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A mechanosensitive transcriptional mechanism that controls angiogenesis

Abstract

Angiogenesis is controlled by physical interactions between cells and extracellular matrix as well as soluble angiogenic factors, such as VEGF. However, the mechanism by which mechanical signals integrate with other microenvironmental cues to regulate neovascularization remains unknown. Here we show that the Rho inhibitor, p190RhoGAP (also known as GRLF1), controls capillary network formation in vitro in human microvascular endothelial cells and retinal angiogenesis in vivo by modulating the balance of activities between two antagonistic transcription factors, TFII-I (also known as GTF2I) and GATA2, that govern gene expression of the VEGF receptor VEGFR2 (also known as KDR). Moreover, this new angiogenesis signalling pathway is sensitive to extracellular matrix elasticity as well as soluble VEGF. This is, to our knowledge, the first known functional cross-antagonism between transcription factors that controls tissue morphogenesis, and that responds to both mechanical and chemical cues.

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Figure 1: TFII-I and GATA2 control VEGFR2 expression via p190RhoGAP.
Figure 2: Matrix elasticity controls VEGFR2 expression via TFII-I and GATA2.
Figure 3: Antagonism between GATA2 and TFII-I controls capillary cell migration and tube formation in vitro.
Figure 4: Matrix elasticity controls vessel formation via TFII-I and GATA2 in vivo.
Figure 5: TFII-I, GATA2 and p190RhoGAP regulate retinal vessel formation in vivo.

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Zixuan Zhao, Xinyi Chen, … Hanry Yu

References

  1. Ferrara, N., Gerber, H. P. & LeCouter, J. The biology of VEGF and its receptors. Nature Med. 9, 669–676 (2003)

    Article  CAS  Google Scholar 

  2. Ferrara, N., Mass, R. D., Campa, C. & Kim, R. Targeting VEGF-A to treat cancer and age-related macular degeneration. Annu. Rev. Med. 58, 491–504 (2007)

    Article  CAS  Google Scholar 

  3. Ingber, D. E. & Folkman, J. Mechanochemical switching between growth and differentiation during fibroblast growth factor-stimulated angiogenesis in vitro: role of extracellular matrix. J. Cell Biol. 109, 317–330 (1989)

    Article  CAS  Google Scholar 

  4. Chen, C. S., Mrksich, M., Huang, S., Whitesides, G. M. & Ingber, D. E. Geometric control of cell life and death. Science 276, 1425–1428 (1997)

    Article  CAS  Google Scholar 

  5. Dike, L. E. et al. Geometric control of switching between growth, apoptosis, and differentiation during angiogenesis using micropatterned substrates. In Vitro Cell. Dev. Biol. Anim. 35, 441–448 (1999)

    Article  CAS  Google Scholar 

  6. Parker, K. K. et al. Directional control of lamellipodia extension by constraining cell shape and orienting cell tractional forces. FASEB J. 16, 1195–1204 (2002)

    Article  CAS  Google Scholar 

  7. Matthews, B. D., Overby, D. R., Mannix, R. & Ingber, D. E. Cellular adaptation to mechanical stress: role of integrins, Rho, cytoskeletal tension and mechanosensitive ion channels. J. Cell Sci. 119, 508–518 (2006)

    Article  CAS  Google Scholar 

  8. Kumar, S. et al. Viscoelastic retraction of single living stress fibers and its impact on cell shape, cytoskeletal organization, and extracellular matrix mechanics. Biophys. J. 90, 3762–3773 (2006)

    Article  CAS  Google Scholar 

  9. Moore, K. A. et al. Control of basement membrane remodeling and epithelial branching morphogenesis in embryonic lung by Rho and cytoskeletal tension. Dev. Dyn. 232, 268–281 (2005)

    Article  CAS  Google Scholar 

  10. Huang, S. & Ingber, D. E. The structural and mechanical complexity of cell-growth control. Nature Cell Biol. 1, E131–E138 (1999)

    Article  CAS  Google Scholar 

  11. Folkman, J. & Moscona, A. Role of cell shape in growth control. Nature 273, 345–349 (1978)

    Article  CAS  Google Scholar 

  12. Folkman, J. & Kalluri, R. Cancer without disease. Nature 427, 787 (2004)

    Article  CAS  Google Scholar 

  13. Matsumoto, T. & Claesson-Welsh, L. VEGF receptor signal transduction. Sci. STKE 2001, re21 (2001)

    CAS  Google Scholar 

  14. Wong, C. G., Rich, K. A., Liaw, L. H., Hsu, H. T. & Berns, M. W. Intravitreal VEGF and bFGF produce florid retinal neovascularization and hemorrhage in the rabbit. Curr. Eye Res. 22, 140–147 (2001)

    Article  CAS  Google Scholar 

  15. Mammoto, A., Huang, S., Moore, K., Oh, P. & Ingber, D. E. Role of RhoA, mDia, and ROCK in cell shape-dependent control of the Skp2-p27kip1 pathway and the G1/S transition. J. Biol. Chem. 279, 26323–26330 (2004)

    Article  CAS  Google Scholar 

  16. Mammoto, A., Huang, S. & Ingber, D. E. Filamin links cell shape and cytoskeletal structure to Rho regulation by controlling accumulation of p190RhoGAP in lipid rafts. J. Cell Sci. 120, 456–467 (2007)

    Article  CAS  Google Scholar 

  17. Jiang, W. et al. An FF domain-dependent protein interaction mediates a signaling pathway for growth factor-induced gene expression. Mol. Cell 17, 23–35 (2005)

    Article  CAS  Google Scholar 

  18. Jackson, T. A., Taylor, H. E., Sharma, D., Desiderio, S. & Danoff, S. K. Vascular endothelial growth factor receptor-2: counter-regulation by the transcription factors, TFII-I and TFII-IRD1. J. Biol. Chem. 280, 29856–29863 (2005)

    Article  CAS  Google Scholar 

  19. Roy, A. L. Biochemistry and biology of the inducible multifunctional transcription factor TFII-I. Gene 274, 1–13 (2001)

    Article  CAS  Google Scholar 

  20. Francke, U. Williams–Beuren syndrome: genes and mechanisms. Hum. Mol. Genet. 8, 1947–1954 (1999)

    Article  CAS  Google Scholar 

  21. Roy, A. L. Signal-induced functions of the transcription factor TFII-I. Biochim. Biophys. Acta 1769, 613–621 (2007)

    Article  CAS  Google Scholar 

  22. Patterson, C. et al. Cloning and functional analysis of the promoter for KDR/flk-1, a receptor for vascular endothelial growth factor. J. Biol. Chem. 270, 23111–23118 (1995)

    Article  CAS  Google Scholar 

  23. Minami, T., Rosenberg, R. D. & Aird, W. C. Transforming growth factor-β1-mediated inhibition of the flk-1/KDR gene is mediated by a 5′-untranslated region palindromic GATA site. J. Biol. Chem. 276, 5395–5402 (2001)

    Article  CAS  Google Scholar 

  24. Minami, T. et al. Interaction between hex and GATA transcription factors in vascular endothelial cells inhibits flk-1/KDR-mediated vascular endothelial growth factor signaling. J. Biol. Chem. 279, 20626–20635 (2004)

    Article  CAS  Google Scholar 

  25. Cantor, A. B. & Orkin, S. H. Hematopoietic development: a balancing act. Curr. Opin. Genet. Dev. 11, 513–519 (2001)

    Article  CAS  Google Scholar 

  26. Grogan, J. L. & Locksley, R. M. T helper cell differentiation: on again, off again. Curr. Opin. Immunol. 14, 366–372 (2002)

    Article  CAS  Google Scholar 

  27. Pai, S. Y., Truitt, M. L. & Ho, I. C. GATA-3 deficiency abrogates the development and maintenance of T helper type 2 cells. Proc. Natl Acad. Sci. USA 101, 1993–1998 (2004)

    Article  CAS  Google Scholar 

  28. Kouros-Mehr, H., Slorach, E. M., Sternlicht, M. D. & Werb, Z. GATA-3 maintains the differentiation of the luminal cell fate in the mammary gland. Cell 127, 1041–1055 (2006)

    Article  CAS  Google Scholar 

  29. Su, Z. J. et al. A vascular cell-restricted RhoGAP, p73RhoGAP, is a key regulator of angiogenesis. Proc. Natl Acad. Sci. USA 101, 12212–12217 (2004)

    Article  CAS  Google Scholar 

  30. Arthur, W. T., Petch, L. A. & Burridge, K. Integrin engagement suppresses RhoA activity via a c-Src-dependent mechanism. Curr. Biol. 10, 719–722 (2000)

    Article  CAS  Google Scholar 

  31. Robinson, C. J. & Stringer, S. E. The splice variants of vascular endothelial growth factor (VEGF) and their receptors. J. Cell Sci. 114, 853–865 (2001)

    CAS  Google Scholar 

  32. Sheibani, N. & Frazier, W. A. Down-regulation of platelet endothelial cell adhesion molecule-1 results in thrombospondin-1 expression and concerted regulation of endothelial cell phenotype. Mol. Biol. Cell 9, 701–713 (1998)

    Article  CAS  Google Scholar 

  33. Numaguchi, Y. et al. Caldesmon-dependent switching between capillary endothelial cell growth and apoptosis through modulation of cell shape and contractility. Angiogenesis 6, 55–64 (2003)

    Article  CAS  Google Scholar 

  34. Polte, T. R., Eichler, G. S., Wang, N. & Ingber, D. E. Extracellular matrix controls myosin light chain phosphorylation and cell contractility through modulation of cell shape and cytoskeletal prestress. Am. J. Physiol. Cell Physiol. 286, C518–C528 (2004)

    Article  CAS  Google Scholar 

  35. Pierce, E. A., Avery, R. L., Foley, E. D., Aiello, L. P. & Smith, L. E. Vascular endothelial growth factor/vascular permeability factor expression in a mouse model of retinal neovascularization. Proc. Natl Acad. Sci. USA 92, 905–909 (1995)

    Article  CAS  Google Scholar 

  36. Stalmans, I. et al. Arteriolar and venular patterning in retinas of mice selectively expressing VEGF isoforms. J. Clin. Invest. 109, 327–336 (2002)

    Article  CAS  Google Scholar 

  37. Mammoto, T. et al. Angiopoietin-1 requires p190RhoGAP to protect against vascular leakage in vivo . J. Biol. Chem. 282, 23910–23918 (2007)

    Article  CAS  Google Scholar 

  38. Singh, H., Medina, K. L. & Pongubala, J. M. Contingent gene regulatory networks and B cell fate specification. Proc. Natl Acad. Sci. USA 102, 4949–4953 (2005)

    Article  CAS  Google Scholar 

  39. Swiers, G., Patient, R. & Loose, M. Genetic regulatory networks programming hematopoietic stem cells and erythroid lineage specification. Dev. Biol. 294, 525–540 (2006)

    Article  CAS  Google Scholar 

  40. Gottgens, B. et al. Establishing the transcriptional programme for blood: the SCL stem cell enhancer is regulated by a multiprotein complex containing Ets and GATA factors. EMBO J. 21, 3039–3050 (2002)

    Article  CAS  Google Scholar 

  41. Engler, A. J., Sen, S., Sweeney, H. L. & Discher, D. E. Matrix elasticity directs stem cell lineage specification. Cell 126, 677–689 (2006)

    Article  CAS  Google Scholar 

  42. Clark, E. R. & Clark, E. L. Microscopic observations on the growth of blood capillaries in the living mammal. Am. J. Anat. 64, 251–301 (1939)

    Article  Google Scholar 

  43. Carmeliet, P. & Tessier-Lavigne, M. Common mechanisms of nerve and blood vessel wiring. Nature 436, 193–200 (2005)

    Article  CAS  Google Scholar 

  44. Pelham, R. J. & Wang, Y. Cell locomotion and focal adhesions are regulated by substrate flexibility. Proc. Natl Acad. Sci. USA 94, 13661–13665 (1997)

    Article  CAS  Google Scholar 

  45. Wang, N. et al. Cell prestress. I. Stiffness and prestress are closely associated in adherent contractile cells. Am. J. Physiol. Cell Physiol. 282, C606–C616 (2002)

    Article  CAS  Google Scholar 

  46. Yung, C. W. et al. Transglutaminase crosslinked gelatin as a tissue engineering scaffold. J. Biomed. Mater. Res. A 83, 1039–1046 (2007)

    Article  CAS  Google Scholar 

  47. Connor, K. M. et al. Increased dietary intake of Ω-3-polyunsaturated fatty acids reduces pathological retinal angiogenesis. Nature Med. 13, 868–873 (2007)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank T. Polte, E. Pravda, M. de Bruijn and K. Johnson for their technical suggestions and assistance, T. Nakano and H. Sabe for providing plasmids, the National Institutes of Health (NIH) for providing VEGF, and D. Weitz for providing assistance with rheometry measurements. This work was supported by funds from the NIH (to D.E.I., L.E.H.S. and K.M.C.), V. Kann Rasmussen Foundation (to L.E.H.S.), Children’s Hospital Mental Retardation and Developmental Disabilities Research Center (to L.E.H.S.), a Research to Prevent Blindness Lew Wasserman Merit Award (to L.E.H.S.), American Heart Association (to A.M.), and a Children’s Hospital House Officer Development Award (to A.M.); D.E.I. is a recipient of a DoD Breast Cancer Innovator Award.

Author Contributions A.M. conceived the experiments, performed experiments, designed research and analysed data with assistance from K.M.C., T.M., C.W.Y., D.H., C.M.A., G.M., L.E.H.S. and D.E.I. A.M. wrote the manuscript with D.E.I., with input from L.E.H.S.

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Correspondence to Donald E. Ingber.

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Mammoto, A., Connor, K., Mammoto, T. et al. A mechanosensitive transcriptional mechanism that controls angiogenesis. Nature 457, 1103–1108 (2009). https://doi.org/10.1038/nature07765

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