A Novel Detection Method of Breast Cancer through a Simple Panel of Biomarkers.

Circulating tumor cells (CTCs) have been identified as responsible for the spread of tumors to other organs of the body. In this sense, the development of sensitive and specific assays for their detection is important to reduce the number of deaths due to metastases. Here, we assessed whether the detection of CTCs in peripheral blood can serve in the construction of a panel of diagnosis and monitoring treatments of breast cancer (BC), focusing on the expression of markers of epithelial-mesenchymal transition. Through analyzing the blood from women without breast alterations (control), women with benign alterations, women with breast cancer without chemotherapy, and women with breast cancer with chemotherapy, we identified the best markers by transcriptional levels and determined three profiles of CTCs (mesenchymal, intermediate, and epithelial) by flow cytometry which, combined, can be used for diagnosis and therapy monitoring with sensitivity and specificity between 80% and 100%. Therefore, we have developed a method for detecting breast cancer based on the analysis of CTC profiles by epithelial-mesenchymal transition markers which, combined, can be used for the diagnosis and monitoring of therapy.

Altered Leukocyte Sphingolipid Pathway in Breast Cancer.

Sphingolipid metabolism pathway is essential in membrane homeostasis, and its dysfunction has been associated with favorable tumor microenvironment, disease progression, and chemotherapy resistance. Its major components have key functions on survival and proliferation, with opposing effects. We have profiled the components of the sphingolipid pathway on leukocytes of breast cancer (BC) patients undergoing chemotherapy treatment and without, including the five sphingosine 1-phosphate (S1P) receptors, the major functional genes, and cytokines, in order to better understand the S1P signaling in the immune cells of these patients. To the best of our knowledge, this is the first characterization of the sphingolipid pathway in whole blood of BC patients. Skewed gene profiles favoring high SPHK1 expression toward S1P production during BC development was observed, which was reversed by chemotherapy treatment, and reached similar levels to those found in healthy donors. Such levels were also correlated with high levels of TNF-α. Our data revealed an important role of the sphingolipid pathway in immune cells in BC with skewed signaling of S1P receptors, which favored cancer development even under chemotherapy, and may probably be a trigger of cancer resistance. Thus, these molecules must be considered as a target pathway for combined BC therapeutics.

A Lower Serum Antioxidant Capacity as a Distinctive Feature for Women with HER2+ Breast Cancer: A Preliminary Study.

The overexpression of HER2 in breast cancer (BC) can contribute to redox imbalance, which is related to damage and structural modification in many biomolecules. To the best of our knowledge, this is the first study that has investigated the infrared spectrum wavenumbers obtained by ATR-FTIR and their relationship with the levels of redox status markers such as reduced glutathione, superoxide dismutase (SOD), catalase, Ferric Reducing Antioxidant Power (FRAP), and protein carbonyl among women with HER2+ BC, HER2- BC, and benign breast disease (BBD). The study was conducted with 25 women, 17 of whom were diagnosed with BC (6 HER2+ and 11 HER2-) and 8 with BBD. Our results indicate HER2+ BC cases could be distinguished from HER2- BC and BBD cases by their serum's antioxidant capacity [HER2+ BC vs. HER2- BC (AUC = 0.818; specificity = 81.82%; sensitivity = 66.67%); HER2+ BC vs. BBD (AUC = 0.875; specificity = 75%; sensitivity = 83.33%)]. The changes in biochemical terms that occur in serum as a result of the scarcity of antioxidants are related to a peculiar fingerprint in the infrared spectrum obtained by ATR-FTIR. In the serum of women with BBD, the SOD enzyme level is the highest, and this characteristic allowed us to distinguish them from HER2- BC. Finally, data regarding the serological antioxidant capacity of FRAP and the infrared spectrum by ATR-FTIR will allow us to assess biochemical changes that occur before clinical signs, indicating whether changes in therapy or interventions are necessary.

Attenuated Total Reflection-Fourier Transform Infrared (ATR-FTIR) Spectroscopy Analysis of Saliva for Breast Cancer Diagnosis.

Saliva biomarkers using reagent-free biophotonic technology have not been investigated as a strategy for early detection of breast cancer (BC). The attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy has been proposed as a promising tool for disease diagnosis. However, its utilization in cancer is still incipient, and currently saliva has not been used for BC screening. We have applied ATR-FTIR onto saliva from patients with breast cancer, benign breast disease, and healthy matched controls to investigate its potential use in BC diagnosis. Several salivary vibrational modes have been identified in original and second-derivative spectra. The absorbance levels at wavenumber 1041 cm-1 were significantly higher (p < 0.05) in saliva of breast cancer patients compared with those of benign patients, and the ROC curve analysis of this peak showed a reasonable accuracy to discriminate breast cancer from benign and control patients. The 1433-1302.9 cm-1 band area was significantly higher (p < 0.05) in saliva of breast cancer patients than in control and benign patients. This salivary ATR-FTIR spectral area was prevalidated as a potential diagnostic biomarker of BC. This spectral biomarker was able to discriminate human BC from controls with sensitivity and specificity of 90% and 80%, respectively. Besides, it was able to differentiate BC from benign disease with sensitivity and specificity of 90% and 70%, respectively. Briefly, for the first time, saliva analysis by ATR-FTIR spectroscopy has demonstrated the potential use of salivary spectral biomarkers (1041 cm-1 and 1433-1302.9 cm-1) as a novel alternative for noninvasive BC diagnosis, which could be used for screening purposes.

Potential application value of Doppler ultrasonography in neoadjuvant chemotherapy for breast cancer.

BACKGROUND: The aim of this study was to evaluate the efficacy of the ultrasonography with Power Doppler, by quantifying the vascular density by Vascularity Index (VI), to predict the pathologic response to neoadjuvant chemotherapy (NAC) for breast cancer (BC). METHODS: For this prospective observational study, 20 patients were recruited with a histological diagnosis of infiltrating breast carcinoma and indication for NAC. Patients were submitted to ultrasonography with Power Doppler. Tumor vascular density was evaluated by computer software SysArea©1, before treatment, after 2 or 3 cycles of chemotherapy and at the end of treatment. The pathologic response was analyzed. The sensitivity, specificity and positive and negative predictive values for the histological response were also calculated and P-values <0.05. RESULTS: VI showed a sensitivity of 88.88%, specificity of 100%, positive predictive value of 100% and negative predictive value of 91.66% for the pathologic response, as well as being strongly associated with it. The variation of VI, in terms of either an increase or decrease after 2 or 3 cycles of chemotherapy, significantly predicted the final histological response. CONCLUSIONS: The analysis of VI was predictive and showed a strong correlation with histological response to neoadjuvant chemotherapy for BC.

A novel highly reactive Fab antibody for breast cancer tissue diagnostics and staging also discriminates a subset of good prognostic triple-negative breast cancers.

The discovery of novel markers for breast cancer (BC) has been recently relied on antibody combinatorial libraries and selection through phage display. We constructed a recombinant Fab library, and after selections against BC tissues, the FabC4 clone was thoroughly investigated by immunohistochemistry in 232 patients with long-term follow-up. The FabC4 ligand was determined by mass spectrometry. The FabC4 expression was associated with younger age, lack of progesterone receptor, higher histological grades and non-luminal subtypes, and it also identified a subset of good prognostic triple-negative BCs, possibly targeting a conformational epitope of Cytokeratin-10 (CK10). This new CK10-epitope specific antibody may open new possibilities in diagnostic and therapeutic strategies.