Mutations in GREB1L Cause Bilateral Kidney Agenesis in Humans and Mice

  • Am J Hum Genet. 2017 Nov 2;101(5):803-814. doi: 10.1016/j.ajhg.2017.09.026.
Lara De Tomasi  1 ,  Pierre David  2 ,  Camille Humbert  3 ,  Flora Silbermann  3 ,  Christelle Arrondel  3 ,  Frédéric Tores  4 ,  Stéphane Fouquet  5 ,  Audrey Desgrange  6 ,  Olivier Niel  7 ,  Christine Bole-Feysot  8 ,  Patrick Nitschké  4 ,  Joëlle Roume  9 ,  Marie-Pierre Cordier  10 ,  Christine Pietrement  11 ,  Bertrand Isidor  12 ,  Philippe Khau Van Kien  13 ,  Marie Gonzales  14 ,  Marie-Hélène Saint-Frison  15 ,  Jelena Martinovic  16 ,  Robert Novo  17 ,  Juliette Piard  18 ,  Christelle Cabrol  18 ,  Ishwar C Verma  19 ,  Ratna Puri  19 ,  Hubert Journel  20 ,  Jacqueline Aziza  21 ,  Laurent Gavard  22 ,  Marie-Hélène Said-Menthon  23 ,  Laurence Heidet  24 ,  Sophie Saunier  3 ,  Cécile Jeanpierre  25
Affiliations
  • 1. Laboratory of Hereditary Kidney Diseases, INSERM UMR 1163, Imagine Institute, 75015 Paris, France; Paris Descartes-Sorbonne Paris Cité University, Imagine Institute, 75015 Paris, France; Paris Diderot University, 75013 Paris, France.
  • 2. Transgenesis Platform, Laboratoire d'Expérimentation Animale et Transgenèse (LEAT), Imagine Institute, Structure Fédérative de Recherche Necker INSERM US24/CNRS UMS3633, 75015 Paris, France.
  • 3. Laboratory of Hereditary Kidney Diseases, INSERM UMR 1163, Imagine Institute, 75015 Paris, France; Paris Descartes-Sorbonne Paris Cité University, Imagine Institute, 75015 Paris, France.
  • 4. Bioinformatic Platform, INSERM UMR 1163, Paris Descartes-Sorbonne Paris Cité University, Imagine Institute, 75015 Paris, France.
  • 5. Imaging Platform, Sorbonne Universités, UPMC Univ Paris 06, INSERM UMR_S968 and CNRS UMR7210, Institut de la Vision, 75012 Paris, France.
  • 6. Paris Descartes-Sorbonne Paris Cité University, Imagine Institute, 75015 Paris, France; Laboratory of Heart Morphogenesis, INSERM UMR 1163, Imagine Institute-Pasteur Institute, 75015 Paris, France.
  • 7. APHP, Pediatric Nephrology Department, Hôpital Robert Debré, 75019 Paris, France.
  • 8. Genomic Platform, INSERM UMR 1163, Paris Descartes-Sorbonne Paris Cité University, Imagine Institute, 75015 Paris, France.
  • 9. Unité de Génétique Médicale, Centre de Référence des Maladies rares du Développement (AnD DI Rares), CHI Poissy - St Germain en Laye, 78300 Poissy, France.
  • 10. Service de Génétique, Groupement Hospitalier Est, 69677 Bron, France.
  • 11. Unité de Néphrologie Pédiatrique, CHU Reims, 51100 Reims, France.
  • 12. Service de Génétique Médicale, CHU Nantes, 44093 Nantes, France.
  • 13. Unité de Génétique Médicale, Nîmes University Hospital, CHU Carémeau, 30900 Nîmes, France.
  • 14. APHP, Département de Génétique Médicale, Hôpital Armand Trousseau, Université Pierre et Marie Curie, 75571 Paris, France and APHP, Unité d'EmbryoFœtopathologie, Service d'Histologie-Embryologie-Cytogénétique, Hôpital Necker-Enfants Malades, 75015 Paris, France.
  • 15. APHP, Département de Génétique, Unité de Fœtopathologie, Hôpital Robert Debré, 75019 Paris, France.
  • 16. APHP, Unit of Fetal Pathology, Antoine Béclère Hospital, 92140 Clamart, France.
  • 17. Centre Hospitalier Universitaire de Lille, Hôpital Jeanne de Flandre, Service de Néphrologie Pédiatrique, 59000 Lille, France.
  • 18. Centre de Génétique Humaine, CHU Besançon, Université de Franche-Comté, 25000 Besançon, France.
  • 19. Institute of Medical Genetics and Genomics, Sir Ganga Ram Hospital, New Delhi 110060, India.
  • 20. Service de Génétique Médicale, Hôpital Chubert, 56000 Vannes, France.
  • 21. Département d'Anatomie et Cytologie Pathologiques, IUCT-Oncopole, 31100 Toulouse, France.
  • 22. Service de Gynécologie-Obstétrique, Hôpital Louis Mourier, 92700 Colombes, France.
  • 23. Service de Pédiatrie, Hôpitaux du Léman, 74203 Thonon les Bains, France.
  • 24. APHP, Centre de Référence des Maladies Rénales Héréditaires de l'Enfant et de l'Adulte (MARHEA), Hôpital Necker-Enfants Malades, 75015 Paris, France; APHP, Service de Néphrologie Pédiatrique, Hôpital Necker-Enfants Malades, 75015 Paris, France.
  • 25. Laboratory of Hereditary Kidney Diseases, INSERM UMR 1163, Imagine Institute, 75015 Paris, France; Paris Descartes-Sorbonne Paris Cité University, Imagine Institute, 75015 Paris, France. Electronic address: [email protected].
PMID: 29100091 DOI: 10.1016/j.ajhg.2017.09.026
Abstract

Congenital anomalies of the kidney and urinary tract (CAKUT) constitute a major cause of chronic kidney disease in children and 20% of prenatally detected anomalies. CAKUT encompass a spectrum of developmental kidney defects, including renal agenesis, hypoplasia, and cystic and non-cystic dysplasia. More than 50 genes have been reported as mutated in CAKUT-affected case subjects. However, the pathophysiological mechanisms leading to bilateral kidney agenesis (BKA) remain largely elusive. Whole-exome or targeted exome Sequencing of 183 unrelated familial and/or severe CAKUT-affected case subjects, including 54 fetuses with BKA, led to the identification of 16 heterozygous variants in GREB1L (growth regulation by estrogen in breast Cancer 1-like), a gene reported as a target of retinoic acid signaling. Four loss-of-function and 12 damaging missense variants, 14 being absent from GnomAD, were identified. Twelve of them were present in familial or simplex BKA-affected case subjects. Female BKA-affected fetuses also displayed uterus agenesis. We demonstrated a significant association between GREB1L variants and BKA. By in situ hybridization, we showed expression of Greb1l in the nephrogenic zone in developing mouse kidney. We generated a Greb1l knock-out mouse model by CRISPR-Cas9. Analysis at E13.5 revealed lack of kidneys and genital tract anomalies in male and female Greb1l-/- embryos and a slight decrease in ureteric bud branching in Greb1l+/- embryos. We showed that Greb1l invalidation in mIMCD3 cells affected tubulomorphogenesis in 3D-collagen culture, a phenotype rescued by expression of the wild-type human protein. This demonstrates that GREB1L plays a major role in early metanephros and genital development in mice and humans.

Keywords
3DISCO; CAKUT; GREB1L; genital tract; heart defect; kidney agenesis; kidney development; mouse model; nephrogenesis; tubulogenesis.