Harmonization and Standardization of Panel-Based Tumor Mutational Burden Measurement: Real-World Results and Recommendations of the Quality in Pathology Study.

Abstract

INTRODUCTION:

Tumor mutational burden (TMB) is a quantitative assessment of the number of somatic mutations within a tumor genome. Immunotherapy benefit has been associated with TMB assessed by whole-exome sequencing (wesTMB) and gene panel sequencing (psTMB). The initiatives of Quality in Pathology (QuIP) and Friends of Cancer Research have jointly addressed the need for harmonization among TMB testing options in tissues. This QuIP study identifies critical sources of variation in psTMB assessment.

METHODS:

A total of 20 samples from three tumor types (lung adenocarcinoma, head and neck squamous cell carcinoma, and colon adenocarcinoma) with available WES data were analyzed for psTMB using six panels across 15 testing centers. Interlaboratory and interplatform variation, including agreement on variant calling and TMB classification, were investigated. Bridging factors to transform psTMB to wesTMB values were empirically derived. The impact of germline filtering was evaluated.

RESULTS:

Sixteen samples had low interlaboratory and interpanel psTMB variation, with 87.7% of pairwise comparisons revealing a Spearman's ρ greater than 0.6. A wesTMB cut point of 199 missense mutations projected to psTMB cut points between 7.8 and 12.6 mutations per megabase pair; the corresponding psTMB and wesTMB classifications agreed in 74.9% of cases. For three-tier classification with cut points of 100 and 300 mutations, agreement was observed in 76.7%, weak misclassification in 21.8%, and strong misclassification in 1.5% of cases. Confounders of psTMB estimation included fixation artifacts, DNA input, sequencing depth, genome coverage, and variant allele frequency cut points.

CONCLUSIONS:

This study provides real-world evidence that all evaluated panels can be used to estimate TMB in a routine diagnostic setting and identifies important parameters for reliable tissue TMB assessment that require careful control. As complex or composite biomarkers beyond TMB are likely playing an increasing role in therapy prediction, the efforts by QuIP and Friends of Cancer Research also delineate a general framework and blueprint for the evaluation of such assays.

• Authors: Albrecht Stenzinger, Volker Endris, Jan Budczies, Sabine Merkelbach-Bruse, Daniel Kazdal, Wolfgang Dietmaier, Nicole Pfarr, Udo Siebolts, Michael Hummel, Sylvia Herold, Johanna Andreas, Martin Zoche, Lars Tögel, Eugen Rempel, Jörg Maas, Diana Merino, Mark Stewart, Karim Zaoui, Matthias Schlesner, Hanno Glimm, Stefan Fröhling, Jeff Allen, David Horst, Gustavo Baretton, Claudia Wickenhauser, Markus Tiemann, Matthias Evert, Holger Moch, Thomas Kirchner, Reinhard Büttner, Peter Schirmacher, Andreas Jung, Florian Haller, Wilko Weichert, Manfred Dietel
• Journal: Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer (J Thorac Oncol) Vol. 15 Issue 7 Pg. 1177-1189 (07 2020) ISSN: 1556-1380 [Electronic] United States
• PMID: 32119917 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2020 International Association for the Study of Lung Cancer. Published by Elsevier Inc. All rights reserved.

Chemical References

• Biomarkers, Tumor

Topics

• Biomarkers, Tumor (genetics)
• Humans
• Lung Neoplasms (genetics)
• Mutation
• Reference Standards
• Exome Sequencing