Implementing amplicon-based next generation sequencing in the diagnosis of small cell lung carcinoma metastases

Lydia Meder; Katharina König; Jana Fassunke; Luka Ozretić; Jürgen Wolf; Sabine Merkelbach-Bruse; Lukas C Heukamp; Reinhard Buettner

doi:10.1016/j.yexmp.2015.11.002

Implementing amplicon-based next generation sequencing in the diagnosis of small cell lung carcinoma metastases

Exp Mol Pathol. 2015 Dec;99(3):682-6. doi: 10.1016/j.yexmp.2015.11.002. Epub 2015 Nov 10.

Authors

Lydia Meder¹, Katharina König², Jana Fassunke¹, Luka Ozretić¹, Jürgen Wolf³, Sabine Merkelbach-Bruse¹, Lukas C Heukamp¹, Reinhard Buettner⁴

Affiliations

¹ Institute of Pathology, University Hospital Cologne, Kerpener Straße 62, 50937 Cologne, Germany; Center for Integrated Oncology Cologne/Bonn, University of Cologne, Kerpener Straße 62, 50937 Cologne, Germany; Lung Cancer Group Cologne, University of Cologne, Kerpener Straße 62, 50937 Cologne, Germany.
² Institute of Pathology, University Hospital Cologne, Kerpener Straße 62, 50937 Cologne, Germany; Center for Integrated Oncology Cologne/Bonn, University of Cologne, Kerpener Straße 62, 50937 Cologne, Germany; Lung Cancer Group Cologne, University of Cologne, Kerpener Straße 62, 50937 Cologne, Germany; Labor Dr. Quade und Kollegen GmbH, Aachener Straße 338, 50933 Cologne, Germany.
³ Center for Integrated Oncology Cologne/Bonn, University of Cologne, Kerpener Straße 62, 50937 Cologne, Germany; Lung Cancer Group Cologne, University of Cologne, Kerpener Straße 62, 50937 Cologne, Germany; Clinic for Internal Medicine I, University of Cologne, Kerpener Straße 62, 50937 Cologne, Germany.
⁴ Institute of Pathology, University Hospital Cologne, Kerpener Straße 62, 50937 Cologne, Germany; Center for Integrated Oncology Cologne/Bonn, University of Cologne, Kerpener Straße 62, 50937 Cologne, Germany; Lung Cancer Group Cologne, University of Cologne, Kerpener Straße 62, 50937 Cologne, Germany. Electronic address: Reinhard.Buettner@uk-koeln.de.

PMID: 26546837
DOI: 10.1016/j.yexmp.2015.11.002

Abstract

Small cell lung carcinoma (SCLC) is the most aggressive entity of lung cancer. Rapid cancer progression and early formation of systemic metastases drive the deadly outcome of SCLC. Recent advances in identifying oncogenes by cancer whole genome sequencing improved the understanding of SCLC carcinogenesis. However, tumor material is often limited in the clinic. Thus, it is a compulsive issue to improve SCLC diagnostics by combining established immunohistochemistry and next generation sequencing. We implemented amplicon-based next generation deep sequencing in our routine diagnostics pipeline to analyze RB1, TP53, EP300 and CREBBP, frequently mutated in SCLC. Thereby, our pipeline combined routine SCLC histology and identification of somatic mutations. We comprehensively analyzed fifty randomly collected SCLC metastases isolated from trachea and lymph nodes in comparison to specimens derived from primary SCLC. SCLC lymph node metastases showed enhanced proliferation and frequently a collapsed keratin cytoskeleton compared to SCLC metastases isolated from trachea. We identified characteristic synchronous mutations in RB1 and TP53 and non-synchronous CREBBP and EP300 mutations. Our data showed the benefit of implementing deep sequencing into routine diagnostics. We here identify oncogenic drivers and simultaneously gain further insights into SCLC tumor biology.

Keywords: Amplicon-based next generation sequencing; Metastasis; Retinoblastoma protein; Routine diagnostics; Small cell lung cancer; p53.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA Mutational Analysis / methods*
High-Throughput Nucleotide Sequencing / methods*
Humans
Lung Neoplasms / genetics*
Lung Neoplasms / pathology
Neoplasm Metastasis / diagnosis
Neoplasm Metastasis / genetics*
Small Cell Lung Carcinoma / genetics*
Small Cell Lung Carcinoma / pathology