title: Implementing amplicon-based next generation sequencing in the diagnosis of small cell lung carcinoma metastases.
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.
authors.affiliation: 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.
authors.forename: Lydia
authors.initials: L
authors.lastname: Meder
authors.affiliation: 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.
authors.forename: Katharina
authors.initials: K
authors.lastname: König
authors.affiliation: 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.
authors.forename: Jana
authors.initials: J
authors.lastname: Fassunke
authors.affiliation: 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.
authors.forename: Luka
authors.initials: L
authors.lastname: Ozretić
authors.affiliation: 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.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Wolf
authors.affiliation: 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.
authors.forename: Sabine
authors.initials: S
authors.lastname: Merkelbach-Bruse
authors.affiliation: 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.
authors.forename: Lukas C
authors.initials: LC
authors.lastname: Heukamp
authors.affiliation: 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.
authors.forename: Reinhard
authors.initials: R
authors.lastname: Buettner
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology; D009362:Neoplasm Metastasis / Q000175:diagnosis / Q000235:genetics*; D055752:Small Cell Lung Carcinoma / Q000235:genetics* / Q000473:pathology
keywords: Amplicon-based next generation sequencing; Metastasis; Retinoblastoma protein; Routine diagnostics; Small cell lung cancer; p53