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. 2024 Apr 17;24(1):489.
doi: 10.1186/s12885-024-12250-5.

Utility of bronchoscopically obtained frozen cytology pellets for next-generation sequencing

Chihiro Mimura  1 , Rei Takamiya  1 , Shodai Fujimoto  1 , Takafumi Fukui  1 , Atsuhiko Yatani  1 , Jun Yamada  1 , Mizuki Takayasu  1 , Naoya Takata  1 , Hiroki Sato  1 , Kiyoko Fukuda  1 , Koichi Furukawa  1 , Daisuke Hazama  1 , Naoko Katsurada  1 , Masatsugu Yamamoto  1 , Shingo Matsumoto  2 , Koichi Goto  2 , Motoko Tachihara  3
Affiliations

Utility of bronchoscopically obtained frozen cytology pellets for next-generation sequencing

Chihiro Mimura et al. BMC Cancer. .

Abstract

Background: Next-generation sequencing (NGS) is essential for lung cancer treatment. It is important to collect sufficient tissue specimens, but sometimes we cannot obtain large enough samples for NGS analysis. We investigated the yield of NGS analysis by frozen cytology pellets using an Oncomine Comprehensive Assay or Oncomine Precision Assay.

Methods: We retrospectively enrolled patients with lung cancer who underwent bronchoscopy at Kobe University Hospital and were enrolled in the Lung Cancer Genomic Screening Project for Individualized Medicine. We investigated the amount of extracted DNA and RNA and determined the NGS success rates. We also compared the amount of DNA and RNA by bronchoscopy methods. To create the frozen cytology pellets, we first effectively collected the cells and then quickly centrifuged and cryopreserved them.

Results: A total of 132 patients were enrolled in this study between May 2016 and December 2022; of them, 75 were subjected to frozen cytology pellet examinations and 57 were subjected to frozen tissue examinations. The amount of DNA and RNA obtained by frozen cytology pellets was nearly equivalent to frozen tissues. Frozen cytology pellets collected by endobronchial ultrasound-guided transbronchial needle aspiration yielded significantly more DNA than those collected by transbronchial biopsy methods. (P < 0.01) In RNA content, cytology pellets were not inferior to frozen tissue. The success rate of NGS analysis with frozen cytology pellet specimens was comparable to the success rate of NGS analysis with frozen tissue specimens.

Conclusions: Our study showed that frozen cytology pellets may have equivalent diagnostic value to frozen tissue for NGS analyses. Bronchial cytology specimens are usually used only for cytology, but NGS analysis is possible if enough cells are collected to create pellet specimens. In particular, the frozen cytology pellets obtained by endobronchial ultrasound-guided transbronchial needle aspiration yielded sufficient amounts of DNA.

Trial registration: This was registered with the University Medical Hospital Information Network in Japan (UMINCTR registration no. UMIN000052050).

Keywords: Cytology; Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA); Lung cancer; Next-generation sequencing (NGS); Pellet specimen.

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Conflict of interest statement

MT reports grants from AstraZeneca KK, Chugai Pharmaceutical Co., Ltd., Eli Lilly Japan Co., Ltd., honoraria for lecture from Eli Lilly Japan Co., Ltd., Ono Pharmaceutical Co., Ltd., Bristol-Myers Squibb Co., Ltd., Chugai Pharmaceutical Co., Ltd., AstraZeneca KK, MSD KK, Novartis Pharmaceuticals K.K, Takeda Pharmaceutical Co., Ltd., Taiho Pharmaceutical Co., Ltd., Nippon Boehringer Ingelheim Co.,Ltd., Daiichi Sankyo, Pfizer Japan Inc., and Janssen Pharmaceutical K.K. SM reports grants from Chugai Pharma, Janssen Pharmaceutical, MSD, Novartis, honoraria for lecture from Eli Lilly, Merck Biopharma, Chugai Pharma, Novartis pharma, Guardant Health, and AstraZeneca. KG reports all support for the present manuscript from Thermo Fisher Scientific K.K., grants from Amgen Inc., Amgen K.K., Amgen Astellas BioPharma K.K., AstraZeneca K.K., Bayer Yakuhin, Ltd., Boehringer Ingelheim Japan, Inc., Bristol-Myers Squibb K.K., Chugai Pharmaceutical Co., Ltd., Craif Inc., DAIICHI SANKYO Co., Ltd., Eisai Co., Ltd., Eli Lilly Japan K.K., Haihe Biopharma Co., Ltd., Ignyta,Inc., Janssen Pharmaceutical K.K., KISSEI PHARMACEUTICAL CO., LTD., Kyowa Kirin Co., Ltd., Life Technologies Japan Ltd., Loxo Oncology, Inc., LSI Medience Corporation., MEDICAL& BIOLOGICAL LABORATORIES CO., LTD., Merck Biopharma Co., Ltd., Merus N.V., MSD K.K., Novartis Pharma K.K., Ono Pharmaceutical Co., Ltd., Pfizer Japan Inc., Pfizer R&D Japan G.K., Precision Medicine Asia Co., Ltd., RIKEN GENESIS CO., LTD., Sumitomo Pharma Co., Ltd., Spectrum Pharmaceuticals, Inc., Sysmex Corporation., Taiho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Co., Ltd., Turning Point Therapeutics,Inc., Xcoo, Inc., honoraria for lecture from Amgen K.K., Amoy Diagnosties Co.,Ltd., AstraZeneca K.K., Bayer U.S., Boehringer Ingelheim Japan, Inc., Bristol-Myers Squibb K.K., Chugai Pharmaceutical Co., Ltd., DAIICHI SANKYO Co., Ltd., Eisai Co., Ltd., Eli Lilly Japan K.K., Guardant Health Inc., Janssen Pharmaceutical K.K., Merck Biopharma Co., Ltd., Nippon Kayaku Co.,Ltd., Novartis Pharma K.K., Ono Pharmaceutical Co., Ltd., Otsuka Pharmaceutical Co., Ltd., RIKEN GENESIS CO., LTD., Taiho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Co., Ltd., Thermo Fisher Scientific K.K., participation on a Data Safety Monitoring Board or Advisory Board from Amgen Inc., Amgen K.K., DAIICHI SANKYO Co., Ltd., Eli Lilly Japan K.K., Haihe Biopharma Co., Ltd., Janssen Pharmaceutical K.K., SYNEOS HEALTH CLINICAL K.K. Other authors declare no conflicts of interest.

Figures

Fig. 1
Fig. 1
How to make the pellets. Legends: Schema of the procedure used to make the frozen cytology pellets. First, we smeared the cells on the glass slide. Next, we washed the devices in saline on ice. We collected the remaining cells in saline, and cryopreserved them as cytology pellets
Fig. 2
Fig. 2
Extracted DNA and RNA yielded according to the procedure. Legends: TBB methods were further divided into TBB (ultrathin), EBUS-GS-TBB (thin), EBUS-GS-TBB (thick), EBB, and TBB under X-ray fluoroscopy. EBB, endobronchial biopsy; EBUS-GS-TBB, endobronchial ultrasound with a guide sheath transbronchial biopsy; TBB, transbronchial biopsy
Fig. 3
Fig. 3
Genetic alterations obtained by NGS analysis. Legends: The percentages and the number of patients of targetable genetic alterations in non-squamous non-small cell lung cancer (N = 80). Others included two driver mutations: GNAS and PIKC3A
See this image and copyright information in PMC

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