A systems approach identifies HIPK2 as a key regulator of kidney fibrosis

Abstract

Kidney fibrosis is a common process that leads to the progression of various types of kidney disease. We used an integrated computational and experimental systems biology approach to identify protein kinases that regulate gene expression changes in the kidneys of human immunodeficiency virus (HIV) transgenic mice (Tg26 mice), which have both tubulointerstitial fibrosis and glomerulosclerosis. We identified homeo-domain interacting protein kinase 2 (HIPK2) as a key regulator of kidney fibrosis. HIPK2 was upregulated in the kidneys of Tg26 mice and in those of patients with various kidney diseases. HIV infection increased the protein concentrations of HIPK2 by promoting oxidative stress, which inhibited the seven in absentia homolog 1 (SIAH1)-mediated proteasomal degradation of HIPK2. HIPK2 induced apoptosis and the expression of epithelial-to-mesenchymal transition markers in kidney epithelial cells by activating the p53, transforming growth factor β (TGF-β)–SMAD family member 3 (Smad3) and Wnt-Notch pathways. Knockout of HIPK2 improved renal function and attenuated proteinuria and kidney fibrosis in Tg26 mice, as well as in other murine models of kidney fibrosis. We therefore conclude that HIPK2 is a potential target for anti-fibrosis therapy.

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Figure 1: Identification of key signaling pathways activated in HIVAN.
Figure 2: HIV induces HIPK2 expression in kidney cells.
Figure 3: SIAH1 is an upstream regulator of HIPK2 expression.
Figure 4: HIPK2 mediates HIV-induced apoptosis and EMT marker expression in hRTECs.
Figure 5: HIPK2 mediates downstream signaling pathways in RTECs.
Figure 6: Knockout of HIPK2 prevents kidney injury in Tg26.

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Accessions

GenBank/EMBL/DDBJ

Gene Expression Omnibus

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Acknowledgements

The authors acknowledge and thank G.L. Gusella (Mount Sinai School of Medicine, New York) for providing the lentiviral construct, E. Huang (University of California San Francisco) for providing the HIPK2 KO mice, P. Mundel (Massachusetts General Hospital, Boston) for synaptopodin antibody and conditionally immortalized murine podocyte cell line, L. Holzman (University of Pennsylvania, Philadelphia) for nephrin antibody and R.H. Goodman (Oregon Health and Science University, Portland) for WT-HIPK2 and KD-HIPK2 constructs. We also thank R. Iyengar and P. Klotman for critical suggestions on experimental design and for other valuable contributions. J.C.H. is supported by US National Institutes of Health (NIH) grant 1R01DK078897 and a Veterans Affairs Merit Award; J.C.H. and A.M. are supported by NIH 1R01DK088541, NIH P01-DK-56492 and NIH 1RC4DK090860; A.M. is supported by NIH RC2OD006536 and 5P50GM071558; P.Y.C. is supported by NIH 5K08DK082760. N.C. is supported by Chinese 973 fund 2012CB517601.

Author information

J.C.H., A.M., P.Y.C., Y.J., N.C. and K.R. designed the research project; Y.J., K.R., Y.F., Y.Z., M.J.R., H.X. and P.Y.C. performed the experiments; P.Y.C. and Y.D. prepared lentiviral constructs for HIPK2; V.D. analyzed the pathologic findings; A.M., E.Y.C. and A.R.M. performed bioinformatics and systems biology analyses; J.C.H., P.Y.C. and A.M. wrote the manuscript, and all authors contributed to the preparation of the manuscript.

Correspondence to Avi Ma'ayan or John Cijiang He.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6, Supplementary Tables 1–6 and Supplementary Methods (PDF 4095 kb)

Supplementary Data

Summary of ChEA, TRANSFAC, Genes2Network and KEA for identification of key signaling pathways activated in Tg26 kidney (XLSX 134 kb)

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Jin, Y., Ratnam, K., Chuang, P. et al. A systems approach identifies HIPK2 as a key regulator of kidney fibrosis. Nat Med 18, 580–588 (2012). https://doi.org/10.1038/nm.2685

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