Whole exome sequencing of ENU-induced thrombosis modifier mutations in the mouse

摘要: Although the Factor V Leiden (FVL) gene variant is most prevalent genetic risk factor for venous thrombosis, only 10% of FVL carriers will experience such an event in their lifetime. To identify potential modifier genes contributing to this incomplete penetrance, we took advantage a perinatal synthetic lethal thrombosis phenotype mice homozygous (F5L/L) and haploinsufficient tissue pathway inhibitor (Tfpi+/-) perform sensitized dominant ENU mutagenesis screen. Linkage analysis conducted 3 largest pedigrees generated from surviving F5L/L Tfpi+/- ("rescues") using ENU-induced coding variants as markers was unsuccessful identifying major suppressor loci. Whole exome sequencing applied DNA 107 rescue candidate enriched mutations. A total 3,481 potentially deleterious were identified 2,984 genes. After correcting size multiple testing, Arl6ip5 gene, though not reaching genome-wide significance. Evaluation CRISPR/Cas9 induced loss function top 6 failed demonstrate clear phenotype. However, maternally inherited (not ENU-induced) de novo mutation (Plcb4R335Q) exhibited significant co-segregation with (p=0.003) corresponding pedigree. Thrombosis suppression by heterozygous Plcb4 confirmed through independent, CRISPR/Cas9-induced (p=0.01).