Cellular and Molecular Life Sciences

, Volume 69, Issue 13, pp 2217–2231 | Cite as

Airway basal cell vascular endothelial growth factor-mediated cross-talk regulates endothelial cell-dependent growth support of human airway basal cells

  • Giacomo Curradi
  • Matthew S. Walters
  • Bi-Sen Ding
  • Shahin Rafii
  • Neil R. Hackett
  • Ronald G. CrystalEmail author
Research Article


The human airway epithelium is a pseudostratified heterogenous layer comprised of ciliated, secretory, intermediate, and basal cells. As the stem/progenitor population of the airway epithelium, airway basal cells differentiate into ciliated and secretory cells to replenish the airway epithelium during physiological turnover and repair. Transcriptome analysis of airway basal cells revealed high expression of vascular endothelial growth factor A (VEGFA), a gene not typically associated with the function of this cell type. Using cultures of primary human airway basal cells, we demonstrate that basal cells express all of the three major isoforms of VEGFA (121, 165 and 189) but lack functional expression of the classical VEGFA receptors VEGFR1 and VEGFR2. The VEGFA is actively secreted by basal cells and while it appears to have no direct autocrine function on basal cell growth and proliferation, it functions in a paracrine manner to activate MAPK signaling cascades in endothelium via VEGFR2-dependent signaling pathways. Using a cytokine- and serum-free co-culture system of primary human airway basal cells and human endothelial cells revealed that basal cell-secreted VEGFA activated endothelium to express mediators that, in turn, stimulate and support basal cell proliferation and growth. These data demonstrate novel VEGFA-mediated cross-talk between airway basal cells and endothelium, the purpose of which is to modulate endothelial activation and in turn stimulate and sustain basal cell growth.


Basal cell Endothelium VEGF-A Airway Cross-talk Paracrine 



We thank R. Zwick for technical assistance, M. Staudt and J. Fuller for coordinating sample collection and N. Mohamed, D.N. McCarthy and R. Hamid for help in preparing this manuscript. These studies were supported, in part, by P50 HL084936, 1R01HL107882, UL1-RR024996 and UL1-RR024143.


  1. 1.
    Breeze RG, Wheeldon EB (1977) The cells of the pulmonary airways. Am Rev Respir Dis 116:705–777PubMedGoogle Scholar
  2. 2.
    Mercer RR, Russell ML, Roggli VL, Crapo JD (1994) Cell number and distribution in human and rat airways. Am J Respir Cell Mol Biol 10:613–624PubMedGoogle Scholar
  3. 3.
    Knight DA, Holgate ST (2003) The airway epithelium: structural and functional properties in health and disease. Respirology 8:432–446CrossRefPubMedGoogle Scholar
  4. 4.
    Thompson AB, Robbins RA, Romberger DJ, Sisson JH, Spurzem JR, Teschler H et al (1995) Immunological functions of the pulmonary epithelium. Eur Respir J 8:127–149CrossRefPubMedGoogle Scholar
  5. 5.
    Crystal RG, Randell SH, Engelhardt JF, Voynow J, Sunday ME (2008) Airway epithelial cells: current concepts and challenges. Proc Am Thorac Soc 5:772–777CrossRefPubMedGoogle Scholar
  6. 6.
    Evans MJ, Van Winkle LS, Fanucchi MV, Plopper CG (2001) Cellular and molecular characteristics of basal cells in airway epithelium. Exp Lung Res 27:401–415CrossRefPubMedGoogle Scholar
  7. 7.
    Hajj R, Baranek T, Le NR, Lesimple P, Puchelle E, Coraux C (2007) Basal cells of the human adult airway surface epithelium retain transit-amplifying cell properties. Stem Cells 25:139–148CrossRefPubMedGoogle Scholar
  8. 8.
    Rock JR, Onaitis MW, Rawlins EL, Lu Y, Clark CP, Xue Y et al (2009) Basal cells as stem cells of the mouse trachea and human airway epithelium. Proc Natl Acad Sci USA 106:12771–12775CrossRefPubMedGoogle Scholar
  9. 9.
    Rock JR, Randell SH, Hogan BL (2010) Airway basal stem cells: a perspective on their roles in epithelial homeostasis and remodeling. Dis Model Mech 3:545–556CrossRefPubMedGoogle Scholar
  10. 10.
    Rock JR, Gao X, Xue Y, Randell SH, Kong YY, Hogan BL (2011) Notch-dependent differentiation of adult airway basal stem cells. Cell Stem Cell 8:639–648CrossRefPubMedGoogle Scholar
  11. 11.
    Hackett NR, Shaykhiev R, Walters MS, Wang R, Zwick RK, Ferris B et al (2011) The human airway epithelial basal cell transcriptome. PLoS One 6:e18378CrossRefPubMedGoogle Scholar
  12. 12.
    Ferrara N, Gerber HP, LeCouter J (2003) The biology of VEGF and its receptors. Nat Med 9:669–676CrossRefPubMedGoogle Scholar
  13. 13.
    Grunewald FS, Prota AE, Giese A, Ballmer-Hofer K (2010) Structure–function analysis of VEGF receptor activation and the role of coreceptors in angiogenic signaling. Biochim Biophys Acta 1804:567–580PubMedGoogle Scholar
  14. 14.
    Tugues S, Koch S, Gualandi L, Li X, Claesson-Welsh L (2011) Vascular endothelial growth factors and receptors: anti-angiogenic therapy in the treatment of cancer. Mol Aspects Med 32:88–111CrossRefPubMedGoogle Scholar
  15. 15.
    Mura M, dos Santos CC, Stewart D, Liu M (2004) Vascular endothelial growth factor and related molecules in acute lung injury. J Appl Physiol 97:1605–1617CrossRefPubMedGoogle Scholar
  16. 16.
    Roy H, Bhardwaj S, Yla-Herttuala S (2006) Biology of vascular endothelial growth factors. FEBS Lett 580:2879–2887CrossRefPubMedGoogle Scholar
  17. 17.
    Woolard J, Bevan HS, Harper SJ, Bates DO (2009) Molecular diversity of VEGF-A as a regulator of its biological activity. Microcirculation 16:572–592CrossRefPubMedGoogle Scholar
  18. 18.
    Akeson A, Herman A, Wiginton D, Greenberg J (2010) Endothelial cell activation in a VEGF-A gradient: relevance to cell fate decisions. Microvasc Res 80:65–74CrossRefPubMedGoogle Scholar
  19. 19.
    Hackett NR, Heguy A, Harvey BG, O’Connor TP, Luettich K, Flieder DB et al (2003) Variability of antioxidant-related gene expression in the airway epithelium of cigarette smokers. Am J Respir Cell Mol Biol 29:331–343CrossRefPubMedGoogle Scholar
  20. 20.
    Harvey BG, Heguy A, Leopold PL, Carolan BJ, Ferris B, Crystal RG (2007) Modification of gene expression of the small airway epithelium in response to cigarette smoking. J Mol Med (Berl) 85:39–53CrossRefGoogle Scholar
  21. 21.
    Heguy A, Harvey BG, Leopold PL, Dolgalev I, Raman T, Crystal RG (2007) Responses of the human airway epithelium transcriptome to in vivo injury. Physiol Genomics 29:139–148PubMedGoogle Scholar
  22. 22.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  23. 23.
    Kobayashi H, Butler JM, O’Donnell R, Kobayashi M, Ding BS, Bonner B et al (2010) Angiocrine factors from Akt-activated endothelial cells balance self-renewal and differentiation of haematopoietic stem cells. Nat Cell Biol 12:1046–1056CrossRefPubMedGoogle Scholar
  24. 24.
    Soker S, Takashima S, Miao HQ, Neufeld G, Klagsbrun M (1998) Neuropilin-1 is expressed by endothelial and tumor cells as an isoform-specific receptor for vascular endothelial growth factor. Cell 92:735–745CrossRefPubMedGoogle Scholar
  25. 25.
    Suzuki M, Betsuyaku T, Nagai K, Fuke S, Nasuhara Y, Kaga K et al (2008) Decreased airway expression of vascular endothelial growth factor in cigarette smoke-induced emphysema in mice and COPD patients. Inhal Toxicol 20:349–359CrossRefPubMedGoogle Scholar
  26. 26.
    Seandel M, Butler JM, Kobayashi H, Hooper AT, White IA, Zhang F et al (2008) Generation of a functional and durable vascular niche by the adenoviral E4ORF1 gene. Proc Natl Acad Sci USA 105:19288–19293CrossRefPubMedGoogle Scholar
  27. 27.
    Kaner RJ, Crystal RG (2001) Compartmentalization of vascular endothelial growth factor to the epithelial surface of the human lung. Mol Med 7:240–246PubMedGoogle Scholar
  28. 28.
    Kranenburg AR, de Boer WI, Alagappan VK, Sterk PJ, Sharma HS (2005) Enhanced bronchial expression of vascular endothelial growth factor and receptors (Flk-1 and Flt-1) in patients with chronic obstructive pulmonary disease. Thorax 60:106–113CrossRefPubMedGoogle Scholar
  29. 29.
    Boussat S, Eddahibi S, Coste A, Fataccioli V, Gouge M, Housset B et al (2000) Expression and regulation of vascular endothelial growth factor in human pulmonary epithelial cells. Am J Physiol Lung Cell Mol Physiol 279:L371–L378PubMedGoogle Scholar
  30. 30.
    Medford AR, Douglas SK, Godinho SI, Uppington KM, Armstrong L, Gillespie KM et al (2009) Vascular endothelial growth factor (VEGF) isoform expression and activity in human and murine lung injury. Respir Res 10:27CrossRefPubMedGoogle Scholar
  31. 31.
    Alagappan VK, Willems-Widyastuti A, Seynhaeve AL, Garrelds IM, ten Hagen TL, Saxena PR et al (2007) Vasoactive peptides upregulate mRNA expression and secretion of vascular endothelial growth factor in human airway smooth muscle cells. Cell Biochem Biophys 47:109–118PubMedGoogle Scholar
  32. 32.
    Kamio K, Sato T, Liu X, Sugiura H, Togo S, Kobayashi T et al (2008) Prostacyclin analogs stimulate VEGF production from human lung fibroblasts in culture. Am J Physiol Lung Cell Mol Physiol 294:L1226–L1232CrossRefPubMedGoogle Scholar
  33. 33.
    Voelkel NF, Vandivier RW, Tuder RM (2006) Vascular endothelial growth factor in the lung. Am J Physiol Lung Cell Mol Physiol 290:L209–L221CrossRefPubMedGoogle Scholar
  34. 34.
    Ding BS, Nolan DJ, Guo P, Babazadeh AO, Cao Z, Rosenwaks Z et al (2011) Endothelial-derived angiocrine signals induce and sustain regenerative lung alveolarization. Cell 147:539–553CrossRefPubMedGoogle Scholar
  35. 35.
    Del Moral PM, Sala FG, Tefft D, Shi W, Keshet E, Bellusci S et al (2006) VEGF-A signaling through Flk-1 is a critical facilitator of early embryonic lung epithelial to endothelial crosstalk and branching morphogenesis. Dev Biol 290:177–188CrossRefPubMedGoogle Scholar
  36. 36.
    Yamamoto H, Yun EJ, Gerber HP, Ferrara N, Whitsett JA, Vu TH (2007) Epithelial–vascular cross-talk mediated by VEGF-A and HGF signaling directs primary septae formation during distal lung morphogenesis. Dev Biol 308:44–53CrossRefPubMedGoogle Scholar
  37. 37.
    Franzdottir SR, Axelsson IT, Arason AJ, Baldursson O, Gudjonsson T, Magnusson MK (2010) Airway branching morphogenesis in three dimensional culture. Respir Res 11:162CrossRefPubMedGoogle Scholar
  38. 38.
    Red-Horse K, Crawford Y, Shojaei F, Ferrara N (2007) Endothelium-microenvironment interactions in the developing embryo and in the adult. Dev Cell 12:181–194CrossRefPubMedGoogle Scholar
  39. 39.
    Butler JM, Nolan DJ, Vertes EL, Varnum-Finney B, Kobayashi H, Hooper AT et al (2010) Endothelial cells are essential for the self-renewal and repopulation of Notch-dependent hematopoietic stem cells. Cell Stem Cell 6:251–264CrossRefPubMedGoogle Scholar
  40. 40.
    Shen Q, Goderie SK, Jin L, Karanth N, Sun Y, Abramova N et al (2004) Endothelial cells stimulate self-renewal and expand neurogenesis of neural stem cells. Science 304:1338–1340CrossRefPubMedGoogle Scholar
  41. 41.
    Shopland DR (1995) Tobacco use and its contribution to early cancer mortality with a special emphasis on cigarette smoking. Environ Health Perspect 103(Suppl 8):131–142CrossRefPubMedGoogle Scholar
  42. 42.
    Hylkema MN, Sterk PJ, de Boer WI, Postma DS (2007) Tobacco use in relation to COPD and asthma. Eur Respir J 29:438–445CrossRefPubMedGoogle Scholar
  43. 43.
    Barnes PJ (2007) Chronic obstructive pulmonary disease: a growing but neglected global epidemic. PLoS Med 4:e112CrossRefPubMedGoogle Scholar
  44. 44.
    Auerbach O, Gere JB, Forman JB, Petrick TG, Smolin HJ, Muehsam GE et al (1957) Changes in the bronchial epithelium in relation to smoking and cancer of the lung a report of progress. N Engl J Med 256:97–104CrossRefPubMedGoogle Scholar
  45. 45.
    Merrick DT, Haney J, Petrunich S, Sugita M, Miller YE, Keith RL et al (2005) Overexpression of vascular endothelial growth factor and its receptors in bronchial dysplasia demonstrated by quantitative RT-PCR analysis. Lung Cancer 48:31–45CrossRefPubMedGoogle Scholar
  46. 46.
    Zanini A, Chetta A, Imperatori AS, Spanevello A, Olivieri D (2010) The role of the bronchial microvasculature in the airway remodelling in asthma and COPD. Respir Res 11:132CrossRefPubMedGoogle Scholar
  47. 47.
    Calabrese C, Bocchino V, Vatrella A, Marzo C, Guarino C, Mascitti S et al (2006) Evidence of angiogenesis in bronchial biopsies of smokers with and without airway obstruction. Respir Med 100:1415–1422CrossRefPubMedGoogle Scholar
  48. 48.
    Soltani A, Reid DW, Sohal SS, Wood-Baker R, Weston S, Muller HK et al (2010) Basement membrane and vascular remodelling in smokers and chronic obstructive pulmonary disease: a cross-sectional study. Respir Res 11:105CrossRefPubMedGoogle Scholar

Copyright information

© Springer Basel AG 2012

Authors and Affiliations

  • Giacomo Curradi
    • 1
    • 2
  • Matthew S. Walters
    • 1
  • Bi-Sen Ding
    • 1
  • Shahin Rafii
    • 1
  • Neil R. Hackett
    • 1
  • Ronald G. Crystal
    • 1
    Email author
  1. 1.Department of Genetic MedicineWeill Cornell Medical CollegeNew YorkUSA
  2. 2.Pneumo-Cardio-Thoracic Sciences Alma Mater StudiorumUniversity of BolognaBolognaItaly

Personalised recommendations