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Mutations in the Spliceosome Component CWC27 Cause Retinal Degeneration with or without Additional Developmental Anomalies

Apr 5, 2017

Mingchu Xu, Yajing (Angela) Xie, Hana Abouzeid, Christopher T Gordon, Alessia Fiorentino, Zixi Sun, Anna Lehman, Ihab S Osman, Rachayata Dharmat, Rosa Riveiro-Alvarez, Linda Bapst-Wicht, Darwin Babino, Gavin Arno, Virginia Busetto, Li Zhao, Hui Li, Miguel A Lopez-Martinez, Liliana F Azevedo, Laurence Hubert, Nikolas Pontikos, Aiden Eblimit, Isabel Lorda-Sanchez, Valeria Kheir, Vincent Plagnol, Myriam Oufadem, Zachry T Soens, Lizhu Yang, Christine Bole-Feysot, Rolph Pfundt, Nathalie Allaman-Pillet, Patrick Nitschké, Michael E Cheetham, Stanislas Lyonnet, Smriti A Agrawal, Huajin Li, Gaëtan Pinton, Michel Michaelides, Claude Besmond, Yumei Li, Zhisheng Yuan, Johannes von Lintig, Andrew R Webster, Hervé Le Hir, Peter Stoilov, Jeanne Amiel, Alison J Hardcastle, Carmen Ayuso, Ruifang Sui, Rui Chen, Rando Allikmets, Daniel F Schorderet | American Journal Human Genetics | 2017 Apr 6 | 100(4) | pgs. 592-604 | doi: 10.1016/j.ajhg.2017.02.008


Abstract

Pre-mRNA splicing factors play a fundamental role in regulating transcript diversity both temporally and spatially. Genetic defects in several spliceosome components have been linked to a set of non-overlapping spliceosomopathy phenotypes in humans, among which skeletal developmental defects and non-syndromic retinitis pigmentosa (RP) are frequent findings. Here we report that defects in spliceosome-associated protein CWC27 are associated with a spectrum of disease phenotypes ranging from isolated RP to severe syndromic forms. By whole-exome sequencing, recessive protein-truncating mutations in CWC27 were found in seven unrelated families that show a range of clinical phenotypes, including retinal degeneration, brachydactyly, craniofacial abnormalities, short stature, and neurological defects. Remarkably, variable expressivity of the human phenotype can be recapitulated in Cwc27 mutant mouse models, with significant embryonic lethality and severe phenotypes in the complete knockout mice while mice with a partial loss-of-function allele mimic the isolated retinal degeneration phenotype. Our study describes a retinal dystrophy-related phenotype spectrum as well as its genetic etiology and highlights the complexity of the spliceosomal gene network.


Introduction

Pre-mRNA splicing, which removes introns from eukaryotic transcripts, is an essential step in gene expression. Through the generation of numerous alternatively spliced transcript isoforms from the limited set of genes, the splicing process plays a critical role in giving rise to the protein diversity necessary to establish the complex structures and functions found throughout eukaryotes.1, 2 Splicing of pre-mRNA is catalyzed by the spliceosome, a ribonucleoprotein (RNP) complex that is dynamically assembled on each intron and undergoes several rearrangement steps before excising the intron.3 The core of the spliceosome is formed by five small nuclear RNP (snRNP) particles and proteomic studies have identified more than 150 spliceosomal proteins including snRNP-specific proteins as well as miscellaneous non-snRNP splicing factors.4, 5, 6, 7 Though expressed ubiquitously, most spliceosomal genes associated with Mendelian disease have been classified within one of two non-overlapping phenotypic groups, suggesting tissue-specific functional roles. Mutations in splicing factors TXNL4A (MIM: 611595),8 RBM8A (MIM: 605313),9 SNRPB (MIM: 182282),10 EIF4A3 (MIM: 608546),11 EFTUD2 (MIM: 603892),12 and SF3B4 (MIM: 605593)13 cause syndromes mainly involving craniofacial and skeletal abnormalities, while disruptions of another group of spliceosomal genes—PRPF3 (MIM: 607301),14 PRPF31 (MIM: 606419),15 PRPF4 (MIM: 607795),16 PRPF6 (MIM: 613979),17 PRPF8 (MIM: 607300),18 and SNRNP200 (MIM: 601664)19—lead to non-syndromic retinitis pigmentosa (RP), a restricted disease phenotype primarily affecting the rod photoreceptors. Recent next-generation sequencing approaches have allowed the identification of many of the disease-associated splicing factors listed above, but nevertheless the structural and functional roles of most spliceosome components and their involvement in human disease remain elusive.20


Here, by exome sequencing in multiple families and disease modeling of two mouse alleles, we show that the disruption of the spliceosomal gene CWC27 (MIM: 617170) leads to a spectrum of isolated to syndromic phenotypes. The syndrome features include retinal degeneration, brachydactyly, craniofacial abnormalities, short stature, and neurological defects, with convergence of the two aforementioned non-overlapping spliceosomopathy phenotype groups. This study identifies a role for CWC27 both during early development and in the maintenance of mature tissues and highlights the complexity of spliceosome function.


 

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