Dynamic sequencing of circulating tumor DNA: novel noninvasive cancer biomarker

Circulating tumor DNA (ctDNA) in fluids has gained attention because ctDNA seems to identify tumor-specific abnormalities, which could be used for diagnosis, follow-up of treatment, and prognosis: the so-called liquid biopsy. Liquid biopsy is a minimally invasive approach and presents the sum of ctDNA from primary and secondary tumor sites. It has been possible not only to quantify the amount of ctDNA but also to identify (epi)genetic changes. Specific mutations in genes have been identified in the plasma of patients with several types of cancer, which highlights ctDNA as a possible cancer biomarker. However, achieving detectable concentrations of ctDNA in body fluids is not an easy task. ctDNA fragments present a short half-life, and there are no cut-off values to discriminate high and low ctDNA concentrations. Here, we discuss the use of ctDNA as a cancer biomarker, the main methodologies, the inherent difficulties, and the clinical predictive value of ctDNA.

Download Full-text

Circulating tumor DNA as a cancer biomarker

Journal of Clinical Oncology ◽

10.1200/jco.2008.26.15_suppl.4122 ◽

2008 ◽

Vol 26 (15_suppl) ◽

pp. 4122-4122

Author(s):

F. Diehl ◽

K. Schmidt ◽

M. A. Choti ◽

S. Goodman ◽

K. Romans ◽

...

Keyword(s):

Circulating Tumor Dna ◽

Cancer Biomarker ◽

Tumor Dna

Download Full-text

Measurement of circulating tumor DNA as a cancer biomarker in gastrointestinal malignancies using a novel next-generation sequencing method.

Journal of Clinical Oncology ◽

10.1200/jco.2014.32.3_suppl.217 ◽

2014 ◽

Vol 32 (3_suppl) ◽

pp. 217-217

Author(s):

Edward Samuel James ◽

Azeet Narayan ◽

Stacey Stein ◽

Jill Lacy ◽

Abhijit Patel ◽

...

Keyword(s):

Locally Advanced ◽

Circulating Tumor Dna ◽

Cancer Biomarker ◽

Multiple Time ◽

Metastatic Colon Cancer ◽

Next Generation ◽

Gastrointestinal Malignancies ◽

Time Points ◽

Multiple Time Points ◽

Tumor Dna

217 Background: Circulating tumor DNA (ctDNA) holds promise as a highly specific cancer biomarker. The presence of mutant tumor-derived DNA fragments in the blood provides an opportunity to non-invasively assess tumor mutation profiles and to quantify changes in tumor DNA levels over time. Methods: After obtaining informed consent, plasma samples were collected prospectively at multiple time points in a cohort of patients (pts) with various gastrointestinal (GI) malignancies in the locally advanced, metastatic and adjuvant settings. Hotspot regions of genes known to be commonly mutated in GI tumors were amplified by multiplexed PCR, and the resultant amplicons were subjected to next-generation ultra-deep sequencing. Suppression of sequencer and PCR errors allowed mutations to be identified and quantified with a sensitivity of approximately 1 variant in 5,000 molecules. Sample-specific barcoding allowed simultaneous analysis of up to 96 samples. Results: 29 out of 74 available samples from 17 pts were analyzed for the presence of ctDNA. 3 pts had KRAS mutations exclusively. 2 pts had mutations in two of the evaluated genes: KRAS plus PIK3CA and KRAS plus TP53. 2 pts with measurable mutant ctDNA had samples analyzed at multiple time points. One of these patients with locally advanced pancreatic cancer initially had a KRAS G12D mutation which disappeared after treatment with chemo-radiation therapy, but then he developed significantly rising levels of a different KRAS G12L mutation just preceding the diagnosis of liver metastases by imaging. A second pt with metastatic colon cancer had high levels of ctDNA prior to treatment that decreased dramatically after initiation of mFOLFOX and bevacizumab therapy. Conclusions: Our initial analysis indicates that our technique is able to quantify multiple mutations at very low copy numbers accurately in this cohort of pts with GI cancers. Moreover, we were able to monitor changes in mutation type and quantity using this non-invasive “liquid biopsy” approach. Analysis of remaining samples is ongoing, and additional longitudinal data will be presented.

Download Full-text