Cnbp ameliorates Treacher Collins Syndrome craniofacial anomalies through a pathway that involves redox-responsive genes.

Mauro S Porcel De Peralta,Gabriela Coux,Roberto D Fanganiello,Nora B Calcaterra,Valeria S Mouguelar,Maria R Passos-Bueno,María Antonella Sdrigotti,Felipe A A Ishiy

doi:10.1038/cddis.2016.299

Abstract

Treacher Collins Syndrome (TCS) is a rare congenital disease (1:50 000 live births) characterized by craniofacial defects, including hypoplasia of facial bones, cleft palate and palpebral fissures. Over 90% of the cases are due to mutations in the TCOF1 gene, which codifies the nucleolar protein Treacle. Here we report a novel TCS-like zebrafish model displaying features that fully recapitulate the spectrum of craniofacial abnormalities observed in patients. As it was reported for a Tcof1+/− mouse model, Treacle depletion in zebrafish caused reduced rRNA transcription, stabilization of Tp53 and increased cell death in the cephalic region. An increase of ROS along with the overexpression of redox-responsive genes was detected; furthermore, treatment with antioxidants ameliorated the phenotypic defects of craniofacial anomalies in TCS-like larvae. On the other hand, Treacle depletion led to a lowering in the abundance of Cnbp, a protein required for proper craniofacial development. Tcof1 knockdown in transgenic zebrafish overexpressing cnbp resulted in barely affected craniofacial cartilage development, reinforcing the notion that Cnbp has a role in the pathogenesis of TCS. The cnbp overexpression rescued the TCS phenotype in a dose-dependent manner by a ROS-cytoprotective action that prevented the redox-responsive genes' upregulation but did not normalize the synthesis of rRNAs. Finally, a positive correlation between the expression of CNBP and TCOF1 in mesenchymal cells from both control and TCS subjects was found. Based on this, we suggest CNBP as an additional target for new alternative therapeutic treatments to reduce craniofacial defects not only in TCS but also in other neurocristopathies.

Highlights

Treacher Collins Syndrome (TCS) (Online Mendelian Inheritance in Man database accession no. 154500) is a congenital craniofacial disorder being characterized by numerous anomalies, restricted primarily to the head and neck
The current hypothesis states that TCS phenotypes are due to an aberrant ribosome biogenesis within neuroepithelial cells, enhancing apoptosis mediated by Tp53stabilization.[10,15]
Ribosomal DNA transcription is one of the limiting steps of ribosome biogenesis.[22]. Thereby, we explored this issue by directly measuring by real-time quantitative PCR (RT-qPCR) of the 47S ribosomal RNA (rRNA) abundance in 256-cell stage and 24 hours postfertilization STD- and TRA-morphants

Summary

Introduction

Treacher Collins Syndrome (TCS) (Online Mendelian Inheritance in Man database accession no. 154500) is a congenital craniofacial disorder being characterized by numerous anomalies, restricted primarily to the head and neck. The phenotype of TCS includes hypoplasia of the facial bones, the zygomatic complex and mandible, cleft palate and middle and external ear defects that result in conductive deafness.[1] Many of the tissues affected in the syndrome arise from the first (maxillary and mandibular) and second (hyoid) pharyngeal arches.[1] These arches are strongly colonized by cranial neural crest cells (CNCCs), which are migratory multipotent progenitor cells derived from the neuroepithelium They form much of the cartilage, bone and connective tissues of the head and the face.[2]. (formerly nolc1-like) expression was restricted to the anteriormost region of zebrafish developing embryos,[14] to Received 26.4.16; revised 28.7.16; accepted 25.8.16; Edited by A Finazzi-Agro'

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