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...
Saved in:
| Main Authors: | , , , , , , , , , , , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2016-03-01
|
| Series: | PLoS Genetics |
| Online Access: | https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1005894&type=printable |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850189237263007744 |
|---|---|
| author | Valentina Grampa Marion Delous Mohamad Zaidan Gweltas Odye Sophie Thomas Nadia Elkhartoufi Emilie Filhol Olivier Niel Flora Silbermann Corinne Lebreton Sophie Collardeau-Frachon Isabelle Rouvet Jean-Luc Alessandri Louise Devisme Anne Dieux-Coeslier Marie-Pierre Cordier Yline Capri Suonavy Khung-Savatovsky Sabine Sigaudy Rémi Salomon Corinne Antignac Marie-Claire Gubler Alexandre Benmerah Fabiola Terzi Tania Attié-Bitach Cécile Jeanpierre Sophie Saunier |
| author_facet | Valentina Grampa Marion Delous Mohamad Zaidan Gweltas Odye Sophie Thomas Nadia Elkhartoufi Emilie Filhol Olivier Niel Flora Silbermann Corinne Lebreton Sophie Collardeau-Frachon Isabelle Rouvet Jean-Luc Alessandri Louise Devisme Anne Dieux-Coeslier Marie-Pierre Cordier Yline Capri Suonavy Khung-Savatovsky Sabine Sigaudy Rémi Salomon Corinne Antignac Marie-Claire Gubler Alexandre Benmerah Fabiola Terzi Tania Attié-Bitach Cécile Jeanpierre Sophie Saunier |
| author_sort | Valentina Grampa |
| collection | DOAJ |
| 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. |
| format | Article |
| id | doaj-art-80b9228e4b7a4d9cafbccaa2a58ded5c |
| institution | OA Journals |
| issn | 1553-7390 1553-7404 |
| language | English |
| publishDate | 2016-03-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Genetics |
| spelling | doaj-art-80b9228e4b7a4d9cafbccaa2a58ded5c2025-08-20T02:15:40ZengPublic Library of Science (PLoS)PLoS Genetics1553-73901553-74042016-03-01123e100589410.1371/journal.pgen.1005894Novel NEK8 Mutations Cause Severe Syndromic Renal Cystic Dysplasia through YAP Dysregulation.Valentina GrampaMarion DelousMohamad ZaidanGweltas OdyeSophie ThomasNadia ElkhartoufiEmilie FilholOlivier NielFlora SilbermannCorinne LebretonSophie Collardeau-FrachonIsabelle RouvetJean-Luc AlessandriLouise DevismeAnne Dieux-CoeslierMarie-Pierre CordierYline CapriSuonavy Khung-SavatovskySabine SigaudyRémi SalomonCorinne AntignacMarie-Claire GublerAlexandre BenmerahFabiola TerziTania Attié-BitachCécile JeanpierreSophie SaunierCiliopathies 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.https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1005894&type=printable |
| spellingShingle | Valentina Grampa Marion Delous Mohamad Zaidan Gweltas Odye Sophie Thomas Nadia Elkhartoufi Emilie Filhol Olivier Niel Flora Silbermann Corinne Lebreton Sophie Collardeau-Frachon Isabelle Rouvet Jean-Luc Alessandri Louise Devisme Anne Dieux-Coeslier Marie-Pierre Cordier Yline Capri Suonavy Khung-Savatovsky Sabine Sigaudy Rémi Salomon Corinne Antignac Marie-Claire Gubler Alexandre Benmerah Fabiola Terzi Tania Attié-Bitach Cécile Jeanpierre Sophie Saunier Novel NEK8 Mutations Cause Severe Syndromic Renal Cystic Dysplasia through YAP Dysregulation. PLoS Genetics |
| 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 |
| url | https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1005894&type=printable |
| work_keys_str_mv | AT valentinagrampa novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT mariondelous novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT mohamadzaidan novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT gweltasodye novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT sophiethomas novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT nadiaelkhartoufi novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT emiliefilhol novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT olivierniel novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT florasilbermann novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT corinnelebreton novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT sophiecollardeaufrachon novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT isabellerouvet novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT jeanlucalessandri novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT louisedevisme novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT annedieuxcoeslier novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT mariepierrecordier novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT ylinecapri novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT suonavykhungsavatovsky novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT sabinesigaudy novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT remisalomon novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT corinneantignac novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT marieclairegubler novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT alexandrebenmerah novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT fabiolaterzi novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT taniaattiebitach novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT cecilejeanpierre novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation AT sophiesaunier novelnek8mutationscauseseveresyndromicrenalcysticdysplasiathroughyapdysregulation |