



@article {3267,
	title = {Rescue of wild-type E-cadherin expression from nonsense-mutated cancer cells by a suppressor-tRNA},
	journal = {European Journal of Human Genetics},
	volume = {22},
	year = {2014},
	note = {Export Date: 21 November 2014},
	month = {2014},
	pages = {1085 - 1092},
	abstract = {Hereditary diffuse gastric cancer (HDGC) syndrome, although rare, is highly penetrant at an early age, and is severe and incurable because of ineffective screening tools and therapy. Approximately 45\% of HDGC families carry germline CDH1/E-cadherin alterations, 20\% of which are nonsense leading to premature protein truncation. Prophylactic gastrectomy is the only recommended approach for all asymptomatic CDH1 mutation carriers. Suppressor-tRNAs can replace premature stop codons (PTCs) with a cognate amino acid, inducing readthrough and generating full-length proteins. The use of suppressor-tRNAs in HDGC patients could therefore constitute a less invasive therapeutic option for nonsense mutation carriers, delaying the development of gastric cancer. Our analysis revealed that 23/108 (21.3\%) of E-cadherin-mutant families carried nonsense mutations that could be potentially corrected by eight suppressor-tRNAs, and arginine was the most frequently affected amino acid. Using site-directed mutagenesis, we developed an arginine suppressor-tRNA vector to correct one HDGC nonsense mutation. E-cadherin-deficient cell lines were transfected with plasmids carrying simultaneously the suppressor-tRNA and wild-type or mutant CDH1 mini-genes. RT-PCR, western blot, immunofluorescence, flow cytometry and proximity ligation assay (PLA) were used to establish the model, and monitor mRNA and protein expression and function recovery from CDH1 vectors. Cells expressing a CDH1 mini-gene, carrying a nonsense mutation and the suppressor-tRNA, recovered full-length E-cadherin expression and its correct localization and incorporation into the adhesion complex. This is the first demonstration of functional recovery of a mutated causative gene in hereditary cancer by cognate amino acid replacement with suppressor-tRNAs. Of the HDGC families, 21.3\% are candidates for correction with suppressor-tRNAs to potentially delay cancer onset. {\textcopyright} 2014 Macmillan Publishers Limited. All rights reserved.},
	keywords = {hereditary diffuse gastric cancer (HDGC), nonsense suppression, Premature termination codon (PTC), Suppressor-tRNA},
	url = {http://www.scopus.com/inward/record.url?eid=2-s2.0-84906269658\&partnerID=40\&md5=2feed844660a13385857a7e5d4470a2b},
	author = {Bordeira-Carri{\c c}o, R. and Ferreira, D. and Mateus, D.D. and Pinheiro, H. and P{\^e}go, A.P. and Santos, M.A.S. and Oliveira, C.}
}
