PUBLICATION

Novel NEK8 Mutations Cause Severe Syndromic Renal Cystic Dysplasia through YAP Dysregulation

Authors

Grampa, V., Delous, M., Zaidan, M., Odye, G., Thomas, S., Elkhartoufi, N., Filhol, E., Niel, O., Silbermann, F., Lebreton, C., Collardeau-Frachon, S., Rouvet, I., Alessandri, J.L., Devisme, L., Dieux-Coeslier, A., Cordier, M.P., Capri, Y., Khung-Savatovsky, S., Sigaudy, S., Salomon, R., Antignac, C., Gubler, M.C., Benmerah, A., Terzi, F., Attié-Bitach, T., Jeanpierre, C., Saunier, S.

ID

ZDB-PUB-160312-1

Date

2016

Source

PLoS Genetics 12: e1005894 (Journal)

Registered Authors

Delous, Marion, Saunier, Sophie

Keywords

Fibroblasts, Cilia, Kidneys, Nonsense mutation, Cell staining, Embryos, Missense mutation, Nuclear staining

MeSH Terms

Adaptor Proteins, Signal Transducing/antagonists & inhibitors
Adaptor Proteins, Signal Transducing/biosynthesis
Adaptor Proteins, Signal Transducing/genetics*
Animals
Cell Differentiation/genetics
Cilia/genetics*
Cilia/pathology
Female
Genetic Association Studies
Humans
Kidney/metabolism
Kidney/pathology
Mice
Morphogenesis/genetics
Mutation
NIMA-Related Kinases
Phosphoproteins/antagonists & inhibitors
Phosphoproteins/biosynthesis
Phosphoproteins/genetics*
Polycystic Kidney Diseases/genetics*
Polycystic Kidney Diseases/pathology
Porphyrins/administration & dosage
Protein Kinases/genetics*
Signal Transduction
Zebrafish

PubMed

26967905 Full text @ PLoS Genet.

Abstract

Ciliopathies are a group of genetic multi-systemic disorders related to dysfunction of the primary cilium, a sensory organelle present at the cell surface that regulates key signaling pathways during development and tissue homeostasis. In order to identify novel genes whose mutations would cause severe developmental ciliopathies, >500 patients/fetuses were analyzed by a targeted high throughput sequencing approach allowing exome sequencing of >1200 ciliary genes. NEK8/NPHP9 mutations were identified in five cases with severe overlapping phenotypes including renal cystic dysplasia/hypodysplasia, situs inversus, cardiopathy with hypertrophic septum and bile duct paucity. These cases highlight a genotype-phenotype correlation, with missense and nonsense mutations associated with hypodysplasia and enlarged cystic organs, respectively. Functional analyses of NEK8 mutations in patient fibroblasts and mIMCD3 cells showed that these mutations differentially affect ciliogenesis, proliferation/apoptosis/DNA damage response, as well as epithelial morphogenesis. Notably, missense mutations exacerbated some of the defects due to NEK8 loss of function, highlighting their likely gain-of-function effect. We also showed that NEK8 missense and loss-of-function mutations differentially affect the regulation of the main Hippo signaling effector, YAP, as well as the expression of its target genes in patient fibroblasts and renal cells. YAP imbalance was also observed in enlarged spheroids of Nek8-invalidated renal epithelial cells grown in 3D culture, as well as in cystic kidneys of Jck mice. Moreover, co-injection of nek8 MO with WT or mutated NEK8-GFP RNA in zebrafish embryos led to shortened dorsally curved body axis, similar to embryos injected with human YAP RNA. Finally, treatment with Verteporfin, an inhibitor of YAP transcriptional activity, partially rescued the 3D spheroid defects of Nek8-invalidated cells and the abnormalities of NEK8-overexpressing zebrafish embryos. Altogether, our study demonstrates that NEK8 human mutations cause major organ developmental defects due to altered ciliogenesis and cell differentiation/proliferation through deregulation of the Hippo pathway.

Genes / Markers

Figures

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Expression

Phenotype

Mutations / Transgenics

Human Disease / Model

Sequence Targeting Reagents

Fish

Antibodies

Orthology

Engineered Foreign Genes

Mapping

Grampa et al., 2016 ZDB-PUB-160312-1 PMID:26967905

Novel NEK8 Mutations Cause Severe Syndromic Renal Cystic Dysplasia through YAP Dysregulation

Grampa et al., 2016

ZDB-PUB-160312-1
PMID:26967905