Skip to main content
SpringerLink
Log in

Liquid Biopsy im kolorektalen Karzinom

Ein Überblick zur ctDNA-Analyse in der Tumordiagnostik

Liquid biopsy in colorectal cancer

An overview of ctDNA analysis in tumour diagnostics

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

Zusammenfassung

Die routinemäßige molekulare Charakterisierung von Kolonkarzinomen erfolgt an Biopsien oder Resektaten. Dieser Goldstandard zur Klassifikation des Tumors kann jedoch die Tumorheterogenität nicht vollständig repräsentieren, denn eine Gewebeprobe stellt nur eine regionale Momentaufnahme dar. Zur Bestimmung des Mutationsprofils potenziell aller Tumorherde sowie zur Überprüfung des Mutationsstatus in regelmäßigen Abständen kann die minimal-invasive Liquid-Biopsy-Diagnostik verwendet werden. Dabei wird die im Blut zirkulierende, zellfreie DNA von Tumorzellen („circulating cell-free tumour DNA“, ctDNA) isoliert, die von apoptotischen oder nekrotischen Zellen freigesetzt oder aktiv von zirkulierenden Tumorzellen („circulating tumour cells“, CTCs) ins Blut abgegeben wird. Die Herausforderung bei der Analyse besteht in der geringen Menge an ctDNA im BIut und deren starker Fragmentierung sowie dem Vorliegen eines hohen Wildtyphintergrundes aufgrund von zellfreier DNA („cell-free DNA“, cfDNA), die von gesunden Zellen abgegeben wird. Dieser Übersichtsartikel beschreibt die Anwendungsfelder, das Potenzial und die Herausforderungen der ctDNA-Analyse im Allgemeinen und insbesondere in Bezug auf einen Einsatz zur Tumordiagnostik von kolorektalen Karzinomen (CRCs). Dabei wird besonders die Anwendung der Liquid Biopsy bei CRC-Patienten unter Anti-EGFR-Therapie diskutiert, weil das Monitoring zur Detektion von Resistenzmutationen (z. B. KRAS-Mutationen) von klinischer Relevanz ist. Außerdem wird die Konkordanz der gewebe- und blutbasierten Tumor-DNA-Analyse betrachtet und diskutiert, ob und inwieweit die Methodik der Liquid Biopsy den molekularpathologischen Standard der gewebebasierten DNA-Analyse zukünftig ergänzen kann.

Abstract

In current routine diagnostics, the gold standard to determine the genomic profile of colorectal cancers (CRCs) is using biopsy or surgically resected tissues. However, such a tissue sample cannot represent the entire tumour heterogeneity, because it only shows a local and temporal snapshot. As a complement to tumour tissue genotyping, liquid biopsies enable minimally invasive detection of all potential tumour-specific mutations and their dynamic changes for molecular profiling. Furthermore, they can be repeated in certain intervals for monitoring response to treatment, occurrence of drug resistance and detection of relapse. This review focusses on analyzing circulating cell-free tumour DNA (ctDNA), which is mostly released from apoptotic or necrotic tumour cells into the bloodstream or by active secretion of circulating tumour cells (CTCs). Nevertheless, there are some challenges in analyzing ctDNA. First, ctDNA represents only a small fraction of total circulating DNA, because there is an enormous wild-type background of cell-free DNA (cfDNA) released by healthy cells. Second, ctDNA is highly fragmented and the amount of ctDNA in the blood is very low. In this review, we discuss the potential, fields of application as well as challenges and limitations of liquid biopsy approaches. In more detail, we discuss the possibility of using liquid biopsies as a future application for molecular characterization of CRCs, particularly for monitoring CRC patients during anti-EGFR therapy to detect resistance mutations (e.g. KRAS mutations) or further therapy-relevant mutations. In addition, we investigate whether blood-based molecular profiling is a reliable addition to routine diagnostic approaches of tissue-based molecular characterization.

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

Literatur

  1. Alix-Panabières C, Pantel K (2017) Clinical prospects of liquid biopsies. Nat Biomed Eng. https://doi.org/10.1038/s41551-017-0065

    Article  Google Scholar 

  2. Bachet JB, Bouche O, Taieb J et al (2018) RAS mutation analysis in circulating tumor DNA from patients with metastatic colorectal cancer: the AGEO RASANC prospective multicenter study. Ann Oncol 29:1211–1219

    CAS  PubMed  Google Scholar 

  3. Baran B, Mert Ozupek N, Yerli Tetik N et al (2018) Difference between left-sided and right-sided colorectal cancer: a focused review of literature. Gastroenterology Res 11:264–273

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Bardelli A, Corso S, Bertotti A et al (2013) Amplification of the MET receptor drives resistance to anti-EGFR therapies in colorectal cancer. Cancer Discov 3:658–673

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Bardelli A, Pantel K (2017) Liquid biopsies, what we do not know (yet). Cancer Cell 31:172–179

    CAS  PubMed  Google Scholar 

  6. Bettegowda C, Sausen M, Leary RJ et al (2014) Detection of circulating tumor DNA in early- and late-stage human malignancies. Sci Transl Med 6:224ra224

    Google Scholar 

  7. Bronkhorst AJ, Ungerer V, Holdenrieder S (2019) The emerging role of cell-free DNA as a molecular marker for cancer management. Biomol Detect Quantif 17:100087

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Church TR, Wandell M, Lofton-Day C et al (2014) Prospective evaluation of methylated SEPT9 in plasma for detection of asymptomatic colorectal cancer. Gut 63:317–325

    CAS  PubMed  Google Scholar 

  9. Cohen JD, Li L, Wang Y et al (2018) Detection and localization of surgically resectable cancers with a multi-analyte blood test. Science 359:926–930

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Corcoran RB, Chabner BA (2018) Application of cell-free DNA analysis to cancer treatment. N Engl J Med 379:1754–1765

    CAS  PubMed  Google Scholar 

  11. Crowley E, Di Nicolantonio F, Loupakis F et al (2013) Liquid biopsy: monitoring cancer-genetics in the blood. Nat Rev Clin Oncol 10:472–484

    CAS  PubMed  Google Scholar 

  12. Dagogo-Jack I, Shaw AT (2018) Tumour heterogeneity and resistance to cancer therapies. Nat Rev Clin Oncol 15:81–94

    CAS  PubMed  Google Scholar 

  13. Diaz LA Jr., Williams RT, Wu J et al (2012) The molecular evolution of acquired resistance to targeted EGFR blockade in colorectal cancers. Nature 486:537–540

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Diaz LA, Marabelle A, Delord J‑P et al (2017) Pembrolizumab therapy for microsatellite instability high (MSI-H) colorectal cancer (CRC) and non-CRC. J Clin Oncol 35:3071–3071

    Google Scholar 

  15. Diehl F, Li M, Dressman D et al (2005) Detection and quantification of mutations in the plasma of patients with colorectal tumors. Proc Natl Acad Sci U S A 102:16368–16373

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Diehl F, Schmidt K, Choti MA et al (2008) Circulating mutant DNA to assess tumor dynamics. Nat Med 14:985–990

    CAS  PubMed  Google Scholar 

  17. Dienstmann R, Vermeulen L, Guinney J et al (2017) Consensus molecular subtypes and the evolution of precision medicine in colorectal cancer. Nat Rev Cancer 17:79–92

    CAS  PubMed  Google Scholar 

  18. El Messaoudi S, Rolet F, Mouliere F et al (2013) Circulating cell free DNA: preanalytical considerations. Clin Chim Acta 424:222–230

    CAS  PubMed  Google Scholar 

  19. Fearon ER, Vogelstein B (1990) A genetic model for colorectal tumorigenesis. Cell 61:759–767

    CAS  PubMed  Google Scholar 

  20. Frattini M, Gallino G, Signoroni S et al (2008) Quantitative and qualitative characterization of plasma DNA identifies primary and recurrent colorectal cancer. Cancer Lett 263:170–181

    CAS  PubMed  Google Scholar 

  21. Gerlinger M, Rowan AJ, Horswell S et al (2012) Intratumor heterogeneity and branched evolution revealed by multiregion sequencing. N Engl J Med 366:883–892

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Grasselli J, Elez E, Caratu G et al (2017) Concordance of blood- and tumor-based detection of RAS mutations to guide anti-EGFR therapy in metastatic colorectal cancer. Ann Oncol 28:1294–1301

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Grutzmann R, Molnar B, Pilarsky C et al (2008) Sensitive detection of colorectal cancer in peripheral blood by septin 9 DNA methylation assay. PLoS One 3:e3759

    PubMed  PubMed Central  Google Scholar 

  24. Heitzer E, Haque IS, Roberts CES et al (2019) Current and future perspectives of liquid biopsies in genomics-driven oncology. Nat Rev Genet 20:71–88

    CAS  PubMed  Google Scholar 

  25. Khan KH, Cunningham D, Werner B et al (2018) Longitudinal liquid biopsy and mathematical modeling of clonal evolution forecast time to treatment failure in the PROSPECT‑C phase II colorectal cancer clinical trial. Cancer Discov 8:1270–1285

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Kim K, Shin DG, Park MK et al (2014) Circulating cell-free DNA as a promising biomarker in patients with gastric cancer: diagnostic validity and significant reduction of cfDNA after surgical resection. Ann Surg Treat Res 86:136–142

    PubMed  PubMed Central  Google Scholar 

  27. Leon SA, Shapiro B, Sklaroff DM et al (1977) Free DNA in the serum of cancer patients and the effect of therapy. Cancer Res 37:646–650

    CAS  PubMed  Google Scholar 

  28. Lofton-Day C, Model F, Devos T et al (2008) DNA methylation biomarkers for blood-based colorectal cancer screening. Clin Chem 54:414–423

    CAS  PubMed  Google Scholar 

  29. Mandel P, Metais P (1948) Les acides nucléiques du plasma sanguin chez l’homme. C R Seances Soc Biol Fil 142:241–243

    CAS  PubMed  Google Scholar 

  30. Misale S, Yaeger R, Hobor S et al (2012) Emergence of KRAS mutations and acquired resistance to anti-EGFR therapy in colorectal cancer. Nature 486:532–536

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Mouliere F, Chandrananda D, Piskorz AM et al (2018) Enhanced detection of circulating tumor DNA by fragment size analysis. Sci Transl Med 10(466):eaat4921. https://doi.org/10.1126/scitranslmed.aat4921

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Mouliere F, El Messaoudi S, Pang D et al (2014) Multi-marker analysis of circulating cell-free DNA toward personalized medicine for colorectal cancer. Mol Oncol 8:927–941

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Murtaza M, Dawson SJ, Pogrebniak K et al (2015) Multifocal clonal evolution characterized using circulating tumour DNA in a case of metastatic breast cancer. Nat Commun 6:8760

    PubMed  PubMed Central  Google Scholar 

  34. Normanno N, Esposito Abate R, Lambiase M et al (2018) RAS testing of liquid biopsy correlates with the outcome of metastatic colorectal cancer patients treated with first-line FOLFIRI plus cetuximab in the CAPRI-GOIM trial. Ann Oncol 29:112–118

    CAS  PubMed  Google Scholar 

  35. Overman MJ, Mcdermott R, Leach JL et al (2017) Nivolumab in patients with metastatic DNA mismatch repair-deficient or microsatellite instability-high colorectal cancer (CheckMate 142): an open-label, multicentre, phase 2 study. Lancet Oncol 18:1182–1191

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Pantel K, Alix-Panabieres C (2019) Liquid biopsy and minimal residual disease—latest advances and implications for cure. Nat Rev Clin Oncol 16:409–424

    CAS  PubMed  Google Scholar 

  37. Russo M, Misale S, Wei G et al (2016) Acquired resistance to the TRK inhibitor entrectinib in colorectal cancer. Cancer Discov 6:36–44

    CAS  PubMed  Google Scholar 

  38. Siravegna G, Bardelli A (2014) Genotyping cell-free tumor DNA in the blood to detect residual disease and drug resistance. Genome Biol 15:449

    PubMed  PubMed Central  Google Scholar 

  39. Siravegna G, Marsoni S, Siena S et al (2017) Integrating liquid biopsies into the management of cancer. Nat Rev Clin Oncol 14:531–548

    CAS  PubMed  Google Scholar 

  40. Siravegna G, Mussolin B, Buscarino M et al (2015) Clonal evolution and resistance to EGFR blockade in the blood of colorectal cancer patients. Nat Med 21:795–801

    CAS  PubMed  PubMed Central  Google Scholar 

  41. Sorenson GD, Pribish DM, Valone FH et al (1994) Soluble normal and mutated DNA sequences from single-copy genes in human blood. Cancer Epidemiol Biomarkers Prev 3:67–71

    CAS  PubMed  Google Scholar 

  42. Sottoriva A, Kang H, Ma Z et al (2015) A big bang model of human colorectal tumor growth. Nat Genet 47:209–216

    CAS  PubMed  PubMed Central  Google Scholar 

  43. Stroun M, Anker P, Maurice P et al (1989) Neoplastic characteristics of the DNA found in the plasma of cancer patients. Oncology 46:318–322

    CAS  PubMed  Google Scholar 

  44. Thierry AR, Mouliere F, El Messaoudi S et al (2014) Clinical validation of the detection of KRAS and BRAF mutations from circulating tumor DNA. Nat Med 20:430–435

    CAS  PubMed  Google Scholar 

  45. Tie J, Kinde I, Wang Y et al (2015) Circulating tumor DNA as an early marker of therapeutic response in patients with metastatic colorectal cancer. Ann Oncol 26:1715–1722

    CAS  PubMed  PubMed Central  Google Scholar 

  46. Tie J, Wang Y, Tomasetti C et al (2016) Circulating tumor DNA analysis detects minimal residual disease and predicts recurrence in patients with stage II colon cancer. Sci Transl Med 8:346ra392

    Google Scholar 

  47. Vogelstein B, Kinzler KW (1999) Digital PCR. Proc Natl Acad Sci Usa 96:9236–9241

    CAS  PubMed  Google Scholar 

  48. Wan JCM, Massie C, Garcia-Corbacho J et al (2017) Liquid biopsies come of age: towards implementation of circulating tumour DNA. Nat Rev Cancer 17:223–238

    CAS  PubMed  Google Scholar 

  49. Wang JY, Hsieh JS, Chang MY et al (2004) Molecular detection of APC, K‑ras, and p53 mutations in the serum of colorectal cancer patients as circulating biomarkers. World J Surg 28:721–726

    PubMed  Google Scholar 

  50. Zheng Z, Liebers M, Zhelyazkova B et al (2014) Anchored multiplex PCR for targeted next-generation sequencing. Nat Med 20:1479–1484

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Pathologie, Universitätsmedizin Mainz, Langenbeckstr. 1, 55131, Mainz, Deutschland

    A. Haupts, W. Roth & N. Hartmann

Authors
  1. A. Haupts
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. W. Roth
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. N. Hartmann
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A. Haupts.

Ethics declarations

Interessenkonflikt

A. Haupts, W. Roth und N. Hartmann geben an, dass kein Interessenkonflikt besteht.

Für diesen Beitrag wurden von den Autoren keine Studien an Menschen oder Tieren durchgeführt. Für die aufgeführten Studien gelten die jeweils dort angegebenen ethischen Richtlinien.

The supplement containing this article is not sponsored by industry.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Haupts, A., Roth, W. & Hartmann, N. Liquid Biopsy im kolorektalen Karzinom. Pathologe 40 (Suppl 3), 244–251 (2019). https://doi.org/10.1007/s00292-019-00698-3

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00292-019-00698-3

Schlüsselwörter

Keywords

Search

Navigation