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Molecular subtype switching in early-stage gastric cancers with multiple occurrences

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Abstract

Background

Multiple gastric cancers at the same time (synchronous) or recurrence after 1 year (metachronous) are frequently encountered. Since their genetic profiles were not well elucidated, we molecularly subtyped the genetic events of synchronous and metachronous early-stage gastric cancers.

Methods

We studied mismatch repair (MMR) genes in 84 tumors from 31 patients (15 synchronous and 16 metachronous) by immunohistochemistry. We performed microsatellite instability analysis and targeted sequencing of 58 significantly mutated genes (SMGs) in 35 tumors from thirteen patients. Genomic data from TCGA were used for comparisons with advanced-stage cancers.

Results

Among the 31 patients, at least one deficient-MMR (dMMR) tumor was observed in eight (26%). Of eight patients, seven showed a mixture of proficient-MMR (pMMR) and dMMR tumors. The one case with only dMMR had six recurrent tumors within 2 years. To further subtype, we sequenced 58 SMGs in 35 samples (25 pMMR and 10 dMMR) from thirteen patients. In 35 samples, 163 mutations were identified, but none matched in almost cases, strongly indicating different clonal origins, whether synchronous or metachronous occurrences. Of the 25 pMMR cases, 1 belonged to Epstein–Barr virus (EBV), 24 belonged to chromosomal instability (CIN) subtypes. Of the thirteen cases, repetitive CIN, a mixture of CIN and MSI, a mixture of CIN and EBV, and repetitive MSI were observed in nine (70%), two (15%), one (8%) and one (8%), respectively.

Conclusions

Despite multiple tumors occurring in the same patient simultaneously or several years apart, clonal origin was totally different. ‘Switching’ or ‘mixing’ of dMMR and pMMR, EBV or CIN occurred, which had clinical relevance with regard to immunotherapy.

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Abbreviations

MMR:

Mismatch repair

SMGs:

Significantly mutated genes

dMMR:

Deficient-MMR

pMMR:

Proficient-MMR

EBV:

Epstein–Barr virus

EBER:

Epstein–Barr virus encoded small mRNA’s

MSI:

Microsatellite instability

CIN:

Chromosomal instability

ESD:

Endoscopic submucosal dissection

TCGA:

The Cancer Genome Atlas

ICGC:

International Cancer Genome Consortium

IHC:

Immunohistochemistry

FFPE:

Formalin-fixed paraffin-embedded

COSMIC:

Catalog of Somatic Mutations in Cancer

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Acknowledgements

This study was supported by a Grant-in-Aid for Genome Research Project from Yamanashi Prefecture (to M.O. and Y.H.), The Japan Society for the Promotion of Science (JSPS) KAKENHI Early-Career Scientists (Grant Number JP18K16292 to Y.H.), Research Grant for Young Scholars (to Y.H.), The YASUDA Medical Foundation (to Y.H.) and The Uehara Memorial Foundation (to Y.H.). We thank all medical and ancillary hospital staff and the patients for consenting to participate. We also thank Kanoko Sano, Saki Hiraga, Shunsuke Watanabe, and Eri Ishii for technical support. Finally, we thank James P. Mahaffey, Ph.D., from Edanz Group (www.edanzediting.com/ac) for editing a draft of this manuscript.

Author information

Author notes

  1. Shinya Takaoka and Yosuke Hirotsu have contributed equally to this study.

Authors and Affiliations

  1. Department of Gastroenterology, Yamanashi Central Hospital, 1-1-1 Fujimi, Kofu, Yamanashi, Japan

    Shinya Takaoka, Hiroshi Ohyama, Hitoshi Mochizuki, Hiroshi Ashizawa, Dai Yoshimura, Keiko Nakagomi, Kenji Hosoda, Yoji Suzuki, Yuichiro Kojima & Masao Omata

  2. Genome Analysis Center, Yamanashi Central Hospital, 1-1-1 Fujimi, Kofu, Yamanashi, Japan

    Yosuke Hirotsu, Hitoshi Mochizuki & Kenji Amemiya

  3. Department of Gastroenterology, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba, Japan

    Hiroshi Ohyama

  4. Department of Pathology, Yamanashi Central Hospital, 1-1-1 Fujimi, Kofu, Yamanashi, Japan

    Toshio Oyama

  5. The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan

    Masao Omata

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  1. Shinya Takaoka
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Corresponding author

Correspondence to Yosuke Hirotsu.

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Supplementary material 1 (XLSX 18 kb)

535_2019_1547_MOESM2_ESM.pdf

Supplemental Figure 1. Relationship between mutated genes and variant allele fractions. Average of variant allele fractions and the number of mutations were plotted (PDF 9 kb)

535_2019_1547_MOESM3_ESM.pdf

Epstein–Barr virus (EBV) infection status was determined by in situ hybridization targeting EBV encoded small mRNA’s (EBER). Representative images for hematoxylin-eosin staining and EBER in situ hybridization performed in EBV-positive (T1 in Case #3) and EBV-negative (T1 in Case #16) gastric cancer tissues. Scale bar: 100 μm (PDF 205 kb)

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Takaoka, S., Hirotsu, Y., Ohyama, H. et al. Molecular subtype switching in early-stage gastric cancers with multiple occurrences. J Gastroenterol 54, 674–686 (2019). https://doi.org/10.1007/s00535-019-01547-z

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  • DOI: https://doi.org/10.1007/s00535-019-01547-z

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