling in a Three-Dimensional Matrix Migration Context Hang, Ta-Chun Tedford, Nathan C. Reddy, Raven J. Rimchala, Tharathorn Wells, Alan White, Forest M. Kamm, Roger Dale Lauffenburger, Douglas A. Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Mechanical Engineering Hang, Ta-Chun Tedford, Nathan C. Reddy, Raven J. Rimchala, Tharathorn White, Forest M. Kamm, Roger Dale Lauffenburger, Douglas A. The process of angiogenesis is under complex regulation in adult organisms, particularly as it often occurs in an inflammatory post-wound environment. As such, there are many impacting factors that will regulate the generation of new blood vessels which include not only pro-angiogenic growth factors such as vascular endothelial growth factor, but also angiostatic factors. During initial postwound hemostasis, a large initial bolus of platelet factor 4 is released into localized areas of damage before progression of wound healing toward tissue homeostasis. Because of its early presence and high concentration, the angiostatic chemokine platelet factor 4, which can induce endothelial anoikis, can strongly affect angiogenesis. In our work, we explored signaling crosstalk interactions between vascular endothelial growth factor and platelet factor 4 using phosphotyrosine-enriched mass spectrometry methods on human dermal microvascular endothelial cells cultured under conditions facilitating migratory sprouting into collagen gel matrices. We developed new methods to enable mass spectrometry-based phosphorylation analysis of primary cells cultured on collagen gels, and quantified signaling pathways over the first 48 h of treatment with vascular endothelial growth factor in the presence or absence of platelet factor 4. By observing early and late signaling dynamics in tandem with correlation network modeling, we found that platelet factor 4 has significant crosstalk with vascular endothelial growth factor by modulating cell migration and polarization pathways, centered around P38α MAPK, Src family kinases Fyn and Lyn, along with FAK. Interestingly, we found EphA2 correlational topology to strongly involve key migration-related signaling nodes after introduction of platelet factor 4, indicating an influence of the angiostatic factor on this ambiguous but generally angiogenic signal in this complex environment. National Science Foundation (U.S.) (NSF EFRI grant 735997) National Institutes of Health (U.S.) (NIH Cell Migration Consortium grant GM06346) National Institutes of Health (U.S.) (NIH Cell Decision Processes Center grant GM68762) National Institutes of Health (U.S.) (NIH grant GM69668) National Institutes of Health (U.S.) (NIH grant GM81336) 2014-09-03T14:52:21Z 2014-09-03T14:52:21Z 2013-09 2013-09 Article http://purl.org/eprint/type/JournalArticle 1535-9476 1535-9484 http://hdl.handle.net/1721.1/89149 Hang, T.-C., N. C. Tedford, R. J. Reddy, T. Rimchala, A. Wells, F. M. White, R. D. Kamm, and D. A. Lauffenburger. “Vascular Endothelial Growth Factor (VEGF) and Platelet (PF-4) Factor 4 Inputs Modulate Human Microvascular Endothelial Signaling in a Three-Dimensional Matrix Migration Context.” Molecular & Cellular Proteomics 12, no. 12 (December 1, 2013): 3704–3718. https://orcid.org/0000-0002-1545-1651 https://orcid.org/0000-0002-7232-304X https://orcid.org/0000-0002-3856-7454 en_US http://dx.doi.org/10.1074/mcp.M113.030528 Molecular & Cellular Proteomics Creative Commons Attribution http://creativecommons.org/licenses/by/3.0/ application/pdf American Society for Biochemistry and Molecular Biology (ASBMB) Molecular & Cellular Proteomics |
| spellingShingle | Hang, Ta-Chun Tedford, Nathan C. Reddy, Raven J. Rimchala, Tharathorn Wells, Alan White, Forest M. Kamm, Roger Dale Lauffenburger, Douglas A. Vascular Endothelial Growth Factor (VEGF) and Platelet (PF-4) Factor 4 Inputs Modulate Human Microvascular Endothelial Signaling in a Three-Dimensional Matrix Migration Context |
| title | Vascular Endothelial Growth Factor (VEGF) and Platelet (PF-4) Factor 4 Inputs Modulate Human Microvascular Endothelial Signaling in a Three-Dimensional Matrix Migration Context |
| title_full | Vascular Endothelial Growth Factor (VEGF) and Platelet (PF-4) Factor 4 Inputs Modulate Human Microvascular Endothelial Signaling in a Three-Dimensional Matrix Migration Context |
| title_fullStr | Vascular Endothelial Growth Factor (VEGF) and Platelet (PF-4) Factor 4 Inputs Modulate Human Microvascular Endothelial Signaling in a Three-Dimensional Matrix Migration Context |
| title_full_unstemmed | Vascular Endothelial Growth Factor (VEGF) and Platelet (PF-4) Factor 4 Inputs Modulate Human Microvascular Endothelial Signaling in a Three-Dimensional Matrix Migration Context |
| title_short | Vascular Endothelial Growth Factor (VEGF) and Platelet (PF-4) Factor 4 Inputs Modulate Human Microvascular Endothelial Signaling in a Three-Dimensional Matrix Migration Context |
| title_sort | vascular endothelial growth factor vegf and platelet pf 4 factor 4 inputs modulate human microvascular endothelial signaling in a three dimensional matrix migration context |
| url | http://hdl.handle.net/1721.1/89149 https://orcid.org/0000-0002-1545-1651 https://orcid.org/0000-0002-7232-304X https://orcid.org/0000-0002-3856-7454 |
| work_keys_str_mv | AT hangtachun vascularendothelialgrowthfactorvegfandplateletpf4factor4inputsmodulatehumanmicrovascularendothelialsignalinginathreedimensionalmatrixmigrationcontext AT tedfordnathanc vascularendothelialgrowthfactorvegfandplateletpf4factor4inputsmodulatehumanmicrovascularendothelialsignalinginathreedimensionalmatrixmigrationcontext AT reddyravenj vascularendothelialgrowthfactorvegfandplateletpf4factor4inputsmodulatehumanmicrovascularendothelialsignalinginathreedimensionalmatrixmigrationcontext AT rimchalatharathorn vascularendothelialgrowthfactorvegfandplateletpf4factor4inputsmodulatehumanmicrovascularendothelialsignalinginathreedimensionalmatrixmigrationcontext AT wellsalan vascularendothelialgrowthfactorvegfandplateletpf4factor4inputsmodulatehumanmicrovascularendothelialsignalinginathreedimensionalmatrixmigrationcontext AT whiteforestm vascularendothelialgrowthfactorvegfandplateletpf4factor4inputsmodulatehumanmicrovascularendothelialsignalinginathreedimensionalmatrixmigrationcontext AT kammrogerdale vascularendothelialgrowthfactorvegfandplateletpf4factor4inputsmodulatehumanmicrovascularendothelialsignalinginathreedimensionalmatrixmigrationcontext AT lauffenburgerdouglasa vascularendothelialgrowthfactorvegfandplateletpf4factor4inputsmodulatehumanmicrovascularendothelialsignalinginathreedimensionalmatrixmigrationcontext |