PT  - JOURNAL ARTICLE
AU  - Kärt Tomberg
AU  - Randal J. Westrick
AU  - Emilee N. Kotnik
AU  - David R. Siemieniak
AU  - Guojing Zhu
AU  - Thomas L. Saunders
AU  - Audrey C. Cleuren
AU  - David Ginsburg
TI  - Whole exome sequencing of ENU-induced thrombosis modifier mutations in the mouse
AID  - 10.1101/174086
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 174086
4099  - http://biorxiv.org/content/early/2017/08/09/174086.short
4100  - http://biorxiv.org/content/early/2017/08/09/174086.full
AB  - Although the Factor V Leiden (FVL) gene variant is the most common genetic risk factor for venous thrombosis, only 10% of these individuals will experience such an event in their lifetime. In order to identify potential FVL modifier genes contributing to this incomplete penetrance, we performed a sensitized dominant ENU mutagenesis screen, based on the perinatal synthetic lethal thrombosis previously observed in mice homozygous for FVL (F5L/L) and haploinsufficient for tissue factor pathway inhibitor (Tfpi+/-). Whole exome sequencing was applied to DNA from 107 viable F5L/L Tfpi+/- progeny (‘rescues’) to identify candidate genes that are enriched for ENU mutations. A total of 3481 potentially deleterious candidate ENU variants were identified in 2984 genes. After correcting for gene size and multiple testing, Arl6ip5 was identified as the most enriched gene but did not meet genome-wide significance and validation of the top 6 genes using independent CRISPR/Cas9 induced mutations failed to demonstrate clear rescue phenotype. Linkage analysis conducted in 3 largest pedigrees generated from the rescue mice using ENU-induced coding variants as genetic markers did not map the corresponding suppressor loci. However, in one of the pedigrees, a maternally inherited (not ENU-induced) de novo mutation (Plcb4R335Q) exhibited significant co-segregation with the rescue phenotype (p=0.003).