Skip to main content
SpringerLink
Log in

Highly sensitive detection of TERT promoter mutations in recurrent glioblastomas using digital PCR

  • Rapid Short communication
  • Published:
Brain Tumor Pathology Aims and scope Submit manuscript

Abstract

Telomerase reverse transcriptase promoter (TERTp) hotspot mutations are the most frequent mutations in primary glioblastomas (GBM). Previous studies have shown that the combination of TERTp and isocitrate dehydrogenase (IDH) status may serve as a useful diagnostic marker for oligodendroglioma and glioblastoma. In oligodendrogliomas, TERTp and IDH mutations, along with the 1p/19q codeletion, usually coexist and are likely to be founder mutations. However, in contrast to oligodendroglioma, the role of the TERTp status in GBM remains obscure. Here, we used Sanger sequencing, pyrosequencing, and digital PCR (dPCR) to examine the TERTp status in 15 pairs of frozen tissue samples from primary and recurrent IDH wild-type GBM, all of which were operated in a single institute. We showed that the TERTp status was stable between primary and recurrent GBM but this consistency was only detected by dPCR. The results suggest that dPCR is a powerful, highly sensitive tool to detect TERTp mutations, especially in a mixed cell population (e.g., a recurrent GBM tissue) where earlier treatment may have grossly altered the tumor microenvironment.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1

References

  1. Huang FW, Hodis E, Xu MJ, Kryukov GV, Chin L, Garraway LA (2013) Highly recurrent TERT promoter mutations in human melanoma. Science 339:957–959

    Article  CAS  Google Scholar 

  2. Bell RJ, Rube HT, Kreig A, Mancini A, Fouse SD, Nagarajan RP, Choi S, Hong C, He D, Pekmezci M, Wiencke JK, Wrensch MR, Chang SM, Walsh KM, Myong S, Song JS, Costello JF (2015) Cancer. The transcription factor GABP selectively binds and activates the mutant TERT promoter in cancer. Science 348:1036–1039

    Article  CAS  Google Scholar 

  3. Arita H, Narita Y, Fukushima S, Tateishi K, Matsushita Y, Yoshida A, Miyakita Y, Ohno M, Collins VP, Kawahara N, Shibui S, Ichimura K (2013) Upregulating mutations in the TERT promoter commonly occur in adult malignant gliomas and are strongly associated with total 1p19q loss. Acta Neuropathol 126:267–276

    Article  CAS  Google Scholar 

  4. Killela PJ, Reitman ZJ, Jiao Y, Bettegowda C, Agrawal N, Diaz LA, Friedman AH, Friedman H, Gallia GL, Giovanella BC (2013) TERT promoter mutations occur frequently in gliomas and a subset of tumors derived from cells with low rates of self-renewal. Proc Natl Acad Sci 110:6021–6026

    Article  CAS  Google Scholar 

  5. Arita H, Yamasaki K, Matsushita Y, Nakamura T, Shimokawa A, Takami H, Tanaka S, Mukasa A, Shirahata M, Shimizu S, Suzuki K, Saito K, Kobayashi K, Higuchi F, Uzuka T, Otani R, Tamura K, Sumita K, Ohno M, Miyakita Y, Kagawa N, Hashimoto N, Hatae R, Yoshimoto K, Shinojima N, Nakamura H, Kanemura Y, Okita Y, Kinoshita M, Ishibashi K, Shofuda T, Kodama Y, Mori K, Tomogane Y, Fukai J, Fujita K, Terakawa Y, Tsuyuguchi N, Moriuchi S, Nonaka M, Suzuki H, Shibuya M, Maehara T, Saito N, Nagane M, Kawahara N, Ueki K, Yoshimine T, Miyaoka E, Nishikawa R, Komori T, Narita Y, Ichimura K (2016) A combination of TERT promoter mutation and MGMT methylation status predicts clinically relevant subgroups of newly diagnosed glioblastomas. Acta Neuropathol Commun 4:79

    Article  Google Scholar 

  6. Eckel-Passow JE, Lachance DH, Molinaro AM, Walsh KM, Decker PA, Sicotte H, Pekmezci M, Rice T, Kosel ML, Smirnov IV (2015) Glioma groups based on 1p/19q, IDH, and TERT promoter mutations in tumors. N Engl J Med 372:2499–2508

    Article  CAS  Google Scholar 

  7. Suzuki H, Aoki K, Chiba K, Sato Y, Shiozawa Y, Shiraishi Y, Shimamura T, Niida A, Motomura K, Ohka F, Yamamoto T, Tanahashi K, Ranjit M, Wakabayashi T, Yoshizato T, Kataoka K, Yoshida K, Nagata Y, Sato-Otsubo A, Tanaka H, Sanada M, Kondo Y, Nakamura H, Mizoguchi M, Abe T, Muragaki Y, Watanabe R, Ito I, Miyano S, Natsume A, Ogawa S (2015) Mutational landscape and clonal architecture in grade II and III gliomas. Nat Genet 47:458–468

    Article  CAS  Google Scholar 

  8. Killela PJ, Pirozzi CJ, Healy P, Reitman ZJ, Lipp E, Rasheed BA, Yang R, Diplas BH, Wang Z, Greer PK (2014) Mutations in IDH1, IDH2, and in the TERT promoter define clinically distinct subgroups of adult malignant gliomas. Oncotarget 5:1515

    Article  Google Scholar 

  9. Arita H, Yamasaki K, Matsushita Y, Nakamura T, Shimokawa A, Takami H, Tanaka S, Mukasa A, Shirahata M, Shimizu S (2016) A combination of TERT promoter mutation and MGMT methylation status predicts clinically relevant subgroups of newly diagnosed glioblastomas. Acta neuropathologica communications 4:79

    Article  Google Scholar 

  10. Wang J, Cazzato E, Ladewig E, Frattini V, Rosenbloom DI, Zairis S, Abate F, Liu Z, Elliott O, Shin YJ, Lee JK, Lee IH, Park WY, Eoli M, Blumberg AJ, Lasorella A, Nam DH, Finocchiaro G, Iavarone A, Rabadan R (2016) Clonal evolution of glioblastoma under therapy. Nat Genet 48:768–776

    Article  CAS  Google Scholar 

  11. Lee JK, Wang J, Sa JK, Ladewig E, Lee HO, Lee IH, Kang HJ, Rosenbloom DS, Camara PG, Liu Z, van Nieuwenhuizen P, Jung SW, Choi SW, Kim J, Chen A, Kim KT, Shin S, Seo YJ, Oh JM, Shin YJ, Park CK, Kong DS, Seol HJ, Blumberg A, Lee JI, Iavarone A, Park WY, Rabadan R, Nam DH (2017) Spatiotemporal genomic architecture informs precision oncology in glioblastoma. Nat Genet 49:594–599

    Article  CAS  Google Scholar 

  12. Kim J, Lee IH, Cho HJ, Park CK, Jung YS, Kim Y, Nam SH, Kim BS, Johnson MD, Kong DS, Seol HJ, Lee JI, Joo KM, Yoon Y, Park WY, Lee J, Park PJ, Nam DH (2015) Spatiotemporal evolution of the primary glioblastoma genome. Cancer Cell 28:318–328

    Article  CAS  Google Scholar 

  13. Johnson BE, Mazor T, Hong C, Barnes M, Aihara K, McLean CY, Fouse SD, Yamamoto S, Ueda H, Tatsuno K (2014) Mutational analysis reveals the origin and therapy-driven evolution of recurrent glioma. Science 343:189–193

    Article  CAS  Google Scholar 

  14. Lee JH, Lee JE, Kahng JY, Kim SH, Park JS, Yoon SJ, Um J-Y, Kim WK, Lee J-K, Park J (2018) Human glioblastoma arises from subventricular zone cells with low-level driver mutations. Nature 560:243

    Article  CAS  Google Scholar 

  15. Körber V, Yang J, Barah P, Wu Y, Stichel D, Gu Z, Fletcher MNC, Jones D, Hentschel B, Lamszus K (2019) Evolutionary trajectories of IDHWT glioblastomas reveal a common path of early tumorigenesis instigated years ahead of initial diagnosis. Cancer Cell 35:692–704.e612

    Article  Google Scholar 

  16. Zhang Z, Chan AK-Y, Ding X, Li Y, Zhang R, Chen L, Liu Y, Wang Y, Xiong J, Ng H-K (2017) Glioma groups classified by IDH and TERT promoter mutations remain stable among primary and recurrent gliomas. Neuro Oncology 19:1008–1010

    Article  Google Scholar 

  17. Abou-El-Ardat K, Seifert M, Becker K, Eisenreich S, Lehmann M, Hackmann K, Rump A, Meijer G, Carvalho B, Temme A (2017) Comprehensive molecular characterization of multifocal glioblastoma proves its monoclonal origin and reveals novel insights into clonal evolution and heterogeneity of glioblastomas. Neuro-oncology 19:546–557

    Article  CAS  Google Scholar 

  18. Diplas BH, Liu H, Yang R, Hansen LJ, Zachem AL, Zhao F, Bigner DD, McLendon RE, Jiao Y, He Y (2018) Sensitive and rapid detection of TERT promoter and IDH mutations in diffuse gliomas. Neuro Oncol 21:440–450

    Article  Google Scholar 

  19. Brat DJ, Aldape K, Colman H, Holland EC, Louis DN, Jenkins RB, Kleinschmidt-DeMasters B, Perry A, Reifenberger G, Stupp R (2018) cIMPACT-NOW update 3: recommended diagnostic criteria for “diffuse astrocytic glioma, IDH-wildtype, with molecular features of glioblastoma, WHO grade IV”. Acta Neuropathol 136:805–810

    Article  CAS  Google Scholar 

Download references

Acknowledgments

K.I. was supported by the Japan Cancer Research Project and Practical Research for Innovative Cancer Control programs from the Japan Agency for Medical Research and Development (AMED).

Author information

Authors and Affiliations

  1. Division of Brain Tumor Translational Research, National Cancer Center Research Institute, Tokyo, Japan

    Shunichiro Miki, Kaishi Satomi, Yuko Matsushita, Mai Kitahara & Koichi Ichimura

  2. Department of Neurosurgery and Neuro-Oncology, National Cancer Center Hospital, Tokyo, Japan

    Shunichiro Miki, Makoto Ohno, Yuko Matsushita, Yasuji Miyakita, Masamichi Takahashi & Yoshitaka Narita

  3. Department of Pathology and Clinical Laboratories, National Cancer Center Hospital, Tokyo, Japan

    Kaishi Satomi & Akihiko Yoshida

  4. Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki, Japan

    Shunichiro Miki, Masahide Matsuda, Eiichi Ishikawa & Akira Matsumura

Authors
  1. Shunichiro Miki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kaishi Satomi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Makoto Ohno
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yuko Matsushita
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mai Kitahara
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yasuji Miyakita
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Masamichi Takahashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Masahide Matsuda
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Eiichi Ishikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Akira Matsumura
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Akihiko Yoshida
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Yoshitaka Narita
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Koichi Ichimura
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Koichi Ichimura.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest related to this work.

Ethical approval and informed consent

All procedures involving patients were performed in accordance with the ethical standards of the National Cancer Center and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed consent was obtained to use tumor material according to the procedures outlined by the National Cancer Center.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Miki, S., Satomi, K., Ohno, M. et al. Highly sensitive detection of TERT promoter mutations in recurrent glioblastomas using digital PCR. Brain Tumor Pathol 37, 154–158 (2020). https://doi.org/10.1007/s10014-020-00375-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10014-020-00375-x

Keywords

Search

Navigation