A Functional Assay-based Procedure to Classify Mismatch Repair Gene Variants in Lynch Syndrome

Overview

Journal Genet Med

Publisher Elsevier

Specialty Genetics

Date 2018 Dec 4

PMID 30504929

Citations 24

Authors

Mark Drost

Yvonne Tiersma

Bryony A Thompson

Jane H Frederiksen

Guido Keijzers

Dylan Glubb

Scott Kathe

Jan Osinga

Helga Westers

Lisa Pappas

Kenneth M Boucher

Siska Molenkamp

Jose B Zonneveld

Christi J van Asperen

David E Goldgar

Susan S Wallace

Rolf H Sijmons

Amanda B Spurdle

Lene J Rasmussen

Marc S Greenblatt

Niels de Wind

Sean V Tavtigian

Affiliations

Soon will be listed here.

Abstract

Purpose: To enhance classification of variants of uncertain significance (VUS) in the DNA mismatch repair (MMR) genes in the cancer predisposition Lynch syndrome, we developed the cell-free in vitro MMR activity (CIMRA) assay. Here, we calibrate and validate the assay, enabling its integration with in silico and clinical data.

Methods: Two sets of previously classified MLH1 and MSH2 variants were selected from a curated MMR gene database, and their biochemical activity determined by the CIMRA assay. The assay was calibrated by regression analysis followed by symmetric cross-validation and Bayesian integration with in silico predictions of pathogenicity. CIMRA assay reproducibility was assessed in four laboratories.

Results: Concordance between the training runs met our prespecified validation criterion. The CIMRA assay alone correctly classified 65% of variants, with only 3% discordant classification. Bayesian integration with in silico predictions of pathogenicity increased the proportion of correctly classified variants to 87%, without changing the discordance rate. Interlaboratory results were highly reproducible.

Conclusion: The CIMRA assay accurately predicts pathogenic and benign MMR gene variants. Quantitative combination of assay results with in silico analysis correctly classified the majority of variants. Using this calibration, CIMRA assay results can be integrated into the diagnostic algorithm for MMR gene variants.

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Functional analysis of MMR gene VUS from potential Lynch syndrome patients.

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