TY - JOUR
T1 - Gain-of-function mutations in DNMT3A in patients with paraganglioma
AU - Remacha, Laura
AU - Currás-Freixes, Maria
AU - Torres-Ruiz, Raúl
AU - Schiavi, Francesca
AU - Torres-Pérez, Rafael
AU - Calsina, Bruna
AU - Letón, Rocío
AU - Comino-Méndez, Iñaki
AU - Roldán-Romero, Juan M
AU - Montero-Conde, Cristina
AU - Santos, María
AU - Pérez, Lucía Inglada
AU - Pita, Guillermo
AU - Alonso, María R
AU - Honrado, Emiliano
AU - Pedrinaci, Susana
AU - Crespo-Facorro, Benedicto
AU - Percesepe, Antonio
AU - Falcioni, Maurizio
AU - Rodríguez-Perales, Sandra
AU - Korpershoek, Esther
AU - Ramón-Maiques, Santiago
AU - Opocher, Giuseppe
AU - Rodríguez-Antona, Cristina
AU - Robledo, Mercedes
AU - Cascón, Alberto
PY - 2018/5/8
Y1 - 2018/5/8
N2 - PURPOSE: The high percentage of patients carrying germline mutations makes pheochromocytomas/paragangliomas the most heritable of all tumors. However, there are still cases unexplained by mutations in the known genes. We aimed to identify the genetic cause of disease in patients strongly suspected of having hereditary tumors.METHODS: Whole-exome sequencing was applied to the germlines of a parent-proband trio. Genome-wide methylome analysis, RNA-seq, CRISPR/Cas9 gene editing, and targeted sequencing were also performed.RESULTS: We identified a novel de novo germline mutation in DNMT3A, affecting a highly conserved residue located close to the aromatic cage that binds to trimethylated histone H3. DNMT3A-mutated tumors exhibited significant hypermethylation of homeobox-containing genes, suggesting an activating role of the mutation. CRISPR/Cas9-mediated knock-in in HeLa cells led to global changes in methylation, providing evidence of the DNMT3A-altered function. Targeted sequencing revealed subclonal somatic mutations in six additional paragangliomas. Finally, a second germline DNMT3A mutation, also causing global tumor DNA hypermethylation, was found in a patient with a family history of pheochromocytoma.CONCLUSION: Our findings suggest that DNMT3A may be a susceptibility gene for paragangliomas and, if confirmed in future studies, would represent the first example of gain-of-function mutations affecting a DNA methyltransferase gene involved in cancer predisposition.
AB - PURPOSE: The high percentage of patients carrying germline mutations makes pheochromocytomas/paragangliomas the most heritable of all tumors. However, there are still cases unexplained by mutations in the known genes. We aimed to identify the genetic cause of disease in patients strongly suspected of having hereditary tumors.METHODS: Whole-exome sequencing was applied to the germlines of a parent-proband trio. Genome-wide methylome analysis, RNA-seq, CRISPR/Cas9 gene editing, and targeted sequencing were also performed.RESULTS: We identified a novel de novo germline mutation in DNMT3A, affecting a highly conserved residue located close to the aromatic cage that binds to trimethylated histone H3. DNMT3A-mutated tumors exhibited significant hypermethylation of homeobox-containing genes, suggesting an activating role of the mutation. CRISPR/Cas9-mediated knock-in in HeLa cells led to global changes in methylation, providing evidence of the DNMT3A-altered function. Targeted sequencing revealed subclonal somatic mutations in six additional paragangliomas. Finally, a second germline DNMT3A mutation, also causing global tumor DNA hypermethylation, was found in a patient with a family history of pheochromocytoma.CONCLUSION: Our findings suggest that DNMT3A may be a susceptibility gene for paragangliomas and, if confirmed in future studies, would represent the first example of gain-of-function mutations affecting a DNA methyltransferase gene involved in cancer predisposition.
U2 - 10.1038/s41436-018-0003-y
DO - 10.1038/s41436-018-0003-y
M3 - Article
C2 - 29740169
JO - Genetics in Medicine
JF - Genetics in Medicine
SN - 1098-3600
ER -