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Surface microstructures are associated with mutational intratumoral heterogeneity in colorectal tumors

Journal of Gastroenterology volume 53, pages 1241–1252 (2018)Cite this article

Abstract

Background

Recent studies revealed that colorectal tumors are composed of genetically diverse subclones. We aimed to clarify whether the surface microstructures of colorectal tumors are associated with genetic intratumoral heterogeneity (ITH).

Methods

The surface microstructures (pit patterns) of colorectal tumors were observed using magnifying endoscopy, and biopsy specimens were obtained from respective areas when tumors exhibited multiple pit patterns. A total of 711 specimens from 477 colorectal tumors were analyzed for BRAF, KRAS and TP53 mutations using pyrosequencing and direct sequencing. A panel of cancer-related genes was analyzed through targeted sequencing in 7 tumors.

Results

Colorectal tumors with multiple pit patterns exhibited more advanced pit patterns and higher frequencies of KRAS and/or TP53 mutations than tumors with a single pit pattern. In tumors with multiple pit patterns, mutations were observed as public (common to all areas) or private (specific to certain areas), and private KRAS and/or TP53 mutations were often variable and unrelated to the pit pattern grade. Notably, invasive CRCs frequently exhibited public TP53 mutations, even in adenomatous areas, which is indicative of their early malignant potential. Targeted sequencing revealed additional public and private mutations in tumors with multiple pit patterns, indicating their single clonal origin.

Conclusions

Our results suggest intratumoral pit pattern variation does not simply reflect the process of colorectal tumor evolution, but instead represents genetically diverse subclones, and this diversity may be associated with malignant potential.

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References

  1. Fearon ER, Vogelstein B. A genetic model for colorectal tumorigenesis. Cell. 1990;61:759–67.

    Article  CAS  Google Scholar 

  2. Jones S, Chen WD, Parmigiani G, et al. Comparative lesion sequencing provides insights into tumor evolution. Proc Natl Acad Sci USA. 2008;105:4283–8.

    Article  Google Scholar 

  3. Smith G, Carey FA, Beattie J, et al. Mutations in APC, Kirsten-ras, and p53–alternative genetic pathways to colorectal cancer. Proc Natl Acad Sci USA. 2002;99:9433–8.

    Article  CAS  Google Scholar 

  4. Sottoriva A, Kang H, Ma Z, et al. A Big Bang model of human colorectal tumor growth. Nat Genet. 2015;47:209–16.

    Article  CAS  Google Scholar 

  5. Kim TM, Jung SH, An CH, et al. Subclonal genomic architectures of primary and metastatic colorectal cancer based on intratumoral genetic heterogeneity. Clin Cancer Res. 2015;21:4461–72.

    Article  CAS  Google Scholar 

  6. Uchi R, Takahashi Y, Niida A, et al. Integrated multiregional analysis proposing a new model of colorectal cancer evolution. PLoS Genet. 2016;12:e1005778.

    Article  CAS  Google Scholar 

  7. Yachida S, Jones S, Bozic I, et al. Distant metastasis occurs late during the genetic evolution of pancreatic cancer. Nature. 2010;467:1114–7.

    Article  CAS  Google Scholar 

  8. Gerlinger M, Rowan AJ, Horswell S, et al. Intratumor heterogeneity and branched evolution revealed by multiregion sequencing. N Engl J Med. 2012;366:883–92.

    Article  CAS  Google Scholar 

  9. Hao JJ, Lin DC, Dinh HQ, et al. Spatial intratumoral heterogeneity and temporal clonal evolution in esophageal squamous cell carcinoma. Nat Genet. 2016;48:1500–7.

    Article  CAS  Google Scholar 

  10. Sievers CK, Leystra AA, Clipson L, et al. Understanding intratumoral heterogeneity: lessons from the Analysis of at-risk tissue and premalignant lesions in the colon. Cancer Prev Res (Phila). 2016;9:638–41.

    Article  CAS  Google Scholar 

  11. Ciardiello F, Tortora G. EGFR antagonists in cancer treatment. N Engl J Med. 2008;358:1160–74.

    Article  CAS  Google Scholar 

  12. Karapetis CS, Khambata-Ford S, Jonker DJ, et al. K-ras mutations and benefit from cetuximab in advanced colorectal cancer. N Engl J Med. 2008;359:1757–65.

    Article  CAS  Google Scholar 

  13. Turner NC, Reis-Filho JS. Genetic heterogeneity and cancer drug resistance. Lancet Oncol. 2012;13:e178–85.

    Article  Google Scholar 

  14. Baldus SE, Schaefer KL, Engers R, et al. Prevalence and heterogeneity of KRAS, BRAF, and PIK3CA mutations in primary colorectal adenocarcinomas and their corresponding metastases. Clin Cancer Res. 2010;16:790–9.

    Article  CAS  Google Scholar 

  15. Mancuso A, Sollami R, Recine F, et al. Patient with colorectal cancer with heterogeneous KRAS molecular status responding to cetuximab-based chemotherapy. J Clin Oncol. 2010;28:e756–8.

    Article  Google Scholar 

  16. Artale S, Sartore-Bianchi A, Veronese SM, et al. Mutations of KRAS and BRAF in primary and matched metastatic sites of colorectal cancer. J Clin Oncol. 2008;26:4217–9.

    Article  Google Scholar 

  17. Thirlwell C, Will OC, Domingo E, et al. Clonality assessment and clonal ordering of individual neoplastic crypts shows polyclonality of colorectal adenomas. Gastroenterology. 2010;138:1441-54, 54 e1-7.

    Article  CAS  Google Scholar 

  18. Kang H, Salomon MP, Sottoriva A, et al. Many private mutations originate from the first few divisions of a human colorectal adenoma. J Pathol. 2015;237:355–62.

    Article  CAS  Google Scholar 

  19. Sievers CK, Zou LS, Pickhardt PJ, et al. Subclonal diversity arises early even in small colorectal tumours and contributes to differential growth fates. Gut. 2016. https://doi.org/10.1136/gutjnl-2016-312232.

    Article  PubMed  PubMed Central  Google Scholar 

  20. Kim TM, An CH, Rhee JK, et al. Clonal origins and parallel evolution of regionally synchronous colorectal adenoma and carcinoma. Oncotarget. 2015;6:27725–35.

    PubMed  PubMed Central  Google Scholar 

  21. Kudo S, Hirota S, Nakajima T, et al. Colorectal tumours and pit pattern. J Clin Pathol. 1994;47:880–5.

    Article  CAS  Google Scholar 

  22. Kudo S, Lambert R, Allen JI, et al. Nonpolypoid neoplastic lesions of the colorectal mucosa. Gastrointest Endosc. 2008;68:S3–47.

    Article  Google Scholar 

  23. Matsuda T, Fujii T, Saito Y, et al. Efficacy of the invasive/non-invasive pattern by magnifying chromoendoscopy to estimate the depth of invasion of early colorectal neoplasms. Am J Gastroenterol. 2008;103:2700–6.

    Article  Google Scholar 

  24. Kimura T, Yamamoto E, Yamano HO, et al. A novel pit pattern identifies the precursor of colorectal cancer derived from sessile serrated adenoma. Am J Gastroenterol. 2012;107:460–9.

    Article  CAS  Google Scholar 

  25. Yamamoto E, Suzuki H, Yamano HO, et al. Molecular dissection of premalignant colorectal lesions reveals early onset of the CpG island methylator phenotype. Am J Pathol. 2012;181:1847–61.

    Article  CAS  Google Scholar 

  26. Kamimae S, Yamamoto E, Kai M, et al. Epigenetic silencing of NTSR1 is associated with lateral and noninvasive growth of colorectal tumors. Oncotarget. 2015;6:29975–90.

    Article  Google Scholar 

  27. Tanaka Y, Yamano HO, Yamamoto E, et al. Endoscopic and molecular characterization of colorectal sessile serrated adenoma/polyps with cytological dysplasia. Gastrointest Endosc. 2017;86:1131–8.

    Article  Google Scholar 

  28. Endoscopic Classification Review. G. Update on the paris classification of superficial neoplastic lesions in the digestive tract. Endoscopy. 2005;37:570–8.

    Article  Google Scholar 

  29. Kudo S, Tamura S, Nakajima T, et al. Diagnosis of colorectal tumorous lesions by magnifying endoscopy. Gastrointest Endosc. 1996;44:8–14.

    Article  CAS  Google Scholar 

  30. Aoki H, Yamamoto E, Yamano HO, et al. Subtypes of the Type II Pit Pattern Reflect Distinct Molecular Subclasses in the Serrated Neoplastic Pathway. Dig Dis Sci. 2018. https://doi.org/10.1007/s10620-018-5016-5.

    Article  PubMed  Google Scholar 

  31. Bosman FT, World Health Organization., International Agency for Research on Cancer. WHO classification of tumours of the digestive system. 4th ed. Lyon: International Agency for Research on Cancer; 2010. 417 p. p.

  32. Cancer Genome Atlas. N. Comprehensive molecular characterization of human colon and rectal cancer. Nature. 2012;487:330–7.

    Article  CAS  Google Scholar 

  33. Novelli MR, Williamson JA, Tomlinson IP, et al. Polyclonal origin of colonic adenomas in an XO/XY patient with FAP. Science. 1996;272:1187–90.

    Article  CAS  Google Scholar 

  34. Williams MJ, Werner B, Barnes CP, et al. Identification of neutral tumor evolution across cancer types. Nat Genet. 2016;48:238–44.

    Article  CAS  Google Scholar 

  35. Cross W, Graham TA, Wright NA. New paradigms in clonal evolution: punctuated equilibrium in cancer. J Pathol. 2016;240:126–36.

    Article  Google Scholar 

  36. Je IJ, Vermeulen L, Meijer GA, et al. Serrated neoplasia-role in colorectal carcinogenesis and clinical implications. Nat Rev Gastroenterol Hepatol. 2015;12:401–9.

    Article  CAS  Google Scholar 

  37. Thliveris AT, Halberg RB, Clipson L, et al. Polyclonality of familial murine adenomas: analyses of mouse chimeras with low tumor multiplicity suggest short-range interactions. Proc Natl Acad Sci USA. 2005;102:6960–5.

    Article  CAS  Google Scholar 

  38. Thliveris AT, Schwefel B, Clipson L, et al. Transformation of epithelial cells through recruitment leads to polyclonal intestinal tumors. Proc Natl Acad Sci USA. 2013;110:11523–8.

    Article  Google Scholar 

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Acknowledgements

The authors thank Dr. William F. Goldman for editing the manuscript and Ms. Mutsumi Toyota, Ms. Tomo Hatahira and Ms. Tokiko Ito for technical assistance.

Funding

This study was supported in part by Grant-in-Aid for Scientific Research (C) from the Japan Society for Promotion of Science (JSPS KAKENHI 15K08973, E. Yamamoto), Grant-in-Aid for Scientific Research (B) from the Japan Society for Promotion of Science (JSPS KAKENHI 15H04299, H. Suzuki), Grants-in-Aid for Young Investigators (B) from Japan Society for Promotion of Science (JSPS KAKENHI 15K18431, T. Harada; JSPS KAKENHI 15K19339, H. Aoki), JFE (The Japanese Foundation for Research and Promotion of Endoscopy) Grant (E. Yamamoto), The Takeda Science Foundation (E. Yamamoto) and Suhara Memorial Foundation (E. Yamamoto).

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Author notes

  1. Taku Harada and Eiichiro Yamamoto have contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Biology, Sapporo Medical University School of Medicine, S1, W17, Chuo-ku, Sapporo, 060-8556, Japan

    Taku Harada, Eiichiro Yamamoto, Hironori Aoki, Akira Yorozu, Hiroshi Kitajima, Takeshi Niinuma, Masahiro Kai & Hiromu Suzuki

  2. Center for Gastroenterology, Teine-Keijinkai Hospital, Sapporo, Japan

    Taku Harada & Hironori Aoki

  3. Department of Gastroenterology and Hepatology, Sapporo Medical University School of Medicine, Sapporo, Japan

    Eiichiro Yamamoto, Hiro-o Yamano, Gota Sudo & Hiroshi Nakase

  4. Department of Gastroenterology, Akita Red Cross Hospital, Akita, Japan

    Hiro-o Matsushita, Kenjiro Yoshikawa, Ryo Takagi, Eiji Harada, Yoshihito Tanaka & Yuko Yoshida

  5. Department of Molecular Diagnostic Pathology, Iwate Medical University, Morioka, Japan

    Makoto Eizuka & Tamotsu Sugai

  6. Department of Medical Genome Science, Research Institute for Frontier Medicine, Sapporo Medical University School of Medicine, Sapporo, Japan

    Yasushi Sasaki & Takashi Tokino

Authors
  1. Taku Harada
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  2. Eiichiro Yamamoto
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  3. Hiro-o Yamano
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Corresponding author

Correspondence to Hiromu Suzuki.

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Harada, T., Yamamoto, E., Yamano, Ho. et al. Surface microstructures are associated with mutational intratumoral heterogeneity in colorectal tumors. J Gastroenterol 53, 1241–1252 (2018). https://doi.org/10.1007/s00535-018-1481-z

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  • DOI: https://doi.org/10.1007/s00535-018-1481-z

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