Glomerular-specific alterations of VEGF-A expression lead to distinct congenital and acquired renal diseases

Vera Eremina

Manish Sood

Jody Haigh

András Nagy

^{The Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada}^{Princess Margaret Hospital, University Health Network, Department of Pathology, Toronto, Ontario, Canada}^{Department of Molecular Oncology, Genentech Inc., South San Francisco, California, USA}^{Department of Medicine, Washington University School of Medicine, Renal Division, St. Louis, Missouri, USA}^{St. Michael’s Hospital, Toronto, Ontario, Canada}

Address correspondence to: Susan E. Quaggin, The Samuel Lunenfeld Research Institute, Room 871Q, Mount Sinai Hospital, 600 University Avenue, Toronto, Ontario M5G 1X5, Canada. Phone: (416) 586-4800 ext. 2859; Fax: (416) 586-8588; E-mail: ac.no.irhsm@niggauq.

Received 2002 Nov 18; Accepted 2003 Jan 15.

Abstract

Kidney disease affects over 20 million people in the United States alone. Although the causes of renal failure are diverse, the glomerular filtration barrier is often the target of injury. Dysregulation of VEGF expression within the glomerulus has been demonstrated in a wide range of primary and acquired renal diseases, although the significance of these changes is unknown. In the glomerulus, VEGF-A is highly expressed in podocytes that make up a major portion of the barrier between the blood and urinary spaces. In this paper, we show that glomerular-selective deletion or overexpression of VEGF-A leads to glomerular disease in mice. Podocyte-specific heterozygosity for VEGF-A resulted in renal disease by 2.5 weeks of age, characterized by proteinuria and endotheliosis, the renal lesion seen in preeclampsia. Homozygous deletion of VEGF-A in glomeruli resulted in perinatal lethality. Mutant kidneys failed to develop a filtration barrier due to defects in endothelial cell migration, differentiation, and survival. In contrast, podocyte-specific overexpression of the VEGF-164 isoform led to a striking collapsing glomerulopathy, the lesion seen in HIV-associated nephropathy. Our data demonstrate that tight regulation of VEGF-A signaling is critical for establishment and maintenance of the glomerular filtration barrier and strongly supports a pivotal role for VEGF-A in renal disease.

Abstract

Acknowledgments

We gratefully acknowledge the technical support of Lois Schwartz for generation of transgenic mice, Doug Holmyard for EM processing and images, the Toronto Centre for Comparative Models of Human Disease for help with biochemical analysis of the mice, and Dragana Vukasovic for excellent secretarial assistance. We also thank Janet Rossant and Jordan Kreidberg for critically reviewing the manuscript and Wilhelm Kriz for invaluable assistance. S.E. Quaggin is the recipient of a Canada Research Chair. This work was funded by NIH grant 5 R21 DK-59148-02 and a Kidney Foundation of Canada Grant (to S.E. Quaggin). This work was inspired by the courage of Kelly Kalt (1983–2000).

Acknowledgments

Footnotes

See the related Commentary beginning on page 600.

Conflict of interest: The authors have declared that no conflict of interest exists.

Nonstandard abbreviations used: glomerular basement membrane (GBM); fetal liver kinase 1 (Flk1); fms-like tyrosine kinase 1 (Flt1); HIV-associated nephropathy (HIVAN); Wilms tumor suppressor gene (WT1); α-smooth muscle actin (VSMA).

Footnotes

References

1. Mundel P, Reiser JNew aspects of podocyte cell biology. Kidney Blood Press. Res.1997;20:173–176.[PubMed]
2. Abrahamson DRGlomerulogenesis in the developing kidney. Semin. Nephrol.1991;11:375–389.[PubMed]
3. Kriz W, Lemley KThe role of the podocyte in glomerulosclerosis. Curr. Opin. Nephrol. Hypertens.1999;8:489–497.[PubMed]
4. Kriz WEvolving role of the podocyte in chronic renal failure. Kidney Blood Press. Res.1997;20:180–183.[PubMed]
5. Shirato I, et al The development of focal segmental glomerulosclerosis in masugi nephritis is based on progressive podocyte damage. Virchows Arch.1996;429:255–273.[PubMed]
6. Coimbra TM, et al Early events leading to renal injury in obese Zucker (fatty) rats with type II diabetes. Kidney Int.2000;57:167–182.[PubMed]
7. Floege J, et al Age-related glomerulosclerosis and interstitial fibrosis in Milan normotensive rats: a podocyte disease. Kidney Int.1997;51:230–243.[PubMed]
8. Pagtalunan ME, et al Podocyte loss and progressive glomerular injury in type II diabetes. J. Clin. Invest.1997;99:342–348.
9. Robert B, Zhao X, Abrahamson DRCoexpression of neuropilin-1, flk1, and VEGF(164) in developing and mature mouse kidney glomeruli. Am. J. Physiol. Renal Physiol.2000;279:F275–F282.[PubMed]
10. Saxen L, Sariola HOrganogenesis of the kidney. Pediatr. Nephrol.1987;1:385–392.[PubMed]
11. Carmeliet P, et al Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature.1996;380:435–439.[PubMed]
12. Ferrara N, et al Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene. Nature.1996;380:439–442.[PubMed]
13. Somlo S, Mundel PGetting a foothold in nephrotic syndrome. Nat. Genet.2000;24:333–335.[PubMed]
14. Laurinavicius A, Hurwitz S, Rennke HGCollapsing glomerulopathy in HIV and non-HIV patients: a clinicopathological and follow-up study. Kidney Int.1999;56:2203–2213.[PubMed]
15. Novak A, Guo C, Yang W, Nagy A, Lobe CGZ/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon Cre-mediated excision. Genesis.2000;28:147–155.[PubMed]
16. Gerber HP, et al VEGF is required for growth and survival in neonatal mice. Development.1999;126:1149–1159.[PubMed]
17. Eremina V, Wong MA, Cui S, Schwartz L, Quaggin SEGlomerular-specific gene excision in vivo. J. Am. Soc. Nephrol.2002;13:788–793.[PubMed]
18. Laemmli UKCleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature.1970;227:680–685.[PubMed]
19. Wong MA, Cui S, Quaggin SEIdentification and characterization of a glomerular-specific promoter from the human nephrin gene. Am. J. Physiol. Renal Physiol.2000;279:F1027–F1032.[PubMed]
20. Miner JH, Li CDefective glomerulogenesis in the absence of laminin alpha5 demonstrates a developmental role for the kidney glomerular basement membrane. Dev. Biol.2000;217:278–289.[PubMed]
21. Kincaid-Smith PThe renal lesion of preeclampsia revisited. Am. J. Kidney Dis.1991;17:144–148.[PubMed]
22. McDonald R, Wiggelinkhuizen J, Kaschula ROThe nephrotic syndrome in very young infants. Am. J. Dis. Child.1971;122:507–512.[PubMed]
23. Esser S, et al Vascular endothelial growth factor induces endothelial fenestrations in vitro. J. Cell Biol.1998;140:947–959.
24. Roberts WG, Palade GEIncreased microvascular permeability and endothelial fenestration induced by vascular endothelial growth factor. J. Cell Sci.1995;108:2369–2379.[PubMed]
25. Bielecki DA, Klonowska-Dziatkiewicz E, Jarocki S, Urban JGrowth factors in pregnancy complications with preeclampsia. Ginekol Pol.2002;73:422–429.[PubMed]
26. Bachelder RE, et al Vascular endothelial growth factor is an autocrine survival factor for neuropilin-expressing breast carcinoma cells. Cancer Res.2001;61:5736–5740.[PubMed]
27. Kitamoto Y, Tokunaga H, Tomita KVascular endothelial growth factor is an essential molecule for mouse kidney development: glomerulogenesis and nephrogenesis. J. Clin. Invest.1997;99:2351–2357.
28. Mattot V, et al Loss of the VEGF(164) and VEGF(188) isoforms impairs postnatal glomerular angiogenesis and renal arteriogenesis in mice. J. Am. Soc. Nephrol.2002;13:1548–1560.[PubMed]
29. Miquerol L, Gertsenstein M, Harpal K, Rossant J, Nagy AMultiple developmental roles of VEGF suggested by a LacZ-tagged allele. Dev. Biol.1999;212:307–322.[PubMed]
30. Gengrinovitch S, et al Glypican-1 is a VEGF165 binding proteoglycan that acts as an extracellular chaperone for VEGF165. J. Biol. Chem.1999;274:10816–10822.[PubMed]
31. Drake CJ, Little CDExogenous vascular endothelial growth factor induces malformed and hyperfused vessels during embryonic neovascularization. Proc. Natl. Acad. Sci. U. S. A.1995;92:7657–7661.
32. Drake CJ, Little CDVEGF and vascular fusion: implications for normal and pathological vessels. J. Histochem. Cytochem.1999;47:1351–1356.[PubMed]
33. Morini M, et al Kaposi’s sarcoma cells of different etiologic origins respond to HIV-Tat through the Flk-1/KDR (VEGFR-2): relevance in AIDS-KS pathology. Biochem. Biophys. Res. Commun.2000;273:267–271.[PubMed]
34. Ganju RK, et al Human immunodeficiency virus tat modulates the Flk-1/KDR receptor, mitogen-activated protein kinases, and components of focal adhesion in Kaposi’s sarcoma cells. J. Virol.1998;72:6131–6137.
35. Vene R, Benelli R, Noonan DM, Albini AHIV-Tat dependent chemotaxis and invasion, key aspects of tat mediated pathogenesis. Clin. Exp. Metastasis.2000;18:533–538.[PubMed]
36. Marras D, et al Replication and compartmentalization of HIV-1 in kidney epithelium of patients with HIV-associated nephropathy. Nat. Med.2002;8:522–526.[PubMed]
37. Barisoni L, Kriz W, Mundel P, D’Agati VThe dysregulated podocyte phenotype: a novel concept in the pathogenesis of collapsing idiopathic focal segmental glomerulosclerosis and HIV-associated nephropathy. J. Am. Soc. Nephrol.1999;10:51–61.[PubMed]
38. Bruggeman LA, et al Nephropathy in human immunodeficiency virus-1 transgenic mice is due to renal transgene expression. J. Clin. Invest.1997;100:84–92.
39. Conaldi PG, et al Human immunodeficiency virus-1 tat induces hyperproliferation and dysregulation of renal glomerular epithelial cells. Am. J. Pathol.2002;161:53–61.
40. Husain M, et al HIV-1 Nef induces proliferation and anchorage-independent growth in podocytes. J. Am. Soc. Nephrol.2002;13:1806–1815.[PubMed]
41. Shahbazi M, et al Vascular endothelial growth factor gene polymorphisms are associated with acute renal allograft rejection. J. Am. Soc. Nephrol.2002;13:260–264.[PubMed]
42. Kim YG, et al Vascular endothelial growth factor accelerates renal recovery in experimental thrombotic microangiopathy. Kidney Int.2000;58:2390–2399.[PubMed]
43. Cha DR, et al Role of vascular endothelial growth factor in diabetic nephropathy. Kidney Int. Suppl.2000;77:S104–S112.[PubMed]
44. Ostendorf T, et al VEGF(165) mediates glomerular endothelial repair. J. Clin. Invest.1999;104:913–923.