Abstract 1114: Polymerase epsilon (POLE) mutations and mutator phenotypes in colorectal and endometrial tumors.
Authors: Shinbrot Eve , Weinhold Nils , Schultz Nikolaus , Donehower Lawrence A. , Drummond Jennifer , Chang Kyle , Gibbs Richard , Sander Chris , Wheeler David A. 
Published: August 2014

Abstract
Colorectal rectal and endometrial cancers are divided into microsatellite instable (MSI) and microsatellite stable (MSS) types. MSS tumors have chromosome instability with mutation rates of 1-10/Mb. MSI patients have a better prognosis than MSS patients and are treated less aggressively. These tumors exhibit microsatellite instability (MSI), and the CpG island methylator phenotype (CIMP), an increased mutation rate (10-100/Mb, hypermutated). Here we demonstrate a novel class of tumor in these two diseases exhibiting an ultramutator phenotype with mutation rates exceeding 100/ Mb, MSS and chromosome stable. The ultra-high mutation rates appear to be caused by recurrent mutations in the exonuclease domain of DNA polymerase epsilon (POLE).
Matched tumor and normal whole exomes for 512 colorectal and 248 endometrial cancers (TCGA data control center) were evaluated for mutation frequency and MSI status. Among the colorectal tumors, 70/512(14%) were hyper mutated, endometroid tumors had 65/248(26%) hypermutated tumors. Within the hypermutated tumors, we identified tumors with extremely high mutation rates (100 mutations/Mb) termed “ultramutated”. 14 (3%) colorectal and 17 (7%) endometrial were ultramutated. 100% of the colorectal and endometrial ultramutated contain an exonuclease domain mutation in POLE with three recurrently mutated positions: P286R/H/S and V411L and S459F, these mutations are found exclusively in ultramutated samples. The functional relevance of these mutations has been demonstrated by previous mutational analysis of the exonuclease domain in bacteria phage (T4) yeast,and mice. Disruption of residues in the active site leads to high error rates, a mutator phenotype and tumor formation. The defined exonuclease active site aligns closely with the recurrent mutation found in the ultramutated tumors.
The ultramuted tumors have a distinct mutation spectrum, occur in context and exhibit strand biases. POLE exonuclease domain mutation tumors have high rates of C to A and T to G mutations with low rates of C to G and T to A. Changes of C to A occur predominately in context of 3’ and 5’ flanking T bases. This mutation spectrum and context of flanking bases suggests replicative strand bias. We found that mutations on the leading strand, exhibited a preference for C to A over G to T (60:40). Suggesting transcription-coupled repair (TCR) plays a role in ultramutation.
Our results demonstrate that ultra mutated tumors are driven by recurrent mutations in the exonuclease domain of DNA polymerase epsilon. The distinct mutation spectrum, strand bias and mutation context all suggest TCR is involved in ultramutated tumors. We describe a previously unknown role of POLE in colorectal tumors as well as identification of a previously undescribed hot spot for cancer mutations.
Citation Format: Eve Shinbrot, Nils Weinhold, Nikolaus Schultz, Lawrence A. Donehower, Jennifer Drummond, Kyle Chang, Richard Gibbs, Chris Sander, David A. Wheeler. Polymerase epsilon (POLE) mutations and mutator phenotypes in colorectal and endometrial tumors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1114. doi:10.1158/1538-7445.AM2013-1114