Validation of prognostic and predictive value of total tumoral load after primary systemic therapy in breast cancer using OSNA assay.

PURPOSE: This study aimed to validate the classification of breast cancer (BC) patients in progression risk groups based on total tumor load (TTL) value to predict lymph node (LN) affectation after neo-adjuvant systemic therapy (NAST) obtained in the NEOVATTL study. METHODS/PATIENTS: This was an observational, retrospective, international, multicenter study including patients with infiltrating BC who received NAST followed by sentinel lymph node biopsy (SLNB) analyzed with one-step nucleic acid amplification (OSNA) from nine Spanish and two Italian hospitals. Patients were classified into three groups according to the progression risk, measured as disease-free survival (DFS), based on TTL values (> 250, 250-25,000, and > 25,000 copies/µL). The previous (NEOVATTL study) Cox regression model for prognosis was validated using prognostic index (PI) and Log ratio test (LRT) analyses; the value of TTL for axillary non-SLN affectation was assessed using receiver operating characteristic (ROC) curves. RESULTS: We included 263 patients with a mean age of 51.4 (± SD 10.5) years. Patients with TTL > 25,000 copies/µL had a shorter DFS (HR 3.561 [95% CI 1.693-7.489], p = 0.0008 vs. TTL ≤ 25,000). PI and LRT analyses showed no differences between the two cohorts (p = 0.2553 and p = 0.226, respectively). ROC analysis showed concordance between TTL and non-SLN involvement (area under the curve 0.828), with 95.7% sensitivity and 92.9% specificity at a TTL cut-off of > 15,000 copies/µL. CONCLUSIONS: In BC patients who had received NAST and underwent SLNB analysis using OSNA, a TTL value of > 25,000 copies/µL was associated with a higher progression risk and > 15,000 copies/µL was predictive of non-SLN involvement.

Breast metastasis in a young pregnant woman affected by metastatic melanoma.

Metastasis to the breast from extramammary tumors are rare, and the most common cancer that metastasizes to this site is malignant melanoma (MM). Unfortunately, metastases from malignant melanoma reveal a widespread of the disease and a high likeliness of poor diagnosis. In this study, a case of left breast metastasis of MM in a young pregnant woman, with a fast progression of the mammary and systemic course of pathology and unfortunately poor prognosis is presented. Despite the role of pregnancy in MM has yet to be unraveled, our study encourages the theory that immunosuppression and hormonal changes due to pregnancy may aggravate melanoma prognosis.

Gastro-intestinal metastasis from Melanoma: Clinico-pathological clues.

INTRODUCTION: Metastasis from melanoma in the gastro-intestinal tract is a frequent event but, in the absence of an adequate clinical context and oncological anamnesis, it could be misdiagnosed by the pathologists. Moreover, amelanotic and/or poorly differentiated metastasis from melanoma in the gastro-intestinal tract can be easily underestimated. MATERIALS AND METHODS: We describe the histological features of gastro-intestinal metastasis from melanoma in a multi-centric cohort of 49 patients. In 24/49 patients, we were able to compare histological findings such as the growth pattern and the melanotic pigment also in the primary melanoma. RESULTS: The epithelioid pattern is the most common growth pattern observed in gastro-intestinal metastasis (57 %), followed by the mixed pattern (41 %) and the spindled pattern (2 %). We documented a discordant growth pattern between metastasis and primary in 9/24 cases and the absence of melanotic pigment in 8/49 cases. DISCUSSION: Our experience highlights that pathologists should take into account the possibility of gastro-intestinal metastasis from melanoma also in cases with spindled-cells/amelanotic lesions, without a previous anamnesis of melanoma asportation, and in cases of a discordant growth pattern with the primary. A correct clinical integration and an aware immunohistochemical approach are imperative to best manage the bioptic sample in order to investigate the biological profiling and therefore plan a personalizated therapy.

Colorectal adenosquamous carcinoma: Peculiar morphologic features and distinct immunoprofiles in squamous and glandular components.

Colorectal adenosquamous carcinoma (ASC) is exceedingly rare, comprising less than 0.1% of all colorectal malignancies, and is characterized by the admixture of glandular and squamous components. Due to its rarity, immunohistochemical and biological profiles have not been well investigated. The clinico-pathologic features of 29 cases of primary colorectal adenosquamous carcinomas, including four synchronous metastases, as well as the immunohistochemical expression of keratin 20, CDX2, keratin 34ßE12, keratin 5/6, p63, p40, ß-Catenin, Cyclin D1 and mismatch repair protein (MMR) expression in both squamous and glandular components are described. All ASCs showed aggressive clinico-pathologic features; all cases showed at least one aggressive pathologic characteristic (poorly differentiated, vascular invasion, infiltrative growth pattern) and 69% of cases were either stage III or IV. The squamous component was keratin 34ßE12 positive in all cases and keratin 5/6 positive in 27 cases, while only 7 cases showed p63 and/or p40 expression. ß-Catenin and Cyclin D1 showed different expression, with nuclear staining of Cyclin D1 in the squamous component of all cases (both primary and metastatic lesions) and nuclear staining of ß-Catenin predominantly in the glandular component. All but one case showed proficient MMR profile. Sixteen patients (64%) died of their disease with median survival of 10 months. ASC show aggressive clinical outcome and aggressive pathologic characteristics. A peculiar keratin 34ßE12 positive profile in the squamous component is seen differing from squamous cell carcinoma and non-intestinal ASC. The staining patterns for ß-Catenin and Cyclin D1 between components, supports a possible divergent clonal evolution of the neoplasm.

Combined analysis of miR-200 family and its significance for breast cancer.

While the molecular functions of miR-200 family have been deeply investigated, a role for these miRNAs as breast cancer biomarkers remains largely unexplored. In the attempt to clarify this, we profiled the miR-200 family members expression in a large cohort of breast cancer cases with a long follow-up (H-CSS cohort) and in TCGA-BRCA cohort. Overall, miR-200 family was found upregulated in breast tumors with respect to normal breast tissues while downregulated in more aggressive breast cancer molecular subtypes (i.e. Luminal B, HER2 and triple negative), consistently with their function as repressors of the epithelial-to-mesenchymal transition (EMT). In particular miR-141-3p was found differentially expressed in breast cancer molecular subtypes in both H-CSS and TCGA-BRCA cohorts, and the combined analysis of all miR-200 family members demonstrated a slight predictive accuracy on H-CSS cancer specific survival at 12 years (survival c-statistic: 0.646; 95%CI 0.538-0.754).

Mediastinal Nodular Lesions Synchronous to Lung Carcinoma on Frozen Section: Trap and Lesson.

Thymoma is the most frequent neoplasm arising in the anterior mediastum. It usually presents as an enlarged central mass. In the literature, multiple thymoma is described as an unusual finding; rare variants have also been described, like the signet ring-like cell variant. Evidence of co-existence of signet ring-like cells and lymphocytes in small biopsies from nodular mediastinal lesions can lead to a diagnosis of metastatic carcinoma, mostly at frozen sections. Thymoma and pulmonary carcinoma are very rarely associated neoplasms. We present a case of two mediastinal lesions discovered during pulmonary carcinoma staging. At frozen section, a diagnosis of 'epithelioid proliferation associated to lymphoid tissue' was advanced on a sample of nodular lesions and of 'carcinoma' on pulmonary biopsy. Double AB Type Thymoma with a signet ring cell-like component, synchronous to pulmonary adenocarcinoma, was the diagnosis made on formalin fixed-paraffin embedded samples. Reporting the coexistence of these two entities can help pathologists and surgeons to establish the best management of similar patients.

Evaluation of microRNA-10b prognostic significance in a prospective cohort of breast cancer patients.

BACKGROUND: MicroRNA-10b (miR-10b) has a prominent role in regulating tumor invasion and metastasis by targeting the HOXD10 transcriptional repressor and has been found up-regulated in several tumor types. METHODS: We evaluated the expression of miR-10b in paired tumor and normal specimens obtained from a prospective cohort of breast cancer patients with at least 36 months follow-up enrolled according to the REMARK guidelines (n = 150). RNA quality was measured and only samples with RNA Integrity Number (RIN) ≥7.0 were analyzed. RESULTS: The relative expression of miR-10b in tumor as compared to its normal counterpart (RER) was determined by RT-qPCR. miR-10b RERs were higher in the subgroup of patients with synchronous metastases (n = 11, Median 0.25; IQR 0.11-1.02) as compared with patients without metastases (n = 90, Median 0.09; IQR 0.04-0.29) (p = 0.028). In the subgroup of patients without synchronous metastases (n = 90), higher miR-10b RERs were associated with increased risk of disease progression and death in both univariable (HR 1.16, p = 0.021 and HR 1.20, p = 0.015 respectively for 0.10 unitary increase of miR-10b RERs levels) and multivariable (HR1.30, p < 0.001, and HR 1.31, p = 0.003 respectively for 0.10 unitary increase of miR-10b RERs levels) Cox regression models. The addition of miR-10b RERs to the Nottingham Prognostic Index (NPI) provided an improvement in discrimination power and risk reclassification abilities for the clinical outcomes at 36 months. Survival C-indices significantly increased from 0.849 to 0.889 (p = 0.009) for OS and from 0.735 to 0.767 (p = 0.050) for DFS. CONCLUSIONS: Our results provide evidences that the addition of miR-10b RERs to the prognostic factors used in clinical routine could improve the prediction abilities for both overall mortality and disease progression in breast cancer patients.

Aberrant Keap1 methylation in breast cancer and association with clinicopathological features.

Keap1 (Kelch-like ECH-associated protein 1) is an adaptor protein that mediates the ubiquitination/degradation of genes regulating cell survival and apoptosis under oxidative stress conditions. We determined methylation status of the KEAP1 promoter in 102 primary breast cancers, 14 pre-invasive lesions, 38 paired normal breast tissues and 6 normal breast from reductive mammoplasty by quantitative methylation specific PCR (QMSP). Aberrant promoter methylation was detected in 52 out of the 102 primary breast cancer cases (51%) and 10 out of 14 pre-invasive lesions (71%). No mutations of the KEAP1 gene were identified in the 20 breast cancer cases analyzed by fluorescence based direct sequencing. Methylation was more frequent in the subgroup of patients identified as ER positive-HER2 negative tumors (66.7%) as compared with triple-negative breast cancers (35%) (p = 0.05, Chi-square test). The impact of the interactions between Er, PgR, Her2 expression and KEAP1 methylation on mortality was investigated by RECPAM multivariable statistical analysis, identifying four prognostic classes at different mortality risks. Triple-negative breast cancer patients with KEAP1 methylation had higher mortality risk than patients without triple-negative breast cancer (HR = 14.73, 95%CI: 3.65-59.37). Both univariable and multivariable COX regressions analyses showed that KEAP1 methylation was associated with a better progression free survival in patients treated with epirubicin/cyclophosfamide and docetaxel as sequential chemotherapy (HR = 0.082; 95%CI: 0.007-0.934). These results indicate that aberrant promoter methylation of the KEAP1 gene is involved in breast cancerogenesis. In addition, identifying patients with KEAP1 epigenetic abnormalities may contribute to disease progression prediction in breast cancer patients.