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Detection of Circulating Tumor DNA in the Blood of Cancer Patients: An Important Tool in Cancer Chemoprevention

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Book cover Cancer Chemoprevention

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1379))

Abstract

Liquid biopsies represent novel promising tools to determine the impact of clonal heterogeneity on clinical outcomes with the potential to identify novel therapeutic targets in cancer patients. We developed a low-coverage whole-genome sequencing approach in order to noninvasively establish copy number aberrations in plasma DNA from metastasized cancer patients. Using plasma-Seq we were able to monitor genetic evolution including the acquirement of novel copy number changes, such as focal amplifications and chromosomal polysomies. The big advantage of our approach is that it can be performed on a benchtop sequencer, speed, and cost-effectiveness. Therefore, plasma-Seq represents an easy, fast, and affordable tool to provide the urgently needed genetic follow-up data. Here we describe our method including plasma DNA extraction, library preparation, and bioinformatic analyses.

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References

  1. Jung M, Klotzek S, Lewandowski M et al (2003) Changes in concentration of DNA in serum and plasma during storage of blood samples. Clin Chem 49:1028–1029

    Article  CAS  PubMed  Google Scholar 

  2. Lui YY, Chik KW, Chiu RW et al (2002) Predominant hematopoietic origin of cell-free DNA in plasma and serum after sex-mismatched bone marrow transplantation. Clin Chem 48:421–427

    CAS  PubMed  Google Scholar 

  3. Bettegowda C, Sausen M, Leary RJ et al (2014) Detection of circulating tumor DNA in early- and late-stage human malignancies. Sci Transl Med 6:22ra424

    Article  Google Scholar 

  4. Chan KC, Jiang P, Chan CW et al (2013) Noninvasive detection of cancer-associated genome-wide hypomethylation and copy number aberrations by plasma DNA bisulfite sequencing. Proc Natl Acad Sci U S A 110:18761–18768

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Chan KC, Jiang P, Zheng YW et al (2013) Cancer genome scanning in plasma: detection of tumor-associated copy number aberrations, single-nucleotide variants, and tumoral heterogeneity by massively parallel sequencing. Clin Chem 59:211–224

    Article  CAS  PubMed  Google Scholar 

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

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Forshew T (2012) Noninvasive identification and monitoring of cancer mutations by targeted deep sequencing of plasma DNA. Sci Transl Med 4:136ra68

    Article  PubMed  Google Scholar 

  8. Heidary M, Auer M, Ulz P et al (2014) The dynamic range of circulating tumor DNA in metastatic breast cancer. Breast Cancer Res 16:421

    Article  PubMed Central  PubMed  Google Scholar 

  9. Heitzer E, Auer M, Hoffmann EM et al (2013) Establishment of tumor-specific copy number alterations from plasma DNA of patients with cancer. Int J Cancer 133:346–356

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  10. Heitzer E, Auer M, Ulz P et al (2013) Circulating tumor cells and DNA as liquid biopsies. Genome Med 5:73

    Article  PubMed Central  PubMed  Google Scholar 

  11. Heitzer E, Ulz P, Belic J et al (2013) Tumor-associated copy number changes in the circulation of patients with prostate cancer identified through whole-genome sequencing. Genome Med 5:30

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  12. Heitzer E, Ulz P, Geigl JB (2014) Circulating tumor DNA as a liquid biopsy for cancer. Clin Chem 61:112

    Article  PubMed  Google Scholar 

  13. Mohan S, Heitzer E, Ulz P et al (2014) Changes in colorectal carcinoma genomes under anti-egfr therapy identified by whole-genome plasma DNA sequencing. PLoS Genet 10:e1004271

    Article  PubMed Central  PubMed  Google Scholar 

  14. Murtaza M (2013) Non-invasive analysis of acquired resistance to cancer therapy by sequencing of plasma DNA. Nature 497:108–112

    Article  CAS  PubMed  Google Scholar 

  15. Baslan T, Kendall J, Rodgers L et al (2012) Genome-wide copy number analysis of single cells. Nat Protoc 7:1024–1041

    Article  CAS  PubMed  Google Scholar 

  16. Li H, Durbin R (2009) Fast and accurate short read alignment with burrows-wheeler transform. Bioinformatics 25:1754–1760

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Li H, Handsaker B, Wysoker A et al (2009) The sequence alignment/map format and samtools. Bioinformatics 25:2078–2079

    Article  PubMed Central  PubMed  Google Scholar 

  18. R:. R: A language and environment for statistical computing. http://www.R-project.org

  19. Lai W, Choudhary V, Park PJ (2008) Cghweb: a tool for comparing DNA copy number segmentations from multiple algorithms. Bioinformatics 24:1014–1015

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  20. Navin N, Kendall J, Troge J et al (2011) Tumour evolution inferred by single-cell sequencing. Nature 472:90–94

    Article  PubMed Central  CAS  PubMed  Google Scholar 

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Correspondence to Ellen Heitzer .

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Ulz, P., Auer, M., Heitzer, E. (2016). Detection of Circulating Tumor DNA in the Blood of Cancer Patients: An Important Tool in Cancer Chemoprevention. In: Strano, S. (eds) Cancer Chemoprevention. Methods in Molecular Biology, vol 1379. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3191-0_5

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  • DOI: https://doi.org/10.1007/978-1-4939-3191-0_5

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3190-3

  • Online ISBN: 978-1-4939-3191-0

  • eBook Packages: Springer Protocols

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