Skip to main content
SpringerLink
Log in

Liquid Biopsy im nicht-kleinzelligen Lungenkarzinom

Die blutbasierte Analyse der ctDNA-Methylierung

Liquid biopsy in human non-small-cell lung cancer

Blood-based analysis of ctDNA methylation

  • Referate Preisträger: Promotionspreis
  • Published:
Der Pathologe Aims and scope Submit manuscript

Zusammenfassung

Hintergrund

Einsatz der Liquid Biopsy als minimalinvasive Methode für die Nachsorgediagnostik des nicht-kleinzelligen Lungenkarzinoms (NSCLC).

Fragestellung

Systematische Suche nach neuen blutbasierten DNA-Methylierungsmarkern zur Differenzierung von NSCLC-Patienten und Patienten ohne maligne Erkrankung.

Material und Methode

Quantitative Analyse der DNA-Methylierung in der Promotorregion potenzieller Biomarkergene aus cfDNA (Plasma) mittels Pyrosequenzierung.

Ergebnisse

Die Untersuchung der cfDNA-Hypermethylierung aus Plasma ergab für den Biomarker CFTR eine signifikant höhere Methylierung bei NSCLC-Patienten verglichen mit allen Kontrollen und der NSCLC-Patientengruppe nach kurativer Behandlung eines primären Lungenkarzinoms. Eine ROC-Analyse der am besten diskriminierenden CpGs des CFTR-Promotors (CpG1-2-4) erreichte eine Sensitivität von 52 % bei einer Spezifität von 90 % im Vergleich von NSCLC-Patienten und Post-NSCLC-Patienten (AUC = 0,69, p-Wert < 0,05).

Schlussfolgerung

Die Promotor Hypermethylierung des potenziellen Biomarkers CFTR zeigt eine gute Tendenz zur Differenzierung von NSCLC-Patienten und Post-NSCLC-Patienten und sollte weiter evaluiert werden.

Abstract

Background

Use of liquid biopsy for minimal invasive follow-up diagnostics of non-small-cell lung carcinomas (NSCLCs).

Objectives

Systematic search for new putative blood-based hypermethylation biomarkers to discriminate NSCLC patients from patients without a malign disease.

Methods

Quantitative analysis of gene promoter DNA methylation of potential biomarkers from cfDNA (plasma) with pyrosequencing.

Results

cfDNA hypermethylation in plasma confirmed significant higher methylation frequencies of the candidate gene CFTR of the NSCLC patients compared to the combined control groups and to NSCLC patients after curative therapy of primary NSCLC (post-NSCLC). ROC-analysis of the best discriminatory CpGs of the CFTR promotor (CpG1-2-4) revealed a sensitivity of 52% in NSCLC patients and a specificity of 90% in the post-NSCLC group (AUC: 0.69; p < 0.05).

Conclusions

Promotor hypermethylation of the potential biomarker CFTR shows a discriminatory potential for differentiation of NSCLC patients to patients without a malign disease and should further be investigated.

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.

Institutional subscriptions

Abb. 1
Abb. 2
Abb. 3
Abb. 4
Abb. 5
Abb. 6

Literatur

  1. Diaz LA Jr et al (2012) The molecular evolution of acquired resistance to targeted EGFR blockade in colorectal cancers. Nature 486(7404):537–540

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Mandel P (1948) Les acides nucleiques du plasma sanguin chez l’homme. Cr Acad Sci Paris 142:241–243

    CAS  Google Scholar 

  3. Diehl F et al (2005) Detection and quantification of mutations in the plasma of patients with colorectal tumors. Proc Natl Acad Sci USA 102(45):16368–16373

    Article  CAS  PubMed  Google Scholar 

  4. Stroun M et al (1989) Neoplastic characteristics of the DNA found in the plasma of cancer patients. Oncology 46(5):318–322

    Article  CAS  PubMed  Google Scholar 

  5. Diehl F et al (2008) Circulating mutant DNA to assess tumor dynamics. Nat Med 14(9):985–990

    Article  CAS  PubMed  Google Scholar 

  6. Stroun M et al (2001) About the possible origin and mechanism of circulating DNA: Apoptosis and active DNA release. Clin Chim Acta 313(1):139–142

    Article  CAS  PubMed  Google Scholar 

  7. Norton S et al (2013) A stabilizing reagent prevents cell-free DNA contamination by cellular DNA in plasma during blood sample storage and shipping as determined by digital PCR. Clin Biochem 46(15):1561–1565

    Article  CAS  PubMed  Google Scholar 

  8. Baylin SB, Jones PA (2011) A decade of exploring the cancer epigenome—biological and translational implications. Nat Rev Cancer 11(10):726–734

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Baylin SB, Herman JG (2000) DNA hypermethylation in tumorigenesis: epigenetics joins genetics. Trends Genet 16(4):168–174

    Article  CAS  PubMed  Google Scholar 

  10. Esteller M, Herman JG (2002) Cancer as an epigenetic disease: DNA methylation and chromatin alterations in human tumours. J Pathol 196(1):1–7

    Article  CAS  PubMed  Google Scholar 

  11. Bach PB et al (2012) Benefits and harms of CT screening for lung cancer: a systematic review. JAMA 307(22):2418–2429

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Wender R et al (2013) American Cancer Society lung cancer screening guidelines. CA Cancer J Clin 63(2):106–117

    Article  Google Scholar 

  13. Goeckenjan G et al (2011) Prevention, diagnosis, therapy, and follow-up of lung cancer. Pneumologie 65(01):39–59

    Article  CAS  PubMed  Google Scholar 

  14. Aberle DR et al (2013) Results of the two incidence screenings in the National Lung Screening Trial. N Engl J Med 369(10):920–931

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Patz EF et al (2014) Overdiagnosis in low-dose computed tomography screening for lung cancer. JAMA Intern Med 174(2):269–274

    Article  PubMed  PubMed Central  Google Scholar 

  16. Kozower BD et al (2010) STS database risk models: predictors of mortality and major morbidity for lung cancer resection. Ann Thorac Surg 90(3):875–883

    Article  PubMed  Google Scholar 

  17. Balak MN et al (2006) Novel D761Y and common secondary T790M mutations in epidermal growth factor receptor-mutant lung adenocarcinomas with acquired resistance to kinase inhibitors. Clin Cancer Res 12(21):6494–6501

    Article  CAS  PubMed  Google Scholar 

  18. Yu H et al (2013) Analysis of Mechanisms of Acquired Resistance to EGFR TKI therapy in 155 patients with EGFR-mutant Lung Cancers. Clin Cancer Res. https://doi.org/10.1158/1078-0432.CCR-12-2246

    Article  PubMed  PubMed Central  Google Scholar 

  19. Kimura H et al (2006) Detection of epidermal growth factor receptor mutations in serum as a predictor of the response to gefitinib in patients with non-small-cell lung cancer. Clin Cancer Res 12(13):3915–3921

    Article  CAS  PubMed  Google Scholar 

  20. AWMF, D. Krebshilfe, D. Krebsgesellschaft (2018) S3-Leitlinie: Prävention, Diagnostik, Therapie und Nachsorge des Lungenkarzinoms. www.awmf.org

    Google Scholar 

  21. Oxnard GR et al (2014) Noninvasive detection of response and resistance in EGFR-mutant lung cancer using quantitative next-generation genotyping of cell-free plasma DNA. Clin Cancer Res 20(6):1698–1705

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Frangioni JV (2008) New technologies for human cancer imaging. J Clin Oncol 26(24):4012–4021

    Article  PubMed  PubMed Central  Google Scholar 

  23. Berlin K, Ballhause M, Cardon K (2004) Bisulfite conversion of DNA. Google Patents, https://patents.google.com/patent/US20060134643A1/en

  24. Delaney C, Garg SK, Yung R (2015) Analysis of DNA Methylation by Pyrosequencing. In: Shaw A (Hrsg) Immunosenescence. Methods in Molecular Biology, Bd. 1343. Humana Press, New York, S 249–264

    Google Scholar 

  25. Nelder JA, Baker RJ (2004) Generalized linear models. Encyclopedia of statistical sciences, Bd. 4

    Google Scholar 

  26. Hochberg Y (1988) A sharper Bonferroni procedure for multiple tests of significance. Biometrika 75(4):800–802

    Article  Google Scholar 

  27. Son JW et al (2011) Promoter hypermethylation of the CFTR gene and clinical/pathological features associated with non-small cell lung cancer. Respirology 16(8):1203–1209

    Article  PubMed  Google Scholar 

  28. Kaedbey R et al (2015) Noninvasive diagnosis of actionable mutations by deep sequencing of circulating cell free DNA (cfDNA) in multiple myeloma (MM). Clin Lymphoma Myeloma Leuk 15:e45–e46

    Article  Google Scholar 

  29. Paweletz CP et al (2016) Bias-corrected targeted next-generation sequencing for rapid, multiplexed detection of actionable alterations in cell-free DNA from advanced lung cancer patients. Clin Cancer Res 22(4):915–922

    Article  CAS  PubMed  Google Scholar 

  30. Day E, Dear PH, McCaughan F (2013) Digital PCR strategies in the development and analysis of molecular biomarkers for personalized medicine. Methods 59(1):101–107

    Article  CAS  PubMed  Google Scholar 

  31. Wiencke JK et al (2014) A comparison of DNA methylation specific droplet digital PCR (ddPCR) and real time qPCR with flow cytometry in characterizing human T cells in peripheral blood. Epigenetics 9(10):1360–1365

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Danksagung

Die Autorin dankt Frau Dr. rer. nat. Vera Kloten und Herrn Professor Dr. rer. nat. Edgar Dahl für die Betreuung der Promotionsarbeit.

Author information

Authors and Affiliations

  1. Arbeitsgruppe Molekulare Onkologie, Institut für Pathologie, Uniklinik RWTH Aachen, Pauwelsstraße 30, 52074, Aachen, Deutschland

    H. Schulz, M. Tator, R. Knüchel-Clarke,  V. Kloten &  E. Dahl

  2. Klinik für Thorax‑, Herz- und Gefäßchirurgie, Uniklinik RWTH Aachen, Aachen, Deutschland

    J. Spillner

  3. Klinik für Pneumologie und Internistische Intensivmedizin, Uniklinik RWTH Aachen, Aachen, Deutschland

    M. Dreher

  4. RWTH zentralisierte Biomaterialbank (RWTH cBMB) am Institut für Pathologie, Uniklinik RWTH Aachen, Aachen, Deutschland

    E. Dahl

Authors
  1. H. Schulz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Tator
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. Spillner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. Dreher
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. R. Knüchel-Clarke
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. V. Kloten
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. E. Dahl
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to H. Schulz.

Ethics declarations

Interessenkonflikt

H. Schulz, M. Tator, J. Spillner, M. Dreher, R. Knüchel-Clarke, V. Kloten und E. Dahl geben an, dass kein Interessenkonflikt besteht.

Alle beschriebenen Untersuchungen am Menschen wurden mit Zustimmung der zuständigen Ethik-Kommission, im Einklang mit nationalem Recht sowie gemäß der Deklaration von Helsinki von 1975 (in der aktuellen, überarbeiteten Fassung) durchgeführt. Von allen beteiligten Patienten liegt eine Einverständniserklärung vor.

The supplement containing this article is not sponsored by industry.

Additional information

Bei diesem Beitrag handelt es sich um einen Teil der Dissertation von Frau Hanna Schulz. Die Arbeit dient der Erfüllung von Voraussetzungen zur Erlangung des medizinischen Doktorgrades an der Rheinisch-Westfälischen Technischen Hochschule (RWTH) Aachen. Die Autoren V. Kloten und E. Dahl haben zu gleichen Teilen zu der Arbeit beigetragen.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Schulz, H., Tator, M., Spillner, J. et al. Liquid Biopsy im nicht-kleinzelligen Lungenkarzinom. Pathologe 39 (Suppl 2), 193–198 (2018). https://doi.org/10.1007/s00292-018-0536-5

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00292-018-0536-5

Schlüsselwörter

Keywords

Search

Navigation