Prognostic and predictive value of circulating tumor cell analysis in colorectal cancer patients.

OBJECTIVE: The aim of this study was to assess the prognostic and predictive values of circulating tumor cell (CTC) analysis in colorectal cancer patients. PATIENTS AND METHODS: Presence of CTCs was evaluated in 60 colorectal cancer patients before systemic therapy--from which 33 patients were also evaluable for CTC analysis during the first 3 months of treatment--through immunomagnetic enrichment, using the antibodies BM7 and VU1D9 (targeting mucin 1 and EpCAM, respectively), followed by real-time RT-PCR analysis of the tumor-associated genes KRT19, MUC1, EPCAM, CEACAM5 and BIRC5. RESULTS: Patients were stratified into groups according to CTC detection (CTC negative, when all marker genes were negative; and CTC positive when at least one of the marker genes was positive). Patients with CTC positivity at baseline had a significant shorter median progression-free survival (median PFS 181.0 days; 95% CI 146.9-215.1) compared with patients with no CTCs (median PFS 329.0 days; 95% CI 299.6-358.4; Log-rank P < .0001). Moreover, a statistically significant correlation was also founded between CTC detection during treatment and radiographic findings at the 6 month staging. This correlation applied to CTC results before therapy (odds ratio (OR), 6.22), 1 to 4 weeks after beginning of treatment (OR, 5.50), 5 to 8 weeks after beginning of treatment (OR, 7.94) 9 to 12 weeks after beginning of treatment (OR, 14.00) and overall CTC fluctuation during the course of treatment (OR, 20.57). CONCLUSION: The present study provides evidence of a strong correlation between CTC detection and radiographic disease progression in patients receiving chemotherapy for colorectal cancer. Our results suggest that in addition to the current prognostic factors, CTC analysis represent a potential complementary tool for prediction of colorectal cancer patients' outcome. Moreover, the present test allows for molecular characterization of CTCs, which may be of relevance to the creation of personalized therapies.

Development of a molecular multimarker assay for the analysis of circulating tumor cells in adenocarcinoma patients.

BACKGROUND: The analysis of circulating tumor cells (CTCs) is emerging as a promising diagnostic tool in oncology. However, even if a variety of methods for CTC isolation have been already developed, their specificity and/or sensitivity still remain problematic. The aim of this study was to develop an immunomagnetic/real-time reverse transcription polymerase chain reaction (RT-PCR) assay for the molecular detection of circulating tumor cells (CTCs) in peripheral blood (PB) of adenocarcinoma cancer patients. METHODS: The presence of CTCs was evaluated in 945 PB blood samples from 247 adenocarcinoma cancer patients and in 42 healthy controls by immunomagnetic enrichment using the antibodies BM7 and VU1D9 followed by real-time RT-PCR analysis of the marker genes KRT19, MUC1, EPCAM, CEACAM5, BIRCS, SCGB2A2, and ERBB2. RESULTS: The developed assay showed not only high specificity, as none of the healthy controls were found positive for the multimarker gene panel, but also great sensitivity as CTCs were detected in adenocarcinomas arising from 10 different organs. According to tumor primary origin, CTC positivity was detected in 33.3% of Ampulla of Vater adenocarcinomas, 69.6% of bile ducts adenocarcinomas, 61.3% of breast adenocarcinomas, 61.3% of cardia adenocarcinomas, 60.6% of colon adenocarcinomas, 66.7% of esophagus adenocarcinomas, 57.1% of pancreas adenocarcinomas, 66.7% of rectum adenocarcinomas, 33.3% of small intestine adenocarcinomas, and 62.2% of stomach adenocarcinomas. CONCLUSIONS: Our results suggest that the current developed technique can be used to detect CTCs in all major adenocarcinomas, with great sensitivity without compromising specificity.

Multimarker gene analysis of circulating tumor cells in pancreatic cancer patients: a feasibility study.

OBJECTIVE: The aim of this study was to develop an immunomagnetic/real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay and assess its clinical value for the molecular detection of circulating tumor cells (CTCs) in peripheral blood of pancreatic cancer patients. METHODS: The presence of CTCs was evaluated in 34 pancreatic cancer patients before systemic therapy and in 40 healthy controls, through immunomagnetic enrichment, using the antibodies BM7 and VU1D9 [targeting mucin 1 and epithelial cell adhesion molecule (EpCAM), respectively], followed by real-time RT-PCR analysis of the genes KRT19, MUC1, EPCAM, CEACAM5 and BIRC5. RESULTS: The developed assay showed high specificity, as none of the healthy controls were found to be positive for the multimarker gene panel. CTCs were detected in 47.1% of the pancreatic cancer patients before the beginning of systemic treatment. Shorter median progression-free survival (PFS) was observed for patients who had at least one detectable tumor-associated transcript, compared with patients who were CTC negative. Median PFS time was 66.0 days [95% confidence interval (CI) 44.8-87.2] for patients with baseline CTC positivity and 138.0 days (95% CI 124.1-151.9) for CTC-negative patients (p = 0.01, log-rank test). CONCLUSION: Our results suggest that in addition to the current prognostic methods, CTC analysis represents a potential complementary tool for prediction of outcome in pancreatic cancer patients.

Multicenter evaluation of a rapid electrochemiluminescent adrenocorticotropic hormone (ACTH) immunoassay.

BACKGROUND: Measuring plasma adrenocorticotropic hormone (ACTH) is a key step in the differential diagnosis of hypothalamic-pituitary-adrenal disorders. METHODS: The recently developed electrochemiluminescence Elecsys ACTH immunoassay (Roche Diagnostics, Mannheim, Germany) was evaluated at six clinical laboratories on the Modular E170 and/or the Elecsys 2010 (Roche Diagnostics) immunoanalysers. RESULTS: The within-run and between-run imprecision was