Spatial and Temporal Heterogeneity of Panel-Based Tumor Mutational Burden in Pulmonary Adenocarcinoma: Separating Biology From Technical Artifacts.


Journal

Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer
ISSN: 1556-1380
Titre abrégé: J Thorac Oncol
Pays: United States
ID NLM: 101274235

Informations de publication

Date de publication:
11 2019
Historique:
received: 02 05 2019
revised: 25 06 2019
accepted: 05 07 2019
pubmed: 28 7 2019
medline: 26 8 2020
entrez: 27 7 2019
Statut: ppublish

Résumé

Tumor mutational burden (TMB) is an emerging biomarker used to identify patients who are more likely to benefit from immuno-oncology therapy. Aside from various unsettled technical aspects, biological variables such as tumor cell content and intratumor heterogeneity may play an important role in determining TMB. TMB estimates were determined applying the TruSight Oncology 500 targeted sequencing panel. Spatial and temporal heterogeneity was analyzed by multiregion sequencing (two to six samples) of 24 pulmonary adenocarcinomas and by sequencing a set of matched primary tumors, locoregional lymph node metastases, and distant metastases in five patients. On average, a coding region of 1.28 Mbp was covered with a mean read depth of 609x. Manual validation of the mutation-calls confirmed a good performance, but revealed noticeable misclassification during germline filtering. Different regions within a tumor showed considerable spatial TMB variance in 30% (7 of 24) of the cases (maximum difference, 14.13 mut/Mbp). Lymph node-derived TMB was significantly lower (p = 0.016). In 13 cases, distinct mutational profiles were exclusive to different regions of a tumor, leading to higher values for simulated aggregated TMB. Combined, intratumor heterogeneity and the aggregated TMB could result in divergent TMB designation in 17% of the analyzed patients. TMB variation between primary tumor and distant metastases existed but was not profound. Our data show that, in addition to technical aspects such as germline filtering, the tumor content and spatially divergent mutational profiles within a tumor are relevant factors influencing TMB estimation, revealing limitations of single-sample-based TMB estimations in a clinical context.

Sections du résumé

BACKGROUND
Tumor mutational burden (TMB) is an emerging biomarker used to identify patients who are more likely to benefit from immuno-oncology therapy. Aside from various unsettled technical aspects, biological variables such as tumor cell content and intratumor heterogeneity may play an important role in determining TMB.
METHODS
TMB estimates were determined applying the TruSight Oncology 500 targeted sequencing panel. Spatial and temporal heterogeneity was analyzed by multiregion sequencing (two to six samples) of 24 pulmonary adenocarcinomas and by sequencing a set of matched primary tumors, locoregional lymph node metastases, and distant metastases in five patients.
RESULTS
On average, a coding region of 1.28 Mbp was covered with a mean read depth of 609x. Manual validation of the mutation-calls confirmed a good performance, but revealed noticeable misclassification during germline filtering. Different regions within a tumor showed considerable spatial TMB variance in 30% (7 of 24) of the cases (maximum difference, 14.13 mut/Mbp). Lymph node-derived TMB was significantly lower (p = 0.016). In 13 cases, distinct mutational profiles were exclusive to different regions of a tumor, leading to higher values for simulated aggregated TMB. Combined, intratumor heterogeneity and the aggregated TMB could result in divergent TMB designation in 17% of the analyzed patients. TMB variation between primary tumor and distant metastases existed but was not profound.
CONCLUSIONS
Our data show that, in addition to technical aspects such as germline filtering, the tumor content and spatially divergent mutational profiles within a tumor are relevant factors influencing TMB estimation, revealing limitations of single-sample-based TMB estimations in a clinical context.

Identifiants

pubmed: 31349062
pii: S1556-0864(19)30563-5
doi: 10.1016/j.jtho.2019.07.006
pii:
doi:

Substances chimiques

Biomarkers, Tumor 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

1935-1947

Informations de copyright

Copyright © 2019 International Association for the Study of Lung Cancer. Published by Elsevier Inc. All rights reserved.

Auteurs

Daniel Kazdal (D)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany; Translational Lung Research Center (TLRC) Heidelberg, German Center for Lung Research (DZL), Heidelberg, Germany.

Volker Endris (V)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Michael Allgäuer (M)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Mark Kriegsmann (M)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Jonas Leichsenring (J)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Anna-Lena Volckmar (AL)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Alexander Harms (A)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany; Translational Lung Research Center (TLRC) Heidelberg, German Center for Lung Research (DZL), Heidelberg, Germany.

Martina Kirchner (M)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Katharina Kriegsmann (K)

Department of Hematology, Oncology and Rheumatology, University Hospital Heidelberg, Heidelberg, Germany.

Olaf Neumann (O)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Regine Brandt (R)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Suranand B Talla (SB)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Eugen Rempel (E)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Carolin Ploeger (C)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Moritz von Winterfeld (M)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Petros Christopoulos (P)

Translational Lung Research Center (TLRC) Heidelberg, German Center for Lung Research (DZL), Heidelberg, Germany; Department of Thoracic Oncology, Thoraxklinik at the University Hospital Heidelberg, Heidelberg, Germany.

Diana M Merino (DM)

Friends of Cancer Research, Washington, DC.

Mark Stewart (M)

Friends of Cancer Research, Washington, DC.

Jeff Allen (J)

Friends of Cancer Research, Washington, DC.

Helge Bischoff (H)

Translational Lung Research Center (TLRC) Heidelberg, German Center for Lung Research (DZL), Heidelberg, Germany; Department of Thoracic Oncology, Thoraxklinik at the University Hospital Heidelberg, Heidelberg, Germany.

Michael Meister (M)

Translational Lung Research Center (TLRC) Heidelberg, German Center for Lung Research (DZL), Heidelberg, Germany; Translational Research Unit, Thoraxklinik at University Hospital Heidelberg, Heidelberg, Germany.

Thomas Muley (T)

Translational Research Unit, Thoraxklinik at University Hospital Heidelberg, Heidelberg, Germany.

Felix Herth (F)

Translational Lung Research Center (TLRC) Heidelberg, German Center for Lung Research (DZL), Heidelberg, Germany; Department of Pneumonology and Critical Care Medicine, Thoraxklinik at the University Hospital Heidelberg, Germany.

Roland Penzel (R)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Arne Warth (A)

Institute of Pathology, Cytopathology, and Molecular Pathology UEGP MVZ Giessen/ Wetzlar/Limburg, Germany.

Hauke Winter (H)

Translational Lung Research Center (TLRC) Heidelberg, German Center for Lung Research (DZL), Heidelberg, Germany; Department of Thoracic Surgery, Thoraxklinik at the University Hospital Heidelberg, Heidelberg, Germany.

Stefan Fröhling (S)

Department of Translational Medical Oncology, National Center for Tumor Diseases (NCT), Heidelberg, Germany; German Cancer Consortium (DKTK), Partner Site, Heidelberg, Germany.

Solange Peters (S)

Department of Oncology, Centre Hospitalier Universitaire Vaudois (CHUV) and Lausanne University, Lausanne, Switzerland.

Charles Swanton (C)

Cancer Evolution and Genome Instability Translational Cancer Therapeutics Laboratory, Francis Crick Institute, London, United Kingdom.

Michael Thomas (M)

Translational Lung Research Center (TLRC) Heidelberg, German Center for Lung Research (DZL), Heidelberg, Germany; Department of Thoracic Oncology, Thoraxklinik at the University Hospital Heidelberg, Heidelberg, Germany.

Peter Schirmacher (P)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany; German Cancer Consortium (DKTK), Partner Site, Heidelberg, Germany.

Jan Budczies (J)

Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany.

Albrecht Stenzinger (A)

Translational Lung Research Center (TLRC) Heidelberg, German Center for Lung Research (DZL), Heidelberg, Germany; German Cancer Consortium (DKTK), Partner Site, Heidelberg, Germany. Electronic address: albrecht.stenzinger@med.uni-heidelberg.de.

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Classifications MeSH