Effects of different preprocessing algorithms on the prognostic value of breast tumour microscopic images.

The purpose of this study was to improve the prognostic value of tumour histopathology image analysis methodology by image preprocessing. Key image qualities were modified including contrast, sharpness and brightness. The texture information was subsequently extracted from images of haematoxylin/eosin-stained tumour tissue sections by GLCM, monofractal and multifractal algorithms without any analytical limitation to predefined structures. Images were derived from patient groups with invasive breast carcinoma (BC, 93 patients) and inflammatory breast carcinoma (IBC, 51 patients). The prognostic performance was indeed significantly enhanced by preprocessing with the average AUCs of individual texture features improving from 0.68 ± 0.05 for original to 0.78 ± 0.01 for preprocessed images in the BC group and 0.75 ± 0.01 to 0.80 ± 0.02 in the IBC group. Image preprocessing also improved the prognostic independence of texture features as indicated by multivariate analysis. Surprisingly, the tonal histogram compression by the nonnormalisation preprocessing has prognostically outperformed the tested contrast normalisation algorithms. Generally, features without prognostic value showed higher susceptibility to prognostic enhancement by preprocessing whereas IDM texture feature was exceptionally susceptible. The obtained results are suggestive of the existence of distinct texture prognostic clues in the two examined types of breast cancer. The obtained enhancement of prognostic performance is essential for the anticipated clinical use of this method as a simple and cost-effective prognosticator of cancer outcome.

Age related influence of triple receptor status on metastatic breast cancer post relapse survival.

PURPOSE: Prognostic factors in metastatic breast cancer (MBC) differ from those of primary breast cancer. The aim of this study was to identify the clinical significance of combined estrogen and progesterone receptors (ER,PR) and human epidermal growth factor receptor-2 (HER2) status on MBC post relapse survival. METHODS: The survival of 109 MBC patients was analyzed according to clinical characteristics and ER/PR status (tested by ligand binding assay) and HER2 status (tested by chromogenic in situ hybridization/CISH). RESULTS: Proper parameters for follow up of MBC patients were patient age, duration of disease free interval (DFI), dominant site of metastasis, number of metastatic sites and ER, PR status. Follow up of MBC patients showed the statistically significant difference in post relapse survival between patients with extreme phenotypes ER+PR+ and ER-PR-. Addition of HER2 status confirmed negative effect of HER2 amplification on MBC post relapse survival resulting in worse prognosis of ER-PR-HER2+ patients. The corresponding triple receptor (ER,PR,HER2) combination repeated the same pattern. In combination with patient age it was shown that difference in post relapse survival between extreme phenotypes (ER+PR+HER2- and ER-PRHER2+) was age related i.e. patients older than 50 years, with ER-PR-HER2+ phenotype, had mortality rate 100% and median survival time 14 months. CONCLUSION: There is a strong indication for use of combined triple receptor status for follow-up of MBC patients. Based on our results, the worst phenotype was neither triple positive nor triple negative, but the one that most likely reflects the biological background of these biomarkers (ERPR- HER2+). Double and triple receptor status showed repeated pattern of influence on prognosis, but the finding that ER-PR-HER2+ phenotype in an age-restricted subgroup of patients means extremely poor prognosis and a highest mortality rate deserves further consideration regarding therapy efficiency.

The prognostic role of interleukin-8 (IL-8) and matrix metalloproteinases -2 and -9 in lymph node-negative untreated breast cancer patients.

PURPOSE: To investigate the relationships, if any, between interleukin (IL) -8/matrix metalloproteinase (MMP)-2/ MMP-9 and other prognostic variables in lymph node-negative untreated breast cancer patients, and to determine the prognostic value of these potential biomarkers. METHODS: The study included 135 patients with known clinicopathological parameters. IL-8, MMP-2 and MMP-9 levels were determined by ELISA in primary tumor tissue lysates. RESULTS: There were no significant relationships between IL-8/MMP-2/MMP-9 expression and available clinicopathological parameters (patient age, menopausal status, tumor size and tumor grade). Estrogen receptor (ER)- patients had higher levels of both IL-8 and MMP-9 (p=0.006 and p=0.04, respectively) compared to ER+ patients; there was a significant negative correlation between ER and IL-8 (p=0.02). MMP-9 expression was significantly higher in patients with higher levels of IL-8 (p<0.001) and there was a significant positive correlation between IL-8 and MMP-9, as well as between progesterone receptor (PR) and MMP-2 (p<0.001 and p=0.05, respectively). PR+ patients had higher levels of MMP-2 than PR- patients (p=0.03). Among the investigated biomarkers, only IL-8 had a statistically significant prognostic value in terms of relapse free survival (RFS) (p<0.001). Patients with higher levels of IL-8 had worse prognosis. CONCLUSIONS: Expression of IL-8 and consequently expression of MMP-9 could be hormonally regulated in breast cancer. IL-8 could be a marker of more aggressive, ER- breast cancer phenotype. Different expression of MMP-2 and MMP-9 regarding differential hormonal receptor expression could indicate distinct mechanisms of their regulation. It seems that IL-8 is a strong and independent unfavorable prognostic parameter in node-negative breast cancer. Node-negative patients with higher levels of IL-8 should be treated with adjuvant, especially IL-8 targeted therapy.

Change of influence of prognostic markers on metastasis free interval during and after adjuvant tamoxifen therapy in breast cancer patients.

PURPOSE: To evaluate the influence of molecular biomarkers (estrogen receptor - ER, progesterone receptor - PR, and human epidermal growth factor receptor2 - HER2) and pathological parameters on metastasis free interval (MFI) in adjuvantly tamoxifen-treated breast cancer patients, during different follow up periods (0-2.5 years, 2.5-5 years and 5-12 years). METHODS: The study included 113 postmenopausal breast cancer patients with known pathological parameters. Steroid receptors were determined by ligand-binding assay and HER2 amplification status by chromogenic in situ hybridization (CISH). RESULTS: During the first 2.5 years of therapy patients with ER <5 fmol/mg, PR <5 fmol/mg or pT2 (≥2cm) tumors had higher probability of distant metastasis. For the period between 2.5-5 years, analysis of MFI according to pathological parameters and molecular biomarkers, separately, did not show any statistically significant difference. Patients with pT≥2 cm and HER2 amplification had much greater chance of developing distant metastasis when compared to other phenotypes (HER2-negative/pT1, HER2-negative/pT2 and HER2-positive/pT1). Patiens with ER ≥160 fmol/mg and PR ≥45 fmol/mg had good prognosis after 5 years of tamoxifen therapy. CONCLUSION: Our study indicates that there is a change of influence of the analyzed pathological parameters on MFI, depending on different follow up periods. Steroid receptor status, tumor size and HER2 status (alone or in combination) are significant parameters for the course of disease of postmenopausal ER-positive breast cancer patients, but during different periods of follow up.

Metastatic breast cancer survival according to HER2 and Topo2a gene status.

The aim of this study was to determine the relationship between amplification of HER2 (Human epidermal growth factor receptor 2) and Topo2a (topoisomerase 2a) and their influence on prognosis in metastatic breast cancer (MBC) patients. Amplification of both HER2 and Topo2a genes was determined by chromogenic in situ hybridization (CISH) in primary tumor tissue of 71 MBC patients. Starting point for follow-up was the time of diagnosis of metastatic disease. Although there was significant correlation between HER2 amplification and Topo2a alterations, Topo2a amplification was not strictly related to HER2 amplification. Follow-up of patients showed that there was no difference in MBC survival between HER2-nonamplified and HER2-amplified patients for subgroup as whole, but there was significant difference in MBC survival between patients with and without Topo2a amplification. HER2 amplification showed prognostic value in subgroups of patients, as well as Topo2a. Combination of these two genes with different status (nonamplified, amplified, coamplified) indicated that they might have additive effect. Also, it has been shown that Topo2a-amplified cases have poorer survival than Topo2a-nonamplified, when treated with CMF therapy. Topo2a amplification seems to be more promising biomarker of MBC survival, than HER2, and potential marker of resistance to CMF therapy.

Localization of recognition site between transforming growth factor-beta1 (TGF-beta1) and TGF beta receptor type II: possible implications in breast cancer.

Although overexpression of TGF-beta1 protein has been demonstrated in advanced breast cancer (BC) patients, as well as in other solid tumours, the molecular mechanism of this process remains obscure. This paper proposes that a genetic/epigenetic alteration might occur in the TGF-beta1 gene, within the region coding for the recognition site with TGFbeta receptor type II, leading to a disruption of the ligand-receptor interaction and triggering the TGF-beta1 cascade-related BC progression. To establish the operational framework for this hypothesis, in the present study, this recognition site was identified by the Informational Spectrum Method (ISM) to comprise two TGF-beta1 peptides (positions 47-66 aa and 83-112 aa) and one receptor peptide at positions 112-151 aa of the extracellular domain of the receptor (TbetaRIIM). The TbetaRIIM locus was further evaluated by ISM-derived deletion analysis of the TbetaRII sequences. To provide experimental support for the proposed model, a pilot study of plasma TGF-beta1 analysis was performed in advanced BC patients (n = 8). Two commercial ELISA assays, one with specific alphaTGF-beta1 MAb (MAb) and other with TbetaRIIM as the immobilized phase, revealed pronounced differences in the pattern of plasma TGF-beta1 elevation. In MAb-profile, the TGF-beta1 increase was detected in 7 of 8 patients, whereas analogous TbetaRIIM-profile revealed the elevation in 3 of 8 patients, taking a 50% of maximal elevation as the cut-off value. These findings are consistent with the proposed aberration of TGF-beta1 ligand within the TbetaRII recognition site. Summarizing, this model system is a good starting point for further genetic studies, particularly on genetic/epigenetic alterations of sequences involved in TGF-beta1 and TbetaRIIM interaction, with putative prognostic value for breast cancer.

Plasma TGF-beta1-related survival of postmenopausal metastatic breast cancer patients.

A pilot study was conducted to assess whether plasma levels of transforming growth factor-beta1 (TGF-beta1) might facilitate biological subgrouping of postmenopausal metastatic breast cancer patients, and, accordingly, its applicability in clinical oncology. This study included 29 postmenopausal metastatic breast cancer patients. Plasma TGF-beta1 levels were detected by enzyme-linked immunosorbent assay (ELISA). Estrogen and progesterone receptors were assayed by radioligand binding, in accordance with the recommendation of the EORTC. Concentrations of 17-beta estradiol were determined by using ELISA-microwell method (DIALAB). Overall survival was followed for 24 months for each individual patient. Stratification of the patients by ER/PR status showed that 14 patients with estrogen receptor-negative, progesterone receptor-negative carcinomas displayed a statistically significant increase in plasma TGF-beta1 levels when compared to plasma TGF-beta1 levels of 6 patients with ER-positive, PR-positive carcinomas (P=0.04). In this study, 7 out of 14 patients with negative receptors' status had no plasma TGF-beta1 values overlapping with patients having positive receptors' status. The TGF-beta1 cut-off value was defined as the highest plasma TGF-beta1 level of ER-positive, PR-positive patients: 3.28 ng/ml. This plasma TGF-beta1 cut-off value defined low-risk subgroup of 19 patients (< or = 3.28 ng/ml) and high-risk subgroup of 10 patients (> 3.28 ng/ml) (P=0.047). Plasma TGF-beta1-related survival was independent of the classical prognostic factors of metastatic breast cancer. Accordingly, a clinical significance of elevated plasma TGF-beta1 levels may be suggested.

Natural course of node-negative breast cancer: high risk-related subgroups.

This study attempted to evaluate the first-generation prognostic factors in terms of relapse-free interval in 297 node-negative breast cancer patients treated with locoregional therapy alone. Predictors of outcome were, in order of strength: tumor size (RR 2.09), tumor grade (RR 2.03) and age (RR 0.97). Age was a single independent prognostic parameter in premenopausal patients younger than 45 (RR 0.82). Tumor size was also a single independent prognostic parameter in middle-aged patients (RR 2.05). In older patients, aged 60 and over, tumor grade (RR 11.6) and type (RR 0,52) in addition to tumor size (RR 7.00) were independent prognostic parameters. Our study of disease-free survival identified high risk-related subgroups: patients younger than 35, middle-aged post-menopausal patients bearing pT2 carcinoma with steroid receptor content lower than 5 fmol/mg, and patients older than 59 bearing pT2 carcinoma with grade 11 and unfavorable types-ductal, lobular.