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

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 sev...

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Autores principales: Grampa, Valentina, Delous, Marion, Zaidan, Mohamad, Odye, Gweltas, Thomas, Sophie, Elkhartoufi, Nadia, Filhol, Emilie, Niel, Olivier, Silbermann, Flora, Lebreton, Corinne, Collardeau-Frachon, Sophie, Rouvet, Isabelle, Alessandri, Jean-Luc, Devisme, Louise, Dieux-Coeslier, Anne, Cordier, Marie-Pierre, Capri, Yline, Khung-Savatovsky, Suonavy, Sigaudy, Sabine, Salomon, Rémi, Antignac, Corinne, Gubler, Marie-Claire, Benmerah, Alexandre, Terzi, Fabiola, Attié-Bitach, Tania, Jeanpierre, Cécile, Saunier, Sophie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4788435/
https://www.ncbi.nlm.nih.gov/pubmed/26967905
http://dx.doi.org/10.1371/journal.pgen.1005894
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author Grampa, Valentina
Delous, Marion
Zaidan, Mohamad
Odye, Gweltas
Thomas, Sophie
Elkhartoufi, Nadia
Filhol, Emilie
Niel, Olivier
Silbermann, Flora
Lebreton, Corinne
Collardeau-Frachon, Sophie
Rouvet, Isabelle
Alessandri, Jean-Luc
Devisme, Louise
Dieux-Coeslier, Anne
Cordier, Marie-Pierre
Capri, Yline
Khung-Savatovsky, Suonavy
Sigaudy, Sabine
Salomon, Rémi
Antignac, Corinne
Gubler, Marie-Claire
Benmerah, Alexandre
Terzi, Fabiola
Attié-Bitach, Tania
Jeanpierre, Cécile
Saunier, Sophie
author_facet Grampa, Valentina
Delous, Marion
Zaidan, Mohamad
Odye, Gweltas
Thomas, Sophie
Elkhartoufi, Nadia
Filhol, Emilie
Niel, Olivier
Silbermann, Flora
Lebreton, Corinne
Collardeau-Frachon, Sophie
Rouvet, Isabelle
Alessandri, Jean-Luc
Devisme, Louise
Dieux-Coeslier, Anne
Cordier, Marie-Pierre
Capri, Yline
Khung-Savatovsky, Suonavy
Sigaudy, Sabine
Salomon, Rémi
Antignac, Corinne
Gubler, Marie-Claire
Benmerah, Alexandre
Terzi, Fabiola
Attié-Bitach, Tania
Jeanpierre, Cécile
Saunier, Sophie
author_sort Grampa, Valentina
collection PubMed
description 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.
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spelling pubmed-47884352016-03-23 Novel NEK8 Mutations Cause Severe Syndromic Renal Cystic Dysplasia through YAP Dysregulation Grampa, Valentina Delous, Marion Zaidan, Mohamad Odye, Gweltas Thomas, Sophie Elkhartoufi, Nadia Filhol, Emilie Niel, Olivier Silbermann, Flora Lebreton, Corinne Collardeau-Frachon, Sophie Rouvet, Isabelle Alessandri, Jean-Luc Devisme, Louise Dieux-Coeslier, Anne Cordier, Marie-Pierre Capri, Yline Khung-Savatovsky, Suonavy Sigaudy, Sabine Salomon, Rémi Antignac, Corinne Gubler, Marie-Claire Benmerah, Alexandre Terzi, Fabiola Attié-Bitach, Tania Jeanpierre, Cécile Saunier, Sophie PLoS Genet Research Article 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. Public Library of Science 2016-03-11 /pmc/articles/PMC4788435/ /pubmed/26967905 http://dx.doi.org/10.1371/journal.pgen.1005894 Text en © 2016 Grampa et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Grampa, Valentina
Delous, Marion
Zaidan, Mohamad
Odye, Gweltas
Thomas, Sophie
Elkhartoufi, Nadia
Filhol, Emilie
Niel, Olivier
Silbermann, Flora
Lebreton, Corinne
Collardeau-Frachon, Sophie
Rouvet, Isabelle
Alessandri, Jean-Luc
Devisme, Louise
Dieux-Coeslier, Anne
Cordier, Marie-Pierre
Capri, Yline
Khung-Savatovsky, Suonavy
Sigaudy, Sabine
Salomon, Rémi
Antignac, Corinne
Gubler, Marie-Claire
Benmerah, Alexandre
Terzi, Fabiola
Attié-Bitach, Tania
Jeanpierre, Cécile
Saunier, Sophie
Novel NEK8 Mutations Cause Severe Syndromic Renal Cystic Dysplasia through YAP Dysregulation
title Novel NEK8 Mutations Cause Severe Syndromic Renal Cystic Dysplasia through YAP Dysregulation
title_full Novel NEK8 Mutations Cause Severe Syndromic Renal Cystic Dysplasia through YAP Dysregulation
title_fullStr Novel NEK8 Mutations Cause Severe Syndromic Renal Cystic Dysplasia through YAP Dysregulation
title_full_unstemmed Novel NEK8 Mutations Cause Severe Syndromic Renal Cystic Dysplasia through YAP Dysregulation
title_short Novel NEK8 Mutations Cause Severe Syndromic Renal Cystic Dysplasia through YAP Dysregulation
title_sort novel nek8 mutations cause severe syndromic renal cystic dysplasia through yap dysregulation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4788435/
https://www.ncbi.nlm.nih.gov/pubmed/26967905
http://dx.doi.org/10.1371/journal.pgen.1005894
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