Nephronophthisis is an autosomal recessive cystic kidney disease that leads to renal failure in childhood or adolescence. Most NPHP gene products form molecular networks. Here we identify ANKS6 as a new NPHP family member that connects NEK8 (NPHP9) to INVS (NPHP2) and NPHP3. We show that ANKS6 localizes to the proximal cilium and confirm its role in renal development through knockdown experiments in zebrafish and Xenopus laevis. We also identify six families with ANKS6 mutations affected by nephronophthisis, including severe cardiovascular abnormalities, liver fibrosis and situs inversus. The oxygen sensor HIF1AN hydroxylates ANKS6 and INVS and alters the composition of the ANKS6-INVS-NPHP3 module. Knockdown of Hif1an in Xenopus results in a phenotype that resembles loss of other NPHP proteins. Network analyses uncovered additional putative NPHP proteins and placed ANKS6 at the center of this NPHP module, explaining the overlapping disease manifestation caused by mutation in ANKS6, NEK8, INVS or NPHP3.

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We are grateful to all patients and family members for their participation. We thank A. Sammarco, C. Engel, B. Müller, L. Schomas, S. Bräg and M. Klein for excellent technical assistance, the staff of the Life Imaging Center (LIC) in the Center for Systems Biology, Albert-Ludwigs-University Freiburg for excellent confocal microscopy resources and the support in image recording and analysis, and U. Lanner and E. Haaf of the proteomics core facility. We thank N. Katsanis and J. Willer (Duke University) for providing us with expression constructs for NEK7 and INVS. We thank K. Coene for her help in generating affinity proteomics data for NEK8. We thank E. Jones for making the 3G8 and 4A6 antibodies available through the European Xenopus stock centre. V.F., T.E., H.J.B. and C. Bergmann are employees of Bioscientia, a member of Sonic Healthcare. D.B. is a Higher Education Funding Council for England (HEFCE) Clinical Reader and is supported by Kids Kidney Research. A.K.-Z., G.W., E.W.K., F.G., T.B.H. and S.S.L. are supported by the Deutsche Forschungsgemeinschaft (DFG; KFO 201). E.W.K. is supported by the DFG (KU 1504). C. Boehlke is supported by the Else-Kröner-Fresenius Stiftung. G.W. and T.B.H. are supported by the Excellence Initiative of the German Federal and State Governments (EXC 294-BIOSS). M.U., R.R. and G.W. are supported by the European Community's Seventh Framework Programme (grant agreement 241955, SYSCILIA). R.R. is supported by the Netherlands Organisation for Scientific Research (NWO Vidi-91786396 and Vici-016.130.664). K.B. and M.U. are supported by the European Community's Seventh Framework Programme under grant agreement 278568, PRIMES. This study was supported in part by the Excellence Initiative of the German Federal and State Governments (GSC-4, Spemann Graduate School) and by grants from the Agence Nationale de la Recherche to S.S. (R09087KS and RPV11012KK) and the Fondation pour la Recherche Médicale (DEQ20071210558). This research was supported by grants from the US National Institutes of Health to F.H. (DK068306 and DK090917). F.H. is an Investigator of the Howard Hughes Medical Institute, a Doris Duke Distinguished Clinical Scientist and a Frederick G.L. Huetwell Professor. C. Bergmann received support from the DFG (BE 3910/4-1, ZE 205/14-1 and SFB/TRR57), the Deutsche Nierenstiftung and the PKD Foundation.

Author information

Author notes

    • Sylvia Hoff
    •  & Jan Halbritter

    These authors contributed equally to this work.


  1. Department of Medicine, Renal Division, University of Freiburg Medical Center, Freiburg, Germany.

    • Sylvia Hoff
    • , Daniel Epting
    • , Christopher Boehlke
    • , Christoph Schell
    • , Takayuki Yasunaga
    • , Martin Helmstädter
    • , Miriam Mergen
    • , Florian Grahammer
    • , Tobias B Huber
    • , E Wolfgang Kuehn
    • , Albrecht Kramer-Zucker
    • , Gerd Walz
    • , Carsten Bergmann
    •  & Soeren S Lienkamp
  2. Spemann Graduate School of Biology and Medicine (SGBM), Albert-Ludwigs-University Freiburg, Freiburg, Germany.

    • Sylvia Hoff
    •  & Christoph Schell
  3. Faculty of Biology, Albert-Ludwigs-University Freiburg, Freiburg, Germany.

    • Sylvia Hoff
    • , Christoph Schell
    •  & Martin Helmstädter
  4. Department of Medicine, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA.

    • Jan Halbritter
    •  & Friedhelm Hildebrandt
  5. Center for Human Genetics, Bioscientia, Ingelheim, Germany.

    • Valeska Frank
    • , Tobias Eisenberger
    • , Hanno J Bolz
    •  & Carsten Bergmann
  6. Department of Human Genetics, Nijmegen Centre for Molecular Life Sciences and Institute for Genetic and Metabolic Disease, Radboud University Medical Centre, Nijmegen, The Netherlands.

    • Thanh-Minh T Nguyen
    • , Jeroen van Reeuwijk
    •  & Ronald Roepman
  7. Institute National de la Santé et de la Recherche Médicale (INSERM) U983, Hôpital Necker–Enfants Malades, Paris, France.

    • Emilie Filhol
    •  & Sophie Saunier
  8. Paris Descartes–Sorbonne Paris Cité University, Imagine Institute, Paris, France.

    • Emilie Filhol
    •  & Sophie Saunier
  9. Institute for Ophthalmic Research, Division of Experimental Ophthalmology and Medical Proteome Center, University of Tuebingen, Tuebingen, Germany.

    • Karsten Boldt
    • , Nicola Horn
    •  & Marius Ueffing
  10. Research Unit Protein Science, Helmholtz Zentrum München–German Research Center for Environmental Health, Munich and Neuherberg, Germany.

    • Marius Ueffing
  11. Department of Pediatrics, University of Michigan, Ann Arbor, Michigan, USA.

    • Edgar A Otto
  12. Department of Clinical Genetics, VU University Medical Center, Amsterdam, The Netherlands.

    • Mariet W Elting
  13. Department of Pediatric Nephrology, VU University Medical Center, Amsterdam, The Netherlands.

    • Joanna A E van Wijk
  14. University College London (UCL) Institute of Child Health and Pediatric Nephrology, Great Ormond Street Hospital, London, UK.

    • Detlef Bockenhauer
  15. Department of Histopathology, Great Ormond Street Hospital, London, UK.

    • Neil J Sebire
  16. Department of Pediatrics, Aarhus University Hospital, Skejby, Aarhus, Denmark.

    • Søren Rittig
  17. Institute of Pathology, Aalborg University Hospital, Aalborg, Denmark.

    • Mogens Vyberg
  18. Department of Nephrology, Aalborg University Hospital, Aalborg, Denmark.

    • Troels Ring
  19. Department of Pediatrics and Adolescent Medicine, University of Freiburg Medical Center, Freiburg, Germany.

    • Martin Pohl
  20. Department of Pediatric Nephrology, Hannover Medical School, Hannover, Germany.

    • Lars Pape
  21. Department of Pediatrics, Children's Hospital Lucerne, Lucerne, Switzerland.

    • Thomas J Neuhaus
  22. Center of Pediatric Nephrology and Transplantation, Cairo University and Egyptian Group for Orphan Renal Diseases (EGORD), Cairo, Egypt.

    • Neveen A Soliman Elshakhs
  23. Division of Nephrology and Hypertension, Mayo Clinic, Rochester, Minnesota, USA.

    • Sarah J Koon
    •  & Peter C Harris
  24. Center for Biological Signaling Studies (BIOSS), Freiburg, Germany.

    • Tobias B Huber
    • , E Wolfgang Kuehn
    •  & Gerd Walz
  25. Institute of Human Genetics, University Hospital of Cologne, Cologne, Germany.

    • Hanno J Bolz
  26. Howard Hughes Medical Institute, Chevy Chase, Maryland, USA.

    • Friedhelm Hildebrandt
  27. Center for Clinical Research, University of Freiburg, Freiburg, Germany.

    • Carsten Bergmann


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S.H. performed Xenopus and biochemical experiments. D.E. performed zebrafish studies. C. Boehlke, C.S., T.Y., M.H. and M.M. analyzed cilia in various models. J.H., E.F., E.A.O., V.F., T.E., H.J.B., S.S., F.H. and C. Bergmann performed mutational analysis. J.v.R., T.-M.T.N., K.B., N.H., M.U. and R.R. performed affinity proteomic and network analyses. M.W.E., J.A.E.v.W., D.B., N.J.S., S.R., M.V., T.R., M.P., L.P., T.J.N., N.A.S.E., S.J.K. and P.C.H. recruited subjects and provided clinical information. S.H., D.E., T.Y., F.G., T.B.H., E.W.K., A.K.-Z., G.W. and S.S.L. designed experiments and analyzed data. S.H., J.H., R.R., S.S., C. Bergmann, F.H., G.W. and S.S.L. wrote the manuscript, with input from all authors.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Friedhelm Hildebrandt or Carsten Bergmann or Soeren S Lienkamp.

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