Intra-operative spectroscopic assessment of surgical margins during breast conserving surgery.

BACKGROUND: In over 20% of breast conserving operations, postoperative pathological assessment of the excised tissue reveals positive margins, requiring additional surgery. Current techniques for intra-operative assessment of tumor margins are insufficient in accuracy or resolution to reliably detect small tumors. There is a distinct need for a fast technique to accurately identify tumors smaller than 1 mm2 in large tissue surfaces within 30 min. METHODS: Multi-modal spectral histopathology (MSH), a multimodal imaging technique combining tissue auto-fluorescence and Raman spectroscopy was used to detect microscopic residual tumor at the surface of the excised breast tissue. New algorithms were developed to optimally utilize auto-fluorescence images to guide Raman measurements and achieve the required detection accuracy over large tissue surfaces (up to 4 × 6.5 cm2). Algorithms were trained on 91 breast tissue samples from 65 patients. RESULTS: Independent tests on 121 samples from 107 patients - including 51 fresh, whole excision specimens - detected breast carcinoma on the tissue surface with 95% sensitivity and 82% specificity. One surface of each uncut excision specimen was measured in 12-24 min. The combination of high spatial-resolution auto-fluorescence with specific diagnosis by Raman spectroscopy allows reliable detection even for invasive carcinoma or ductal carcinoma in situ smaller than 1 mm2. CONCLUSIONS: This study provides evidence that this multimodal approach could provide an objective tool for intra-operative assessment of breast conserving surgery margins, reducing the risk for unnecessary second operations.

Correlation between Ki-67 Expression and Hemodynamics of Contrast-Enhanced Ultrasound in Patients with Breast Infiltrative Ductal Carcinoma.

Breast cancer causes great threats to public health worldwide. The aim of this study was to investigate the correlation between Ki-67 expression and the hemodynamics of contrast-enhanced ultrasound (CEUS) in patients with infiltrative ductal carcinoma (IDC). CEUS was performed on 109 masses in 85 IDC cases before resection. Based on the immunohistochemical staining on the antigen Ki-67, the masses were divided into negative group, weakly positive group, positive group, and strong-positive group. Significant statistical differences were noticed in time to peak, arrive intensity, and peak intensity in the positive groups compared with the negative group. Compared with the positive groups, the negative group showed significant statistical differences in arrive intensity and peak intensity. The antigen Ki-67 was positively correlated with arrived intensity, intensity changes, and rising curve's slope. In contrast, it was negatively correlated with arrived time, time to peak, and continuous time. The hemodynamic parameters of CEUS were correlated with the expression of antigen Ki-67. On this basis, Ki-67 is an effective supplement to the diagnosis of IDC.

Correlation between elastic parameters and collagen fibre features in breast lesions.

AIM: To correlate elastic parameters with collagen fibre shape and arrangement features in breast lesions. MATERIALS AND METHODS: Shear-wave elastography (SWE) was used to measure the stiffness of breast lesions in 54 patients before surgical removal. The value of stiffness was expressed as the mean and maximum elasticity (Emean and Emax). Lesions were sliced and stained with picric acid-sirius red to display the extracellular matrix (ECM) collagen fibre. The categories of the collagen fibres were based on the shape and arrangement features, i.e., category 0, wavy collagen fibres similar to normal breast tissue; category 1, taut parallel collagen fibres around tumour nests; category 2, straightened and aligned collagen fibres tending to be perpendicular to the tumour boundary; category 3, collagen fibre in a honeycomb arrangement. Bivariate correlation analysis was used to analyse the relationship between elastic parameters and collagen fibre category. RESULTS: For all 54 lesions, the correlation coefficient between Emean and category was 0.693 (p < 0.001), and between Emax and category was 0.794 (p < 0.001). For 36 malignant lesions, the correlation coefficient between Emean and category was 0.658 (p < 0.001), and between Emax and category was 0.771 (p < 0.001). CONCLUSION: Emean and Emax of breast lesions evaluated by SWE were positively correlated with ECM collagen fibre shape and arrangement category. Changes of ECM collagen fibre shape and arrangement may account for the stiffness variations of breast lesions.

SOX11 promotes invasive growth and ductal carcinoma in situ progression.

Here, we show that SOX11, an embryonic mammary marker that is normally silent in postnatal breast cells, is expressed in many oestrogen receptor-negative preinvasive ductal carcinoma in situ (DCIS) lesions. Mature mammary epithelial cells engineered to express SOX11 showed alterations in progenitor cell populations, including an expanded basal-like population with increased aldehyde dehydrogenase (ALDH) activity, and increased mammosphere-forming capacity. DCIS.com cells engineered to express SOX11 showed increased ALDH activity, which is a feature of cancer stem cells. The CD44+/CD24-/ALDH+ cell population was increased in DCIS.com cells that expressed SOX11. Upregulating SOX11 expression in DCIS.com cells led to increased invasive growth both in vitro and when they were injected intraductally in a mouse model of DCIS that recapitulates human disease. Invasive lesions formed sooner and tumour growth was augmented in vivo, suggesting that SOX11 contributes to the progression of DCIS to invasive breast cancer. We identified potential downstream effectors of SOX11 during both microinvasive and invasive tumour growth stages, including several with established links to regulation of progenitor cell function and prenatal developmental growth. Our findings suggest that SOX11 is a potential biomarker for DCIS lesions containing cells harbouring distinct biological features that are likely to progress to invasive breast cancer. © 2017 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.

Effect of exercise training and detraining in autonomic modulation and cardiorespiratory fitness in breast cancer survivors.

BACKGROUND: Breast cancer is one of the leading causes of death worldwide. Heart rate variability (HRV) has attracted scientific community attention in different pathologies, becoming thus an ultimate importance tool in both clinical and research setting, being a good predictor of cardiac events and mortality risk and also used in physical exercise and sports in general. The aim of the present study was to evaluate 12 weeks of exercise training and six weeks of detraining in cardiorespiratory capacity, and autonomic modulation in breast cancer patients. METHODS: The sample was composed of 18 females (9 controls and 9 exercised), (aged 30-60 years). The HRV in the time and frequency domain was performed using an electrocardiogram before, after 12 weeks of the session of exercise training and after six weeks of detraining. Shapiro-Wilk and Mann-Whitney tests were made. RESULTS: No significant changes in time domain were found. In the frequency domain, 12 weeks of exercise training promote a decrease in LF (nu) and decrease in HF (nu) Index. The exercise training period promoted a decrease in LF/HF. The autonomic data returned to baseline levels after the detraining period. However, cardiorespiratory capacity remained increased after the detraining period. CONCLUSIONS: These data demonstrated that exercise training can be used to prevent autonomic dysfunction in breast cancer patients, but detraining promotes loss of all autonomic benefits.

Sentinel lymph node biopsy and aberrant lymphatic drainage in recurrent breast cancer: Findings likely to change treatment decisions.

BACKGROUND AND OBJECTIVES: It is not clear whether sentinel lymph node biopsy (SLNB) can be applied to patients with a second breast cancer or recurrence occurring at previously treated breast. The purpose of this study was to assess the feasibility of SLNB procedure in patients with recurrent breast cancer. METHODS: Patients with non-metastatic recurrent N0 breast cancer at ipsilateral breast were included. Patients were grouped according to their initial breast, axilla, and overall surgery. Presence of drainage and its pattern as well as SLNB success rate and overall axillary involvement rates were assessed. Findings were compared. RESULTS: Out of 75 patients, mean age was 52.5 years and disease-free interval was 82 (9-312) months. Lymphatic drainage was successful in 42 (56%) patients. Drainage positivity was more frequent in patients who were previously treated with SLNB (82.6%) than in patients who underwent axillary lymph node dissection (ALND) (44.2%; P = 0,002). Aberrant lymphatic drainage was detected in 64.3% of drainage positive patients. Success rate of reoperative SLNB was 92.9%. Adjuvant treatment plan was altered in 12 (16%) patients. In 15 patients, negative SLNB prevented axillary dissection. CONCLUSIONS: Reoperative SLNB seems to be technically feasible in N0 recurrent breast cancer patients. It may further avoid unnecessary ALND and lead changes in adjuvant treatment plans. J. Surg. Oncol. 2016;114:796-802. © 2016 2016 Wiley Periodicals, Inc.

Clinico-epidemiological profile of breast cancer patients and the retrospective application of Gail model 2: An Indian perspective.

AIM: To study the clinical and epidemiological profile of patients of breast cancer presenting at our center at New Delhi, India and to evaluate the applicability of Gail model 2 as a means of measuring 5-year and lifetime risk in our already diagnosed cases of breast cancer. METHODS: This was a retrospective study conducted at Lady Hardinge Medical College Hospital in New Delhi, India, between January 2011 and July 2014. Two hundred and twenty two diagnosed cases of breast cancer were included. Information was collected retrospectively on a Performa from the medical record section and the Pathology department of the hospital.The predicted five-year and lifetime risk was calculated using GM2 prediction model from the NCI's breast cancer risk assessment tool website. RESULTS AND CONCLUSIONS: Breast cancer in India is a far more biologically aggressive disease than in the west with a widely different spectrum of presentation and behavior and late presentation in an advanced stage. The accepted risk factors routinely associated with breast cancer in western literature do not appear to be relevant in the Indian population. Accepted western models do not seem to apply in the Indian scenario.

Differentiation of mammary tumors and reduction in metastasis upon Malat1 lncRNA loss.

Genome-wide analyses have identified thousands of long noncoding RNAs (lncRNAs). Malat1 (metastasis-associated lung adenocarcinoma transcript 1) is among the most abundant lncRNAs whose expression is altered in numerous cancers. Here we report that genetic loss or systemic knockdown of Malat1 using antisense oligonucleotides (ASOs) in the MMTV (mouse mammary tumor virus)-PyMT mouse mammary carcinoma model results in slower tumor growth accompanied by significant differentiation into cystic tumors and a reduction in metastasis. Furthermore, Malat1 loss results in a reduction of branching morphogenesis in MMTV-PyMT- and Her2/neu-amplified tumor organoids, increased cell adhesion, and loss of migration. At the molecular level, Malat1 knockdown results in alterations in gene expression and changes in splicing patterns of genes involved in differentiation and protumorigenic signaling pathways. Together, these data demonstrate for the first time a functional role of Malat1 in regulating critical processes in mammary cancer pathogenesis. Thus, Malat1 represents an exciting therapeutic target, and Malat1 ASOs represent a potential therapy for inhibiting breast cancer progression.

Barcoding reveals complex clonal dynamics of de novo transformed human mammary cells.

Most human breast cancers have diversified genomically and biologically by the time they become clinically evident. Early events involved in their genesis and the cellular context in which these events occur have thus been difficult to characterize. Here we present the first formal evidence of the shared and independent ability of basal cells and luminal progenitors, isolated from normal human mammary tissue and transduced with a single oncogene (KRAS(G12D)), to produce serially transplantable, polyclonal, invasive ductal carcinomas within 8 weeks of being introduced either subrenally or subcutaneously into immunodeficient mice. DNA barcoding of the initial cells revealed a dramatic change in the numbers and sizes of clones generated from them within 2 weeks, and the first appearance of many 'new' clones in tumours passaged into secondary recipients. Both primary and secondary tumours were phenotypically heterogeneous and primary tumours were categorized transcriptionally as 'normal-like'. This system challenges previous concepts that carcinogenesis in normal human epithelia is necessarily a slow process requiring the acquisition of multiple driver mutations. It also presents the first description of initial events that accompany the genesis and evolution of malignant human mammary cell populations, thereby contributing new understanding of the rapidity with which heterogeneity in their properties can develop.

Coexistence of metastatic breast carcinoma and primary tuberculosis in axillary lymph nodes: a report of a rare case.

Concomitant breast cancer metastasis and tubercular lymphadenitis in axillary lymph node is an extremely rare occurrence. Axillary lymph node metastasis is the most important factor in the staging of breast carcinoma and the number of axillary nodes showing metastases alters the stage. As tuberculosis also produces nodal enlargement, this can mimic or complicate the staging of malignant disease. Dual pathology in an organ can lead to difficulties in interpretation and inappropriate treatment of tuberculosis as well as carcinoma breast. Moreover, fine needle aspiration cytology (FNAC) from such cases may be misleading if only one of the diseases is picked up. Therefore, the need for multiple attempts at FNAC should be stressed upon for all palpable lumps. We report a case of infiltrating duct carcinoma breast in a 45-year-old female where tuberculosis was discovered in axillary lymph nodes in addition to metastases. As the present case led to incidental discovery of tuberculosis with tumor metastasis, it reinforces the possibility of a coexistent lesion in the pathologists' mind, especially in regions endemic for tuberculosis.

Estrogen regulates Hippo signaling via GPER in breast cancer.

The G protein-coupled estrogen receptor (GPER) mediates both the genomic and nongenomic effects of estrogen and has been implicated in breast cancer development. Here, we compared GPER expression in cancerous tissue and adjacent normal tissue in patients with invasive ductal carcinoma (IDC) of the breast and determined that GPER is highly upregulated in cancerous cells. Additionally, our studies revealed that GPER stimulation activates yes-associated protein 1 (YAP) and transcriptional coactivator with a PDZ-binding domain (TAZ), 2 homologous transcription coactivators and key effectors of the Hippo tumor suppressor pathway, via the Gαq-11, PLCß/PKC, and Rho/ROCK signaling pathways. TAZ was required for GPER-induced gene transcription, breast cancer cell proliferation and migration, and tumor growth. Moreover, TAZ expression positively correlated with GPER expression in human IDC specimens. Together, our results suggest that the Hippo/YAP/TAZ pathway is a key downstream signaling branch of GPER and plays a critical role in breast tumorigenesis.

Multiple myeloma or metastatic carcinoma breast: diagnostic dilemma in a case presenting with lytic bony lesion.

Carcinoma breast is common tumor which tends to metastasize to different organs but diagnostic difficulty may arise if known case of carcinoma eventually presents with another secondary malignancy with clinical features which closely mimics with metastatic carcinoma. This overlapping of symptoms may lead to delayed diagnosis of secondary malignancy with worsening of the condition of patient due to lack of specific treatment.The present case is being reported where a known case of primary breast carcinoma presented with scapular erosions suggesting metastasis. However, despite treatment her worsening condition necessitated further work up revealing multiple myeloma. The case signifies the pitfall due to overlapping of symptoms and draws attention to the fact that every known case of primary carcinoma suggestive of metastasis should also be investigated in light of another secondary malignancy. Early diagnosis of secondary malignancy followed by specific treatment would be helpful in improved prognosis of the patient.

Effect of menstrual cycle and menopausal status on apparent diffusion coefficient values and detectability of invasive ductal carcinoma on diffusion-weighted MRI.

The purpose of this study was to determine whether the apparent diffusion coefficient (ADC) and tumor detectability based on diffusion-weighted imaging (DWI) are affected by the menstrual cycle or menopausal status in breast cancer patients. Institutional review board approval was obtained, and the requirement for informed consent was waived. A total of 124 women with invasive ductal carcinoma not otherwise specified (IDC NOS) who underwent breast MRI with DWI were included in this study. Two radiologists retrospectively measured the ADCs of tumor and contralateral normal glandular tissue and scored the tumor detectability. The ADCs and detectability were compared to menstrual cycle and menopausal status, based on patient questionnaires. ADCs of tumors and contralateral tissue were significantly lower in postmenopausal women than in premenopausal women (P = 0.006 and P < 0.001, respectively). Tumor detectability did not differ significantly between the premenopausal and postmenopausal groups (P = 0.454). Normalized ADCs were not significantly lower in postmenopausal women compared to premenopausal women (P = 0.880). There was no statistically significant difference in the absolute, contralateral, and normalized ADCs (P = 0.091, 0.809, and 0.299, respectively), and the tumor detectability (P = 0.680) according to the menstrual cycle. Although ADCs of the IDC and normal glandular tissue in postmenopausal women were significantly lower than those in premenopausal women, the menstrual cycle did not affect tumor detectability and ADCs of IDC.

The Female Sexual Functioning Index (FSFI): evaluation of acceptability, reliability, and validity in women with breast cancer.

PURPOSE: Sexual dysfunction commonly arises for women following diagnosis and treatment of breast cancer. The aim of this study was to systematically evaluate the acceptability, reliability, and validity of the Female Sexual Functioning Index (FSFI) when used with these women. METHODS: Sexually active women previously diagnosed with breast cancer (N = 399) completed an online questionnaire including the FSFI and measures of acceptability (ease of use, relevance), sexual functioning, body image, fatigue, impact of cancer, physical and mental health, and relationship adjustment. Reliability and validity were evaluated using standard scale validation techniques. RESULTS: Participants indicated a high degree of acceptability. Excellent internal consistency (α = 0.83-0.96) and test-retest reliability (r = 0.74-0.86) of the FSFI were evident. According to the confirmatory factor analysis, the best fit was achieved with removal of item 14 (regarding the extent of emotional closeness with the partner) and six subscales (desire, arousal, lubrication, orgasm, satisfaction, pain), without a total score (TLI = 0.96, CFI = 0.97, RMSEA = 0.07). Correlations with measures of sexual functioning and related constructs provided evidence for convergent and divergent validities, respectively. All but one subscale (orgasm) discriminated between women who are, and are not, currently receiving treatment for breast cancer (discriminant validity). CONCLUSIONS: These findings indicate that not only is the FSFI psychometrically sound when used with women with breast cancer, but it is perceived as being easy to use and relevant. It is recommended that the FSFI subscale scores can be used in both clinical and research settings as a screening tool to identify women experiencing sexual dysfunction following breast cancer.

P2X7 receptor-mediated analgesia in cancer-induced bone pain.

Pain is a common and debilitating complication for cancer patients significantly compromising their quality of life. Cancer-induced bone pain involves a complex interplay of molecular events, including mechanisms observed in inflammatory and neuropathic pain states, but also changes unique for cancer-induced bone pain. The P2X7 receptor (P2X7R) is involved in a variety of cellular functions and has been linked to both inflammatory and neuropathic pain. Here we study the analgesic potential of P2X7R antagonism in a rat model of cancer-induced bone pain. In cancer-bearing animals, the P2X7R antagonist A839977 attenuated dorsal horn neuronal responses in a modality and intensity-specific way. Spinal application of 0.4-mg/kg and 1.2-mg/kg A839977 significantly reduced the evoked responses to high-intensity mechanical and thermal stimulation, whereas no effect was seen in response to low-intensity or electrical stimulation. In contrast, A839977 had no effect on the tested parameters in naïve or sham animals. In awake animals, 40-mg/kg A839977 (i.p.) significantly reduced both early- and late-stage pain behavior. In contrast, no effect was observed in sham or vehicle-treated animals. The results suggest that the P2X7R is involved in the mechanisms of cancer-induced bone pain, and that P2X7R antagonism might be a useful analgesic target. No effect was observed in sham or naïve animals, indicating that the P2X7R-mediated effect is state-dependent, and might therefore be an advantageous target compared to traditional analgesics.

MCP-1 stimulates spinal microglia via PI3K/Akt pathway in bone cancer pain.

Accumulating evidence suggests that chemokine monocyte chemoattractant protein-1 (MCP-1) is significantly involved in the activation of spinal microglia associated with pathological pain, at the same time that the phosphatidylinositol 3-kinase/Protein Kinase B (PI3K/Akt) pathway localized in spinal microglia is involved in both neuropathic and inflammatory pain. However, whether there is a connection between MCP-1 and the PI3K/Akt pathway and in their underlying mechanisms in bone cancer pain (BCP) has not yet been elucidated. In the current study, we investigated the expression changes of p-Akt in microglia and OX-42 (microglia marker) after being stimulated with MCP-1 in vitro, as well as in a BCP model that was established by an intramedullary injection of mammary gland carcinoma cells(Walker 256 cells) into the tibia of rats. We observed a significant increase in expression levels of p-Akt and OX-42 in microglia as well as in spinal dorsal horns of BCP rats. Furthermore, the intrathecal administration of an anti-MCP-1 neutralizing antibody or PI3K inhibitor LY294002 reduced the expression of p-Akt or OX-42, and LY294002 attenuated the mechanical allodynia of BCP rats. These results suggest that MCP-1 may stimulate spinal microglia via the PI3K/Akt pathway in BCP.

The potential role of estrogen receptor ß2 in breast cancer.

Endocrine therapy is provided to all patients with estrogen receptor (ER)-positive breast cancer, but only a subset of them derives clinical benefit. The discovery of ERß and its five isoforms added another layer of complexity in the regulation of estrogen activity in breast cancer cells. Two large retrospective studies showed conflicting results with regard to the prognostic value of the different ERß isoforms in patients treated with tamoxifen in an adjuvant setting. This study tested the hypothesis that ERß1 and, or ERß2 are correlated with clinical outcome. We identified patients with breast cancer who had undergone surgery at Bucheon St. Mary's Hospital, the Catholic University of Korea, between January 2004 and March 2006. We evaluated 101 consecutive cases for ERß1 and ERß2 expression using immunohistochemical staining and obtained other clinicopathology by reviewing medical records. ERß1 was expressed in 81.2% (79 of 97) and ERß2 was expressed in 50.5% (51 of 101) of primary breast cancer tissues. Disease-free survival (DFS) and overall survival (OS) of patients with cancers expressing ERß2 was significantly worse. Moreover, in subgroup analysis according to the tamoxifen treatment, ERß2 expression was significantly associated with shorter DFS of tamoxifen-treated patients. This study indicates that breast cancer with ERß2 expression was associated with worse DFS and OS, especially in tamoxifen treated patients. Our results suggest a role for ERß2 as an independent prognostic marker and might serve as a new therapeutic target.

Lobular neoplasia: morphology and management.

CONTEXT: Lobular neoplasia encompasses a spectrum of disease, including atypical lobular hyperplasia and lobular carcinoma in situ. Although classic forms of lobular neoplasia are predominantly heralded as a risk marker, the pleomorphic form of lobular carcinoma in situ is generally regarded as a more aggressive subtype and a possible cancer precursor, and thus is treated in a manner more similar to ductal carcinoma in situ than classic forms of lobular neoplasia. OBJECTIVE: To focus on the morphologic spectrum of lobular neoplasia as highlighted by 3 cases and current management recommendations. Areas of diagnostic challenge and controversy are addressed. DATA SOURCES: A review of the pertinent published literature and current national guidelines was conducted. CONCLUSIONS: Correct classification of classic lobular neoplasia and pleomorphic lobular carcinoma in situ is critical because of differences in clinical management, with current treatment strategies focused on risk reduction for patients with classic lobular neoplasia and eradication of the lesion for those with pleomorphic lobular carcinoma in situ.

Lapatinib and platinum-based chemotherapy ameliorate breast cancer with choroidal metastasis and restore visual acuity.

Involvement of the central nervous system in metastatic breast cancer is a particularly dismal occurrence because of its effects on mortality and quality of life. Development of choroidal metastasis in a breast cancer patient indicates poor prognosis and has become the major life-limiting problem. Various treatment modalities for choroidal metastasis have been applied with different efficacy. Here we describe a patient with HER2-overexpressing breast cancer and limited choroidal metastasis who responded to an HER2 tyrosine kinase inhibitor after failure of radiotherapy and chemotherapy. Her visual acuity was completely and durably restored after the targeted therapy. This case provides a unique treatment experience of breast cancer with choroidal metastasis.