Loss of the tumor suppressor Vhlh leads to upregulation of Cxcr4 and rapidly progressive glomerulonephritis in mice

Abstract

Rapidly progressive glomerulonephritis (RPGN) is a clinical syndrome characterized by loss of renal function within days to weeks and by glomerular crescents on biopsy. The pathogenesis of this disease is unclear, but circulating factors are believed to have a major role1,2. Here, we show that deletion of the Von Hippel–Lindau gene (Vhlh) from intrinsic glomerular cells of mice is sufficient to initiate a necrotizing crescentic glomerulonephritis and the clinical features that accompany RPGN. Loss of Vhlh leads to stabilization of hypoxia-inducible factor α subunits (HIFs). Using gene expression profiling, we identified de novo expression of the HIF target gene Cxcr4 (ref. 3) in glomeruli from both mice and humans with RPGN. The course of RPGN is markedly improved in mice treated with a blocking antibody to Cxcr4, whereas overexpression of Cxcr4 alone in podocytes of transgenic mice is sufficient to cause glomerular disease. Collectively, these results indicate an alternative mechanism for the pathogenesis of RPGN and glomerular disease in an animal model and suggest novel molecular pathways for intervention in this disease.

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Figure 1: Selective deletion of Vhlh from podocytes leads to RPGN.
Figure 2: Podocyte proliferation initiates crescent formation.
Figure 3: Increased Cxcr4 is functionally important for the RPGN phenotype.
Figure 4: De novo expression of CXCR4 in mice and humans with glomerular disease.

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Acknowledgements

The authors thank D. Vukasovic for secretarial assistance and the Centre for Modelling Human Diseases for biochemical assays in the mice. We also thank B. Pressler (University of North Carolina at Chapel Hill) for performing ANCA assays, Y. Wang (Samuel Lunenfeld Research Institute) for help in LCM isolation, S. Peiper (Institute of Molecular Medicine and Genetics, Georgia) for providing the constitutively active Cxcr4 constructs, V. Eremina for technical assistance, K. Kamel (St. Michael's Hospital, Toronto) and D. Cattran (Toronto Hospital) for critically reviewing the manuscript. S.E.Q. is the recipient of a Canada Research Chair Tier II, and a Premier's Research of Excellence Award. This work was funded by Canadian Institute of Health Research grant MOP 77756, National Cancer Institute of Canada grant #16002 and Emerald Foundation grant (to S.E.Q.).

Author information

Correspondence to Susan E Quaggin.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

The renal glomerulus. (PDF 68 kb)

Supplementary Fig. 2

Generation of transgenic mouse lines. (PDF 340 kb)

Supplementary Fig. 3

PCNA staining. (PDF 94 kb)

Supplementary Fig. 4

Hif1a protein is increased in podocytes from VhlhloxP/loxP Pod-Cre mice. (PDF 165 kb)

Supplementary Fig. 5

Glomerular expression of Hif target genes. (PDF 259 kb)

Supplementary Fig. 6

Expression of the Hif target gene Sdf1 in glomeruli of wild-type and mutant mice. (PDF 119 kb)

Supplementary Fig. 7

Kaplan-Meier survival curves for mutant Vhlh mice treated with blocking antibody to Cxcr4 or vehicle. (PDF 48 kb)

Supplementary Fig. 8

mRNA expression analysis of VHL-HIF pathway molecular targets in glomeruli from patients with pauci-immune RPGN, IgA nephritis or no glomerular disease. (PDF 159 kb)

Supplementary Table 1

Gene expression patterns in human RPGN compared with expression in glomeruli from VhlhloxP/loxP Pod-Cre mice. (PDF 96 kb)

Supplementary Note (PDF 29 kb)

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