Variant filtering and prioritization
The variants identified by whole-exome sequencing where subsequently
filtered for the selection of those variants that were: 1) shared by the
cancer-affected family members sequenced; 2) carried in heterozygosis;
3) coding (frameshift, inframe, nonsense, splicing or missense) and
affecting autosomes; 4) rare (minor allele frequency (MAF)≤0.01 in the
general population) and not present in 3 or more families; 5) predicted
to be damaging by least 4 out of 5 in silico tools for missense
variants and 2 out of 2 for inframe variants, or predicted to alter
splicing for splice region variants; and 6) absent in a healthy elderly
relative sequenced (when applicable). Finally, the filtered variants
were prioritized based on the relevance of the gene and the location of
the variant in the protein structure, allowing the selection of a list
of candidate variants.