Next-Generation Genotyping by Digital PCR to Detect and Quantify the BRAF V600E Mutation in Melanoma Biopsies.

The detection of the BRAF V600E mutation in melanoma samples is used to select patients who should respond to BRAF inhibitors. Different techniques are routinely used to determine BRAF status in clinical samples. However, low tumor cellularity and tumor heterogeneity can affect the sensitivity of somatic mutation detection. Digital PCR (dPCR) is a next-generation genotyping method that clonally amplifies nucleic acids and allows the detection and quantification of rare mutations. Our aim was to evaluate the clinical routine performance of a new dPCR-based test to detect and quantify BRAF mutation load in 47 paraffin-embedded cutaneous melanoma biopsies. We compared the results obtained by dPCR with high-resolution melting curve analysis and pyrosequencing or with one of the allele-specific PCR methods available on the market. dPCR showed the lowest limit of detection. dPCR and allele-specific amplification detected the highest number of mutated samples. For the BRAF mutation load quantification both dPCR and pyrosequencing gave similar results with strong disparities in allele frequencies in the 47 tumor samples under study (from 0.7% to 79% of BRAF V600E mutations/sample). In conclusion, the four methods showed a high degree of concordance. dPCR was the more-sensitive method to reliably and easily detect mutations. Both pyrosequencing and dPCR could quantify the mutation load in heterogeneous tumor samples.

Prognostic impact of the inclusion of uPA/PAI-1 tumor levels in the current adjuvant treatment decision-making for early breast cancer.

AIMS: Following the introduction of new adjuvant therapies we wanted to reappraise the prognostic and predictive value of uPA/PAI-1 in early breast cancer. PATIENTS & METHODS: This monocentric retrospective study included 652 patients who had curative surgery between 2006 and 2011 and adjuvant treatment decision-making, taking into account uPA/PAI-1 tumor levels. RESULTS: uPA and PAI-1 levels were associated with classical clinicopathological parameters and adjuvant chemotherapy decision, but not with peritumoral vascular invasion (PVI; also known as peritumoral vascular emboli). HER2 overexpression, PVI and uPA/PAI-1 levels were not significantly associated with relapse-free survival in univariate analysis. In multivariate analysis, T stage, N stage and progesterone receptors were the only independent relapse-free survival predictive factors. CONCLUSION: The absence of an association between uPA/PAI-1 and PVI allows their concomitant consideration in adjuvant treatment discussion. The overall good prognosis of patients with high uPA/PAI-1 levels might be linked to the uPA/PAI-1 predictive value and the inclusion of these parameters in adjuvant guidelines.

[Evaluation of (-2)proPSA in combination with total PSA and free PSA for the early detection of prostate cancer]. / Évaluation du test (-2)proPSA combiné au PSA total et PSA libre pour la détection précoce du cancer de la prostate.

The analytical and clinical validation of new biomarkers for the early detection of prostate is necessary. (-2)proPSA, total PSA and free PSA values are used to calculate a standardized PHI index linked to a higher probability of a positive biopsy in patients with PSA levels between 3-4 and 10 ng/L, the gray zone for prostate cancer diagnosis. The purpose of this study is to validate the analytical performance of the (-2)proPSA and to determine the predictive value of PHI for the early detection of prostate cancer. Analytical performances are correct. It is not necessary to dilute samples before analysis. The stability of (-2)proPSA is good until at least 3 hours at room temperature before centrifugation. The study of the PSAT, PSAL, (-2)proPSA and PHI values in a population of patients consulting for an early prostate cancer diagnosis shows that the index PHI is the most powerful predictive marker of cancer with an area under ROC curve of 0.70, whereas it is only 0.56 for total PSA.

Anemia and iron biomarkers in patients with early breast cancer. Diagnostic value of hepcidin and soluble transferrin receptor quantification.

BACKGROUND: Anemia, a frequent and deleterious condition in patients with cancer, is mainly caused by chemotherapy toxicity, iron deficiency, or inflammation. We evaluated the baseline iron metabolism biomarkers and their association with anemia occurrence during chemotherapy in patients with early breast cancer (EBC). METHODS: In this monocentric retrospective study, classical iron metabolism markers and new biomarkers as well as sTfR and hepcidin were assessed at baseline in 347 patients with EBC who received a sequential taxane and anthracycline-based regimen between April 2007 and October 2009. Hemoglobin level was measured every 21 days. RESULTS: Thirty-five patients had baseline iron deficiency and 13 inflammatory iron sequestration. In multivariate analysis, only high sTfR (OR=27.6, p<0.001, 95% CI 8.74-87) and pre-menopausal status (OR=7.3, 95% CI 0.04-0.43, p=0.001) remained statistically associated with iron deficiency. High hepcidin values and inflammatory iron sequestration were significantly associated (p=0.032). In total 6.1% patients had baseline anemia and 86.2% patients developed anemia during chemotherapy (41 had grade ≥2 anemia). Baseline hemoglobin below 13 g/dL and low hepcidin levels were the two independent predictive factors of severe anemia. CONCLUSIONS: In early breast cancer treated by chemotherapy, only baseline hemoglobin and hepcidin levels are independent predictive factors of anemic syndrome occurrence.

[Iron metabolism in breast cancer: knowledge and future]. / Métabolisme du fer et cancer du sein: connaissances et perspectives.

Iron plays a fundamental role in biology and its concentration in living organisms is regulated very precisely. Many molecules of storage and transportation are used to maintain the intracellular homeostasis. Cancer cells have alterations in this balance. Recent studies have shown that breast cancer cells present abnormal expression of several proteins such as hepcidin and ferroportin. A prognostic impact of these alterations has been reported in patients with breast cancer. Regulatory molecules of iron metabolism could become therapeutic targets. This is an innovative approach that has emerged for treating a cancer which, despite advances in treatment and the emergence of targeted therapies, remains the leading cause of cancer death in women.

[HE4, a novel marker for epithelial ovarian cancer: evaluation of analytical performances]. / HE4, un nouveau marqueur des cancers épithéliaux ovariens: évaluation des performances analytiques.

Human epididymis protein 4 (HE4) is a novel marker for ovarian cancer. HE4 exhibits a high sensitivity to detect ovarian cancer and can be used with CA125 as a predictor of malignancy. Additional uses of HE4 are as an aid of monitoring response to therapy for patients with invasive ovarian cancer and as a marker to detect recurrences in the follow-up after treatment of the primary tumor. The HE4 EIA, an enzyme immunoassay for the quantitative determination of HE4 in human serum developed by Fujirebio Diagnostic Inc. (Tokyo, Japan), is now available with a CE-IVD label in Europe (Diasource, Nivelles, Belgium). The aim of the study was to evaluate according to the COFRAC LAB GTA 04 guide, the analytical performance of the test, using 4 standardized samples (target values: 49.8, 140.4, 167.6 and 415.2 pmol/L) and serum samples from patients with ovarian cancer treated in our institution. Intra- and inter-assay precisions showed coefficients of variation less than 10%. The low limit of detection (4 pmol/L) and the limit of quantitation (8 pmol/L) are suitable for clinical samples assessment. The assay mean dilution linearity is 100 +/- 10% (90 to 107% of recovery). Globally, the uncertainty varied from 13.1% (low values) to 28.1% (elevated values). We conclude that the HE4 EIA from Fujirebio Diagnostic Inc. displayed convenient analytical performances that allows its use it clinical practice.