Mutations in GREB1L Cause Bilateral Kidney Agenesis in Humans and Mice

Lara de Tomasi (1, 2) , Pierre David (3) , Camille Humbert (1) , Flora Silbermann (1) , Christelle Arrondel (1) , Frédéric Tores (4) , Stéphane Fouquet (5) , Audrey Desgrange (6, 7) , Olivier Niel (8) , Christine Bole-Feysot (4) , Patrick Nitschké (4) , Joëlle Roume (9) , Marie-Pierre Cordier , Christine Pietrement (10) , Bertrand Isidor (11) , Philippe Khau van Kien (12) , Marie Gonzales (13) , Marie-Hélène Saint-Frison , Jelena Martinovic (14) , Robert Novo (15) , Juliette Piard , Christelle Cabrol (16) , Ishwar Verma (16) , Ratna Puri (17) , Hubert Journel , Jacqueline Aziza , Laurent Gavard , Marie-Hélène Said-Menthon , Laurence Heidet , Sophie Saunier (18) , Cécile Jeanpierre (18)

1 Equipe Inserm U1163 - The molecular basis of kidney diseases: cystinosis and hereditary nephrotic syndrome
2 UPD7 - Université Paris Diderot - Paris 7
3 SFR Necker - UMS 3633 / US24 - Structure Fédérative de Recherche Necker
4 Imagine - U1163 - Imagine - Institut des maladies génétiques (IHU)
5 Institut de la Vision
6 Imagine - Institut Pasteur U1163 - Coordination des Cellules et Morphogenèse / Heart Morphogenesis
7 UPD5 - Université Paris Descartes - Paris 5
8 Hôpital Robert Debré Paris
9 CHI Poissy-Saint-Germain
10 CHU Reims - Centre Hospitalier Universitaire de Reims
11 CHU Nantes - Centre hospitalier universitaire de Nantes
12 CHU Nîmes - Hôpital Universitaire Carémeau [Nîmes]
13 Laboratoire Histologie Embryologie Cytogénétique [CHU Necker]
14 AP-HP - Hôpital Antoine Béclère [Clamart]
15 CHU Lille
16 CHRU Besançon - Centre Hospitalier Régional Universitaire de Besançon
17 Sir Ganga Ram Hospital [New Delhi, India]
18 CHU Necker - Enfants Malades [AP-HP]

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

Domains: Morphogenesis

Lara de Tomasi, Pierre David, Camille Humbert, Flora Silbermann, Christelle Arrondel, et al.. Mutations in GREB1L Cause Bilateral Kidney Agenesis in Humans and Mice. American Journal of Human Genetics, 2017, 101 (5), pp.803-814. ⟨10.1016/j.ajhg.2017.09.026⟩. ⟨pasteur-03931320⟩