Suitability of Bronchoscopic Biopsy Tissue Samples for Next-Generation Sequencing.

Shuji Murakami,Masaki Suzuki,Ryou Usui,Terufumi Kato,Tomoyuki Yokose,Daiji Nemoto,Yoshiro Nakahara,Haruhiro Saito,Tetsuro Kondo

doi:10.3390/diagnostics11030391

Abstract

A sufficiently large tissue sample is required to perform next-generation sequencing (NGS) with a high success rate, but the majority of patients with advanced non-small-cell lung cancer (NSCLC) are diagnosed with small biopsy specimens. Biopsy samples were collected from 184 patients with bronchoscopically diagnosed NSCLC. The tissue surface area, tumor cell count, and tumor content rate of each biopsy sample were evaluated. The impact of the cut-off criteria for the tissue surface area (≥1 mm2) and tumor content rate (≥30%) on the success rate of the Oncomine Dx Target Test (ODxTT) was evaluated. The mean tissue surface area of the transbronchial biopsies was 1.23 ± 0.85 mm2 when small endobronchial ultrasonography with a guide sheath (EBUS-GS) was used, 2.16 ± 1.49 mm2 with large EBUS-GS, and 1.81 ± 0.75 mm2 with endobronchial biopsy (EBB). The proportion of samples with a tissue surface area of ≥1 mm2 was 48.8% for small EBUS-GS, 79.2% for large EBUS-GS, and 78.6% for EBB. Sixty-nine patients underwent ODxTT. The success rate of DNA sequencing was 84.1% and that of RNA sequencing was 92.7% over all patients. The success rate of DNA (RNA) sequencing was 57.1% (71.4%) for small EBUS-GS (n = 14), 93.4% (96.9%) for large EBUS-GS (n = 32), 62.5% (100%) for EBB (n = 8), and 100% (100%) for endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) (n = 15). Regardless of the device used, a tissue surface area of ≥ 1 mm2 is adequate for samples to be tested with NGS.

Highlights

(n = 43), small endobronchial ultrasonography with a guide sheath (EBUS-GS) (n = 77), endobronchial biopsy (EBB) (n = 14), or EBUS-TBNA (n = 50), and 165 patients were initially diagnosed with bronchoscopy
Our results indicate that larger tissue samples should be collected to ensure successful next-generation sequencing (NGS) analyses, the minimum tumor sample size required for Oncomine Dx Target Test (ODxTT) is still unclear
A sufficient size of a tumor sample is required for successful NGS testing

Summary

Introduction

Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Non-small-cell lung cancer (NSCLC) accounts for approximately 85% of lung cancer cases and is the leading cause of cancer-related deaths worldwide [1]. A large majority of patients with NSCLC are diagnosed at an advanced incurable stage, with a poor prognosis. Conventional cytotoxic chemotherapy has provided clinical benefit for or prolonged the survival of patients with advanced NSCLC, but the evidence for its clinical benefit is limited, with an objective response rate of 20–40% and a median overall survival (OS) of

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Conclusion