ETV4 Mutation in a Patient with Congenital Anomalies of the Kidney and Urinary Tract

(Pages 61-71)
Jing Chen1, 2, #, Amelie T. van der Ven1, #, Joseph A. Newman3, Asaf Vivante1, 4, Nina Mann1, Hazel Aitkenhead3, Shirlee Shril1, Hadas Ityel1, Julian Schulz1, Johanna Magdalena Schmidt1, Eugen Widmeier1, 5, Opher Gileadi3, Frank Costantini6, Shifaan Thowfeequ6, Roland H. Wenger7, Stuart B. Bauer8, Richard S. Lee8, Weining Lu9, Maike Getwan5, Michael M. Kaminski5, Soeren S. Lienkamp5, 10, Richard P. Lifton11, 12, Velibor Tasic13, Elijah O. Kehinde14 and Friedhelm Hildebrandt1

1Department of Medicine, Boston Children’s Hospital, Harvard Medical School, Boston, Massachusetts, USA; 2Department of Nephrology, Children’s Hospital of Fudan University, Shanghai, China; 3The Structural Genomics Consortium, University of Oxford, Old Road Campus Research Building, Oxford, United Kingdom; 4Talpiot Medical Leadership Program, Sheba Medical Center, Tel-Hashomer, Israel; 5Department of Medicine, Renal Division, Medical Center – University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany; 6Department of Genetics and Development, Columbia University Medical Center, New York, New York, USA; 7Institute of Physiology and Zürich Center for Integrative Human Physiology (ZIHP), University of Zürich, Zürich, Switzerland; 8Department of Urology, Boston Children’s Hospital, Harvard Medical School, Massachusetts, USA; 9Renal Section, Department of Medicine, Boston University Medical Center, Boston, MA, USA; 10Center for Biological Signaling Studies (BIOSS), Freiburg, Germany; 11Department of Human Genetics, Yale University School of Medicine, New Haven, CT, USA; 12Howard Hughes Medical Institute, Chevy Chase, Maryland, USA; 13Medical Faculty Skopje, University Children’s Hospital, Skopje, Macedonia; 14Division of Urology, Department of Surgery, Nazarbayev University, Astana, Kazakhstan



Abstract: Congenital anomalies of the kidney and urinary tract (CAKUT) are the most common reason for chronic kidney disease in children. Although more than 30 monogenic causes have been implicated in isolated forms of human CAKUT so far, the vast majority remains elusive. To identify novel monogenic causes of CAKUT we applied homozygosity mapping, together with whole exome sequencing, in a patient from consanguineous descent with isolated CAKUT. We identified a homozygous missense mutation (p.Arg415His) of the Ets Translocation Variant Gene 4 (ETV4). The transcription factor ETV4 is a downstream target of the GDNF/RET signaling pathway that plays a crucial role in kidney development. We show by means of electrophoretic mobility shift assay that the Arg415His mutant causes loss of the DNA binding affinity of ETV4 and fails to activate transcription in a cell-based luciferase reporter assay. We furthermore investigated the impact of the mutant protein on cell migration rate. Unlike wildtype ETV4, the Arg415His mutant failed to rescue cell migration defects observed in two ETV4 knock-down cell-lines. We therefore identified and functionally characterized a recessive mutation in ETV4 in a human patient with CAKUT. We hypothesize that the pathomechanism of this mutation could be via loss of the transcriptional function of ETV4, and a resulting abrogation of GDNF/RET/ETV4 signaling pathway.

Keywords: CAKUT, cell migration, DNA binding, ETS domain.