Detection of oncogenic mutations in paired circulating tumor DNA and circulating tumor cells in patients with hepatocellular carcinoma

Zhouhong Ge; Jean C.A. Helmijr; Maurice P.H.M. Jansen; Patrick P.C. Boor; Lisanne Noordam; Maikel Peppelenbosch; Jaap Kwekkeboom; Jaco Kraan; Dave Sprengers

doi:10.1016/j.tranon.2021.101073

Detection of oncogenic mutations in paired circulating tumor DNA and circulating tumor cells in patients with hepatocellular carcinoma

Zhouhong Ge
, Jean C.A. Helmijr
, Maurice P.H.M. Jansen
, Patrick P.C. Boor
, Lisanne Noordam
, Maikel Peppelenbosch
, Jaap Kwekkeboom
, Jaco Kraan
, Dave Sprengers^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

20 Citations (Scopus)

Abstract

Background and aims: Circulating tumor cells (CTCs) or circulating tumor DNA (ctDNA) may be used for diagnostic or prognostic purposes in patients with hepatocellular carcinoma (HCC). We aim to determine whether CTCs or ctDNA are suitable to determine oncogenic mutations in HCC patients. Methods: Twenty-six mostly advanced HCC patients were enrolled. 30 mL peripheral blood from each patient was obtained. CellSearch system was used for CTC detection. A sequencing panel covering 14 cancer-relevant genes was used to identify oncogenic mutations. TERT promoter C228T and C250T mutations were determined by droplet digital PCR. Results: CTCs were detected in 27% (7/26) of subjects but at low numbers (median: 2 cells, range: 1–15 cells) and ctDNA in 77% (20/26) of patients. Mutations in ctDNA were identified in several genes: TERT promoter C228T (77%, 20/26), TP53 (23%, 6/26), CTNNB1 (12%, 3/26), PIK3CA (12%, 3/26) and NRAS (4%, 1/26). The TERT C228T mutation was present in all patients with one or more ctDNA mutations, or detectable CTCs. The TERT C228T and TP53 mutations detected in ctDNA were present at higher levels in matched primary HCC tumor tissue. The maximal variant allele frequency (VAF) of ctDNA was linearly correlated with largest tumor size and AFP level (Log10). CtDNA (or TERT C228T) positivity was associated with macrovascular invasion, and positivity of ctDNA (or TERT C228T) or CTCs (≥ 2) correlated with poor patient survival. Conclusions: Oncogenic mutations could be detected in ctDNA from advanced HCC patients. CtDNA analysis may serve as a promising liquid biopsy to identify druggable mutations.

Original language	English
Article number	101073
Journal	Translational Oncology
Volume	14
Issue number	7
DOIs	https://doi.org/10.1016/j.tranon.2021.101073
Publication status	Published - Jul 2021

Bibliographical note

Funding Information:
This study was supported by the China Scholarship Council which provided a Ph.D fellowship grant to Zhouhong Ge (number 201606230253) and a Gastrostart fund provided to Dave Sprengers by the Dutch Society for Gastroenterology (NVGE; BO/JG/PROJECT 2016-10).

Funding Information:
We would like to thank Corine Beaufort and Mai Van for technical support (Department of Medical Oncology). This study was supported by the China Scholarship Council which provided a Ph.D fellowship grant to Zhouhong Ge (number 201606230253) and a Gastrostart fund provided to Dave Sprengers by the Dutch Society for Gastroenterology (NVGE; BO/JG/PROJECT 2016-10). The authors declare no potential conflict of interest. No, ZG, JKw, JKr and DS conceived the idea and designed the study. ZG processed patient samples, analyzed data and wrote the draft of the manuscript. JKw, JKr and DS supervised the study. JH and PB assisted with experiments. JH and MJ contributed to ddPCR and NGS data analysis. DS contributed to material support. JH, MJ, LN, JKw, JKr and DS contributed to revision of the manuscript. ZG and DS obtained funding. The study was approved by the local ethics committee (METC), Erasmus MC, Rotterdam (METC number: 17-238), and written informed consent was obtained from each patient. The data will be made available upon reasonable request.

Publisher Copyright:
© 2021

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10.1016/j.tranon.2021.101073

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Ge, Z., Helmijr, J. C. A., Jansen, M. P. H. M., Boor, P. P. C., Noordam, L., Peppelenbosch, M., Kwekkeboom, J., Kraan, J., & Sprengers, D. (2021). Detection of oncogenic mutations in paired circulating tumor DNA and circulating tumor cells in patients with hepatocellular carcinoma. Translational Oncology, 14(7), Article 101073. https://doi.org/10.1016/j.tranon.2021.101073

@article{f09eae7bd4e14d1f9545fea5366a7e37,

title = "Detection of oncogenic mutations in paired circulating tumor DNA and circulating tumor cells in patients with hepatocellular carcinoma",

abstract = "Background and aims: Circulating tumor cells (CTCs) or circulating tumor DNA (ctDNA) may be used for diagnostic or prognostic purposes in patients with hepatocellular carcinoma (HCC). We aim to determine whether CTCs or ctDNA are suitable to determine oncogenic mutations in HCC patients. Methods: Twenty-six mostly advanced HCC patients were enrolled. 30 mL peripheral blood from each patient was obtained. CellSearch system was used for CTC detection. A sequencing panel covering 14 cancer-relevant genes was used to identify oncogenic mutations. TERT promoter C228T and C250T mutations were determined by droplet digital PCR. Results: CTCs were detected in 27\% (7/26) of subjects but at low numbers (median: 2 cells, range: 1–15 cells) and ctDNA in 77\% (20/26) of patients. Mutations in ctDNA were identified in several genes: TERT promoter C228T (77\%, 20/26), TP53 (23\%, 6/26), CTNNB1 (12\%, 3/26), PIK3CA (12\%, 3/26) and NRAS (4\%, 1/26). The TERT C228T mutation was present in all patients with one or more ctDNA mutations, or detectable CTCs. The TERT C228T and TP53 mutations detected in ctDNA were present at higher levels in matched primary HCC tumor tissue. The maximal variant allele frequency (VAF) of ctDNA was linearly correlated with largest tumor size and AFP level (Log10). CtDNA (or TERT C228T) positivity was associated with macrovascular invasion, and positivity of ctDNA (or TERT C228T) or CTCs (≥ 2) correlated with poor patient survival. Conclusions: Oncogenic mutations could be detected in ctDNA from advanced HCC patients. CtDNA analysis may serve as a promising liquid biopsy to identify druggable mutations.",

author = "Zhouhong Ge and Helmijr, \{Jean C.A.\} and Jansen, \{Maurice P.H.M.\} and Boor, \{Patrick P.C.\} and Lisanne Noordam and Maikel Peppelenbosch and Jaap Kwekkeboom and Jaco Kraan and Dave Sprengers",

note = "Funding Information: This study was supported by the China Scholarship Council which provided a Ph.D fellowship grant to Zhouhong Ge (number 201606230253) and a Gastrostart fund provided to Dave Sprengers by the Dutch Society for Gastroenterology (NVGE; BO/JG/PROJECT 2016-10). Funding Information: We would like to thank Corine Beaufort and Mai Van for technical support (Department of Medical Oncology). This study was supported by the China Scholarship Council which provided a Ph.D fellowship grant to Zhouhong Ge (number 201606230253) and a Gastrostart fund provided to Dave Sprengers by the Dutch Society for Gastroenterology (NVGE; BO/JG/PROJECT 2016-10). The authors declare no potential conflict of interest. No, ZG, JKw, JKr and DS conceived the idea and designed the study. ZG processed patient samples, analyzed data and wrote the draft of the manuscript. JKw, JKr and DS supervised the study. JH and PB assisted with experiments. JH and MJ contributed to ddPCR and NGS data analysis. DS contributed to material support. JH, MJ, LN, JKw, JKr and DS contributed to revision of the manuscript. ZG and DS obtained funding. The study was approved by the local ethics committee (METC), Erasmus MC, Rotterdam (METC number: 17-238), and written informed consent was obtained from each patient. The data will be made available upon reasonable request. Publisher Copyright: {\textcopyright} 2021",

year = "2021",

month = jul,

doi = "10.1016/j.tranon.2021.101073",

language = "English",

volume = "14",

journal = "Translational Oncology",

issn = "1936-5233",

publisher = "Neoplasia Press, Inc.",

number = "7",

}

TY - JOUR

T1 - Detection of oncogenic mutations in paired circulating tumor DNA and circulating tumor cells in patients with hepatocellular carcinoma

AU - Ge, Zhouhong

AU - Helmijr, Jean C.A.

AU - Jansen, Maurice P.H.M.

AU - Boor, Patrick P.C.

AU - Noordam, Lisanne

AU - Peppelenbosch, Maikel

AU - Kwekkeboom, Jaap

AU - Kraan, Jaco

AU - Sprengers, Dave

N1 - Funding Information: This study was supported by the China Scholarship Council which provided a Ph.D fellowship grant to Zhouhong Ge (number 201606230253) and a Gastrostart fund provided to Dave Sprengers by the Dutch Society for Gastroenterology (NVGE; BO/JG/PROJECT 2016-10). Funding Information: We would like to thank Corine Beaufort and Mai Van for technical support (Department of Medical Oncology). This study was supported by the China Scholarship Council which provided a Ph.D fellowship grant to Zhouhong Ge (number 201606230253) and a Gastrostart fund provided to Dave Sprengers by the Dutch Society for Gastroenterology (NVGE; BO/JG/PROJECT 2016-10). The authors declare no potential conflict of interest. No, ZG, JKw, JKr and DS conceived the idea and designed the study. ZG processed patient samples, analyzed data and wrote the draft of the manuscript. JKw, JKr and DS supervised the study. JH and PB assisted with experiments. JH and MJ contributed to ddPCR and NGS data analysis. DS contributed to material support. JH, MJ, LN, JKw, JKr and DS contributed to revision of the manuscript. ZG and DS obtained funding. The study was approved by the local ethics committee (METC), Erasmus MC, Rotterdam (METC number: 17-238), and written informed consent was obtained from each patient. The data will be made available upon reasonable request. Publisher Copyright: © 2021

PY - 2021/7

Y1 - 2021/7

N2 - Background and aims: Circulating tumor cells (CTCs) or circulating tumor DNA (ctDNA) may be used for diagnostic or prognostic purposes in patients with hepatocellular carcinoma (HCC). We aim to determine whether CTCs or ctDNA are suitable to determine oncogenic mutations in HCC patients. Methods: Twenty-six mostly advanced HCC patients were enrolled. 30 mL peripheral blood from each patient was obtained. CellSearch system was used for CTC detection. A sequencing panel covering 14 cancer-relevant genes was used to identify oncogenic mutations. TERT promoter C228T and C250T mutations were determined by droplet digital PCR. Results: CTCs were detected in 27% (7/26) of subjects but at low numbers (median: 2 cells, range: 1–15 cells) and ctDNA in 77% (20/26) of patients. Mutations in ctDNA were identified in several genes: TERT promoter C228T (77%, 20/26), TP53 (23%, 6/26), CTNNB1 (12%, 3/26), PIK3CA (12%, 3/26) and NRAS (4%, 1/26). The TERT C228T mutation was present in all patients with one or more ctDNA mutations, or detectable CTCs. The TERT C228T and TP53 mutations detected in ctDNA were present at higher levels in matched primary HCC tumor tissue. The maximal variant allele frequency (VAF) of ctDNA was linearly correlated with largest tumor size and AFP level (Log10). CtDNA (or TERT C228T) positivity was associated with macrovascular invasion, and positivity of ctDNA (or TERT C228T) or CTCs (≥ 2) correlated with poor patient survival. Conclusions: Oncogenic mutations could be detected in ctDNA from advanced HCC patients. CtDNA analysis may serve as a promising liquid biopsy to identify druggable mutations.

AB - Background and aims: Circulating tumor cells (CTCs) or circulating tumor DNA (ctDNA) may be used for diagnostic or prognostic purposes in patients with hepatocellular carcinoma (HCC). We aim to determine whether CTCs or ctDNA are suitable to determine oncogenic mutations in HCC patients. Methods: Twenty-six mostly advanced HCC patients were enrolled. 30 mL peripheral blood from each patient was obtained. CellSearch system was used for CTC detection. A sequencing panel covering 14 cancer-relevant genes was used to identify oncogenic mutations. TERT promoter C228T and C250T mutations were determined by droplet digital PCR. Results: CTCs were detected in 27% (7/26) of subjects but at low numbers (median: 2 cells, range: 1–15 cells) and ctDNA in 77% (20/26) of patients. Mutations in ctDNA were identified in several genes: TERT promoter C228T (77%, 20/26), TP53 (23%, 6/26), CTNNB1 (12%, 3/26), PIK3CA (12%, 3/26) and NRAS (4%, 1/26). The TERT C228T mutation was present in all patients with one or more ctDNA mutations, or detectable CTCs. The TERT C228T and TP53 mutations detected in ctDNA were present at higher levels in matched primary HCC tumor tissue. The maximal variant allele frequency (VAF) of ctDNA was linearly correlated with largest tumor size and AFP level (Log10). CtDNA (or TERT C228T) positivity was associated with macrovascular invasion, and positivity of ctDNA (or TERT C228T) or CTCs (≥ 2) correlated with poor patient survival. Conclusions: Oncogenic mutations could be detected in ctDNA from advanced HCC patients. CtDNA analysis may serve as a promising liquid biopsy to identify druggable mutations.

UR - https://www.scopus.com/pages/publications/85108459490

U2 - 10.1016/j.tranon.2021.101073

DO - 10.1016/j.tranon.2021.101073

M3 - Article

AN - SCOPUS:85108459490

SN - 1936-5233

VL - 14

JO - Translational Oncology

JF - Translational Oncology

IS - 7

M1 - 101073

ER -

Detection of oncogenic mutations in paired circulating tumor DNA and circulating tumor cells in patients with hepatocellular carcinoma

Abstract

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