Performance of standardized BEAMing platform for detecting RAS mutations in the blood of metastatic colorectal cancer (mCRC) patients.

Jones, Frederick S.; Edelstein, Dan; Wichner, Katharina; Ross, C. M. D.; Stieler, Kristin; Boehm, Verena; Lenfert, Eva; Bien, Jessica; Lukas, Antje; Macioszek, Jerzy

doi:10.1200/jco.2016.34.15_suppl.11538

Cited by 7 publications

(9 citation statements)

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“…Plasma was prepared either after 2 h or after 3 d of storage at RT, and samples were subsequently tested for 33 different mutations in the KRAS and NRAS genes using the OncoBEAM RAS panel, a standardized BEAMing approach [28]. …”

Section: Resultsmentioning

confidence: 99%

“…We expect that theses tubes can be applied for any cancer type and liquid biopsy application using BEAMing and Plasma Safe-Sequencing (Safe-SeqS). Indeed, the CE-IVD version of the cfDNA BCTs is now recommended as the preferred collection device for the CE-IVD OncoBEAM RAS CRC kit, a liquid biopsy test for RAS mutation profiling in colorectal cancer [28]. Even if these tubes have some limitations with respect to the temperature ranges as well as biomarker compatibility, liquid biopsy testing can now be performed for patients who previously missed easy access to this type of molecular mutation profiling.…”

Section: Discussionmentioning

confidence: 99%

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Performance of Streck cfDNA Blood Collection Tubes for Liquid Biopsy Testing

et al. 2016

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ObjectivesMaking liquid biopsy testing widely available requires a concept to ship whole blood at ambient temperatures while retaining the integrity of the cell-free DNA (cfDNA) population and stability of blood cells to prevent dilution of circulating tumor DNA (ctDNA) with wild-type genomic DNA. The cell- and DNA-stabilizing properties of Streck Cell-Free DNA BCT blood collection tubes (cfDNA BCTs) were evaluated to determine if they can be utilized in combination with highly sensitive mutation detection technologies.MethodsVenous blood from healthy donors or patients with advanced colorectal cancer (CRC) was collected in cfDNA BCTs and standard K2EDTA tubes. Tubes were stored at different temperatures for various times before plasma preparation and DNA extraction. The isolated cfDNA was analyzed for overall DNA yield of short and long DNA fragments using qPCR as well as for mutational changes using BEAMing and Plasma Safe-Sequencing (Safe-SeqS).ResultsCollection of whole blood from healthy individuals in cfDNA BCTs and storage for up to 5 days at room temperature did not affect the DNA yield and mutation background levels (n = 60). Low-frequency mutant DNA spiked into normal blood samples as well as mutant circulating tumor DNA in blood samples from CRC patients collected in cfDNA BCTs were reliably detected after 3 days of storage at room temperature. However, blood samples stored at ≤ 10°C and at 40°C for an extended period of time showed elevated normal genomic DNA levels and an abnormally large cellular plasma interface as well as lower plasma volumes.ConclusionWhole blood shipped in cfDNA BCTs over several days can be used for downstream liquid biopsy testing using BEAMing and Safe-SeqS. Since the shipping temperature is a critical factor, special care has to be taken to maintain a defined room temperature range to obtain reliable mutation testing results.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Performance of Streck cfDNA Blood Collection Tubes for Liquid Biopsy Testing

et al. 2016

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“…After the initial response, the disease was progressive. The patient was in good general health (ECOG 0) and refused further biopsy; therefore, a liquid biopsy as previously reported was performed [1, 2]. No RAS mutations were detected and a second-line therapy including cetuximab was initiated.…”

Section: Introductionmentioning

confidence: 95%

“…Recently, liquid biopsy, a noninvasive blood-based RAS mutation analysis of cell-free circulating tumor DNA (ctDNA), has been reported as a suitable method for mutational analysis in patients with mCRC [1, 2]. In contrast to tissue based RAS status determination, liquid biopsy is able to address tumor heterogeneity and treatment-related dynamic changes of molecular profiles [3–6].…”

Section: Introductionmentioning

confidence: 99%

“…After the deep anterior rectum resection, a systemic chemotherapy combined with cetuximab was initiated, and after confirmation of a partial response this treatment was continued for 33 cycles. During this period, no RAS mutation was detectable by liquid biopsy (Figure 2) [1, 2]. However, after the 33rd cycle (July 2015) of treatment with continuous application of cetuximab, low frequency KRAS exon 3 mutations were detectable by liquid biopsy in the plasma of the patient for the first time.…”

Section: Introductionmentioning

confidence: 99%

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Clinical Application of Liquid Biopsy in Targeted Therapy of Metastatic Colorectal Cancer

Trojan

Klein-Scory

Koch

et al. 2017

Case Reports in Oncological Medicine

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Background. Colorectal cancers (CRC) shed DNA into blood circulation. There is growing evidence that the analysis of circulating tumor DNA can be effectively used for monitoring of disease, to track tumor heterogeneity and to evaluate response to treatment. Case Presentation. Here, we describe two cases of patients with advanced CRC. The first case is about a patient with no available tissue for analysis of RAS mutation status. Liquid biopsy revealed RAS-wild-type and the therapy with anti-EGFR (epidermal growth factor receptor) monoclonal antibody cetuximab could be initiated. In the second case, the mutational profile of a patient with initial wild-type RAS-status was continually tracked during the course of treatment. An acquired KRAS exon 3 mutation was detected. The number of KRAS mutated fragments decreased continuously after the discontinuation of the therapy with EGFR-specific antibodies. Conclusion. Liquid biopsy provides a rapid genotype result, which accurately reproduces the current mutation status of tumor tissue. Furthermore, liquid biopsy enables close monitoring of the onset of secondary resistance to anti-EGFR therapy.

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