Afadin cooperates with Claudin-2 to promote breast cancer metastasis.

Claudin-2 promotes breast cancer liver metastasis by enabling seeding and early cancer cell survival. We now demonstrate that the PDZ-binding motif of Claudin-2 is necessary for anchorage-independent growth of cancer cells and is required for liver metastasis. Several PDZ domain-containing proteins were identified that interact with the PDZ-binding motif of Claudin-2 in liver metastatic breast cancer cells, including Afadin, Arhgap21, Pdlim2, Pdlim7, Rims2, Scrib, and ZO-1. We specifically examined the role of Afadin as a potential Claudin-2-interacting partner that promotes breast cancer liver metastasis. Afadin associates with Claudin-2, an interaction that requires the PDZ-binding motif of Claudin-2. Loss of Afadin also impairs the ability of breast cancer cells to form colonies in soft agar and metastasize to the lungs or liver. Immunohistochemical analysis of Claudin-2 and/or Afadin expression in 206 metastatic breast cancer tumors revealed that high levels of both Claudin-2 and Afadin in primary tumors were associated with poor disease-specific survival, relapse-free survival, lung-specific relapse, and liver-specific relapse. Our findings indicate that signaling downstream from a Claudin-2/Afadin complex enables the efficient formation of breast cancer metastases. Moreover, combining Claudin-2 and Afadin as prognostic markers better predicts the potential of breast cancer to metastasize to soft tissues.

Progressive polarity loss and luminal collapse disrupt tissue organization in carcinoma.

Epithelial cancers (carcinoma) account for 80%-90% of all cancers. The development of carcinoma is associated with disrupted epithelial organization and solid ductal structures. The mechanisms underlying the morphological development of carcinoma are poorly understood, but it is thought that loss of cell polarity is an early event. Here we report the characterization of the development of human breast lesions leading to carcinoma. We identified a unique mechanism that generates solid ducts in carcinoma through progressive loss of polarity and collapse of the luminal architecture. This program initiates with asymmetric divisions of polarized cells that generate a stratified epithelium containing both polarized and depolarized cells. Stratified regions form cords that penetrate into the lumen, subdividing it into polarized secondary lumina. The secondary lumina then collapse with a concomitant decrease in RhoA and myosin II activity at the apical membrane and ultimately lose apical-basal polarity. By restoring RhoA activity in mice, ducts maintained lumen and cell polarity. Notably, disrupted tissue architecture through luminal collapse was reversible, and ducts with a lumen were re-established after oncogene suppression in vivo. This reveals a novel and common mechanism that contributes to carcinoma development by progressively disrupting cell and tissue organization.

Infiltration of CD8+ T cells into tumor cell clusters in triple-negative breast cancer.

Infiltration of [Formula: see text] T lymphocytes into solid tumors is associated with good prognosis in various types of cancer, including triple-negative breast cancer (TNBC). However, the mechanisms underlying different infiltration levels are largely unknown. Here, we have characterized the spatial profile of [Formula: see text] T cells around tumor cell clusters (tightly connected tumor cells) in the core and margin regions in TNBC patient samples. We found that in some patients, the [Formula: see text] T cell density first decreases when moving in from the boundary of the tumor cell clusters and then rises again when approaching the center. To explain various infiltration profiles, we modeled the dynamics of T cell density via partial differential equations. We spatially modulated the diffusion/chemotactic coefficients of T cells (to mimic physical barriers) or introduced the localized secretion of a diffusing T cell chemorepellent. Combining the spatial-profile analysis and the modeling led to support for the second idea; i.e., there exists a possible chemorepellent inside tumor cell clusters, which prevents [Formula: see text] T cells from infiltrating into tumor cell clusters. This conclusion was consistent with an investigation into the properties of collagen fibers which suggested that variations in desmoplastic elements does not limit infiltration of [Formula: see text] T lymphocytes, as we did not observe significant correlations between the level of T cell infiltration and fiber properties. Our work provides evidence that [Formula: see text] T cells can cross typical fibrotic barriers and thus their infiltration into tumor clusters is governed by other mechanisms possibly involving a local repellent.

Value of Shear Wave Elastography for the Differentiation of Benign and Malignant Microcalcifications of the Breast.

OBJECTIVE. The objective of our study was to evaluate whether shear wave elastography (SWE) can differentiate benign from malignant microcalcifications of the breast when detected on ultrasound (US). SUBJECTS AND METHODS. Between February 9, and June 23, 2016, 74 patients with mammographically detected suspicious microcalcifications underwent breast US. When microcalcifications were identified on US, stiffness was assessed using SWE. Biopsy was subsequently performed under US guidance using a 10-gauge vacuum-assisted needle. Qualitative and quantitative elastography results were compared between benign and malignant calcifications as well as between pure ductal carcinoma in situ and lesions with invasive components using the Mann-Whitney U test. ROC curves were created to assess the performance of SWE in detecting malignancy and invasive components. RESULTS. Twenty-nine groups of microcalcifications in 29 patients were identified on US. At pathology, 16 groups were benign and 13 were malignant. Stiffness of malignant calcifications was significantly higher than that of the benign ones (p = 0.0004). The AUC, sensitivity, specificity, negative predictive value, positive predictive value, and accuracy of SWE for the diagnosis of malignancy were 0.89, 69%, 100%, 80%, 100%, and 86%, respectively, and for detection of an invasive component were 0.93, 75%, 100%, 75%, 100%, and 85%. CONCLUSION. SWE has the potential to differentiate benign from malignant micro-calcifications of the breast when detected on US with high specificity.

Features from Computerized Texture Analysis of Breast Cancers at Pretreatment MR Imaging Are Associated with Response to Neoadjuvant Chemotherapy.

Purpose To evaluate whether features from texture analysis of breast cancers were associated with pathologic complete response (pCR) after neoadjuvant chemotherapy and to explore the association between texture features and tumor subtypes at pretreatment magnetic resonance (MR) imaging. Materials and Methods Institutional review board approval was obtained. This retrospective study included 85 patients with 85 breast cancers who underwent breast MR imaging before neoadjuvant chemotherapy between April 10, 2008, and March 12, 2015. Two-dimensional texture analysis was performed by using software at T2-weighted MR imaging and contrast material-enhanced T1-weighted MR imaging. Quantitative parameters were compared between patients with pCR and those with non-pCR and between patients with triple-negative breast cancer and those with non-triple-negative cancer. Multiple logistic regression analysis was used to determine independent parameters. Results Eighteen tumors (22%) were triple-negative breast cancers. pCR was achieved in 30 of the 85 tumors (35%). At univariate analysis, mean pixel intensity with spatial scaling factor (SSF) of 2 and 4 on T2-weighted images and kurtosis on contrast-enhanced T1-weighted images showed a significant difference between triple-negative breast cancer and non-triple-negative breast cancer (P = .009, .003, and .001, respectively). Kurtosis (SSF, 2) on T2-weighted images showed a significant difference between pCR and non-pCR (P = .015). At multiple logistic regression, kurtosis on T2-weighted images was independently associated with pCR in non-triple-negative breast cancer (P = .033). A multivariate model incorporating T2-weighted and contrast-enhanced T1-weighted kurtosis showed good performance for the identification of triple-negative breast cancer (area under the receiver operating characteristic curve, 0.834). Conclusion At pretreatment MR imaging, kurtosis appears to be associated with pCR to neoadjuvant chemotherapy in non-triple-negative breast cancer and may be a promising biomarker for the identification of triple-negative breast cancer. © RSNA, 2017.

Immunohistochemistry Microarrays.

Immunohistochemistry (IHC) on tissue sections is widely used for quantifying the expression patterns of proteins and is part of the standard of care for cancer diagnosis and prognosis, but is limited to staining a single protein per tissue. Tissue microarray and microfluidics staining methods have emerged as powerful high throughput techniques, but they either only permit the analysis of a single protein per slide or require complex instrumentation and expertise while only staining isolated areas. Here, we introduce IHC microarrays (IHCµA) for multiplexed staining of intact tissues with preserved histological and spatial information. Droplets of a dextran solution containing antibodies were prespotted on a slide and snapped onto a preprocessed formalin-fixed, paraffin-embedded (FFPE) tissue section soaked in a polyethylene glycol solution. The antibodies are confined within the dextran droplets and locally stain the tissue below with a contrast similar to the one obtained by conventional IHC. The microarray of antibody droplets can be prespotted on a slide and stored, thus neither the preparation of the antibody solutions nor a sophisticated microarray spotter is needed. Sampling considerations with IHCµA were evaluated by taking three tissues with varying levels of cancer cells. A multiplex IHCµA with 180 spots targeting 8 cancer proteins was performed on a breast cancer tissue section to illustrate the potential of this method. This work opens the avenue of applying microarray technologies for conducting IHC on intact tissue slices and has great potential to be used in the discovery and validation of tissue biomarkers in human tumors.

Sonographic Appearance of Lesions Diagnosed as Lobular Neoplasia at Sonographically Guided Biopsies.

OBJECTIVE: The objective of our study was to review the sonographic features of breast lesions yielding lobular neoplasia (LN) at sonographically guided biopsy, evaluate the surgical pathology outcome of these lesions, and determine if imaging findings or clinical features can be used to predict an upgrade to malignancy. MATERIALS AND METHODS: Of the 8205 sonographically guided breast biopsies (14-gauge cores) performed from 2007 through 2014, 22 yielded a diagnosis of LN, which means that LN was the most severe pathologic lesion. Imaging features were analyzed in consensus by two radiologists. Correlation of biopsy findings with definitive pathologic results was performed when available. RESULTS: Twenty-two LN lesions (20 patients [mean age ± SD, 52.05 ± 13.66 years]) were diagnosed at biopsy. Of the LN lesions that were seen on mammography (6/22, 27.3%), most lesions appeared as masses (3/6, 50%). On sonography, LN lesions (mean size, 8.10 mm) appeared as masses (15/22, 68.2%) with oval shape (10/15, 66.7%), well-circumscribed or microlobulated margins (11/15, 73.3%), hypoechoic echotexture (10/15, 66.7%), posterior enhancement (73.3%, 11/15), and parallel orientation (8/15, 53.3%). Most of the masses were categorized as BI-RADS category 4 (21/22, 95.5%). Seven lesions (7/22, 31.8%) appeared as areas of shadowing or distortion without discrete masses. Twenty (20/22, 90.9%) lesions were excised surgically, and pathology results led to an upgrade in five lesions (5/20, 25% [one nonmass lesion and four masses]). Neither mammographic nor sonographic features were associated with malignant outcome (p > 0.05). CONCLUSION: LN diagnosed at sonographically guided 14-gauge core needle biopsy does not show any specific features according to the BI-RADS lexicon and is associated with a 25% underestimation rate. No clinical or imaging characteristic is predictive of malignancy.

Differentiation of Fibroadenomas and Pure Mucinous Carcinomas on Dynamic Contrast-Enhanced MRI of the Breast Using Volume Segmentation for Kinetic Analysis: A Feasibility Study.

OBJECTIVE: The objective of this study was to retrospectively evaluate the diagnostic performance of volume-based kinetic analysis in dynamic contrast-enhanced MRI (DCE-MRI) of the breast for the differentiation of fibroadenomas (FAs) with high T2 signal intensity from pure mucinous carcinomas (PMCs). MATERIALS AND METHODS: A review of records from 2007 to 2013 that were stored in the pathology department database at our institution identified nine patients with PMCs (defined as tumor cells with a mucinous component ≥ 90%) who underwent preoperative breast MRI. The PMCs were compared with 15 biopsy-proven FAs from 13 patients. Characteristics noted on DCE-MRI were evaluated using computer-assisted diagnosis software. For each mass, the proportion of progressive enhancement in the lesion at the delayed phase was quantified. Both groups of masses were compared using a Wilcoxon signed rank test. A ROC curve was used to define an appropriate cutoff point. RESULTS: The median rate of progressive enhancement was 100% (range, 99-100%) for FAs and 97% (range, 87-99%) for PMCs (p = 0.0326). The AUC of the kinetic curve for progressive enhancement was 0.7519 (95% CI, 0.5258-0.9407). A more appropriate cutoff value to maximize sensitivity and specificity was 98.5%. With this cutoff, sensitivity was 66.7% (95% CI, 11.1-100%) and specificity was 80% (95% CI, 39.6-99.8%) for the diagnosis of PMCs. CONCLUSION: Volume-based kinetic analysis may aid in differentiating FAs from PMCs on DCE-MRI studies of the breast.

Imaging Features Associated With Posttraumatic Breast Neuromas.

OBJECTIVE: The aim of this study is to review the clinical and imaging features of patients with a histopathologic diagnosis of posttraumatic breast neuromas. MATERIALS AND METHODS: We report eight biopsy-proven posttraumatic neuromas in six patients with a history of breast surgery. Mammographic, sonographic, and breast MRI examinations were reviewed according to the BI-RADS lexicon. In addition, the tail sign, a specific sign known to be associated with neuromas in other parts of the body, was assessed. RESULTS: Mammographic, MRI, and sonographic examinations were performed in three, four and all six patients, respectively. The neuromas were occult on the three mammograms performed but all of them were identified on ultrasound as hypoechoic masses with parallel orientation; seven of the eight neuromas (87.5%) had an oval shape with circumscribed margins. Strain elastography performed for two patients (three neuromas) showed benign features (benign elasticity scores and fat-to-lesion ratio). Half of the masses showed a tail sign (focal thickening of the nerve adjacent to posttraumatic neuroma, similar to the dural tail sign). Of five lesions investigated by MRI, two were occult on MRI. The remaining three were visible as isointense foci on T1-weighted images, with a benign type 1 enhancement curve. CONCLUSION: In patients who underwent breast surgery, a mass with benign features raises the possibility of a neuroma. Although the tail sign was present in half of the posttraumatic neuromas, imaging-guided biopsy remains the standard of care.

Atypical ductal hyperplasia diagnosed at sonographically guided core needle biopsy: frequency, final surgical outcome, and factors associated with underestimation.

OBJECTIVE: The purposes of this article were to review the mammographic and sonographic features of breast masses yielding atypical ductal hyperplasia (ADH) at sonographically guided biopsy, evaluate the surgical pathology outcome of these lesions, and determine whether clinical or imaging features can be used to predict upgrade to malignancy. MATERIALS AND METHODS: Among 6325 sonographically guided biopsies (2003- 2010) (14-gauge cores), 56 yielded the diagnosis of ADH (0.9%). Six patients were excluded (lost to follow-up). Fifty lesions were surgically excised in 45 patients. Mammographic and sonographic features were analyzed in consensus by two radiologists using the BI-RADS lexicon. RESULTS: Forty-five patients (mean age, 56 years; 12 < 50 years; six with synchronous breast carcinoma) had 50 ADH lesions (median size, 0.6 cm). Surgical excision yielded malignancy in 28 cases (56% underestimation rate). Among 42 mammograms (47 lesions), 30 lesions were identified (30/47, 64%) as masses (12/30, 40%), asymmetric densities (10/30, 33%), microcalcifications (4/30, 13%), and architectural distortions (4/30, 13%). Sonographically, most lesions appeared as hypoechoic masses (64%, 30/47) with irregular shape (51%, 24/47), microlobulated margins (49%, 23/47), no posterior acoustic feature (25/47, 53%), abrupt interface (70%, 33/47), and parallel orientation (57%, 27/47). No mammographic and sonographic features were associated with malignant outcome, whereas age less than 50 years (p = 0.03) and synchronous malignancy (p = 0.03) were associated with malignant outcome. CONCLUSION: ADH diagnosed at sonographically guided 14-gauge core needle biopsy shows a high underestimation rate. Synchronous carcinoma or age less than 50 years is associated with malignant outcome.

Infiltrating syringomatous adenoma of the nipple: Sonographic and mammographic features with pathologic correlation.

We report the case of a 39-year-old woman complaining of painless unilateral nipple enlargement for 3 weeks. She had no family history of breast cancer. Clinical examination revealed left nipple enlargement without pain, erythema, or skin changes m no associated palpable breast or axillary masses. Ultrasound showed several bright foci in the left nipple suggestive of microcalcifications. Neither solid nor cystic masses were detected. The mammogram performed subsequently confirmed the presence of multiple pleomorphic microcalcifications within the nipple. Wedge biopsy showed a syringomatous adenoma. Wide local excision of the nipple was performed. The postoperative course was uneventful.

Evolution of chromosome-arm aberrations in breast cancer through genetic network rewiring.

The basal breast cancer subtype is enriched for triple-negative breast cancer (TNBC) and displays consistent large chromosomal deletions. Here, we characterize evolution and maintenance of chromosome 4p (chr4p) loss in basal breast cancer. Analysis of The Cancer Genome Atlas data shows recurrent deletion of chr4p in basal breast cancer. Phylogenetic analysis of a panel of 23 primary tumor/patient-derived xenograft basal breast cancers reveals early evolution of chr4p deletion. Mechanistically we show that chr4p loss is associated with enhanced proliferation. Gene function studies identify an unknown gene, C4orf19, within chr4p, which suppresses proliferation when overexpressed-a member of the PDCD10-GCKIII kinase module we name PGCKA1. Genome-wide pooled overexpression screens using a barcoded library of human open reading frames identify chromosomal regions, including chr4p, that suppress proliferation when overexpressed in a context-dependent manner, implicating network interactions. Together, these results shed light on the early emergence of complex aneuploid karyotypes involving chr4p and adaptive landscapes shaping breast cancer genomes.

In situ Raman spectroscopy and machine learning unveil biomolecular alterations in invasive breast cancer.

Significance: As many as 60% of patients with early stage breast cancer undergo breast-conserving surgery. Of those, 20% to 35% need a second surgery because of incomplete resection of the lesions. A technology allowing in situ detection of cancer could reduce re-excision procedure rates and improve patient survival. Aim: Raman spectroscopy was used to measure the spectral fingerprint of normal breast and cancer tissue ex-vivo. The aim was to build a machine learning model and to identify the biomolecular bands that allow one to detect invasive breast cancer. Approach: The system was used to interrogate specimens from 20 patients undergoing lumpectomy, mastectomy, or breast reduction surgery. This resulted in 238 ex-vivo measurements spatially registered with standard histology classifying tissue as cancer, normal, or fat. A technique based on support vector machines led to the development of predictive models, and their performance was quantified using a receiver-operating-characteristic analysis. Results: Raman spectroscopy combined with machine learning detected normal breast from ductal or lobular invasive cancer with a sensitivity of 93% and a specificity of 95%. This was achieved using a model based on only two spectral bands, including the peaks associated with C-C stretching of proteins around 940 cm - 1 and the symmetric ring breathing at 1004 cm - 1 associated with phenylalanine. Conclusions: Detection of cancer on the margins of surgically resected breast specimen is feasible with Raman spectroscopy.

Improved platform for breast cancer circulating tumor cell enrichment and characterization with next-generation sequencing technology.

Circulating tumor cells (CTCs) represent cells shed from the primary tumor or metastatic sites and can be used to monitor treatment response and tumor recurrence. However, CTCs circulate in extremely low numbers making in-depth analysis beyond simple enumeration challenging when collected from peripheral blood. Furthermore, tumor heterogeneity, a hallmark of many tumors, especially breast cancer, further complicates CTC characterization. To overcome this limitation, we developed a platform based on the large-scale isolation of CTCs by apheresis, allowing us to collect CTCs in large numbers, which were preserved live in liquid nitrogen for further characterization. Flow cytometry followed by cell sorting (FACS) was performed using a combination of antibodies directed against cell surface markers of white blood cells (CD45) and epithelial tumor cells (CK8). Analysis of subpopulations CD45+/- and CK8+/- by bulk RNA sequencing (RNAseq) and the CD45-/CK8 positive population by single-cell RNAseq was performed. The CD45- population was enriched using CD45 magnetic beads separation and examined by IHC for pan-cytokeratin and immunofluorescence (IF) for specific markers, including the elusive circulating cancer stem cells (CSCs). CSC-rich mammospheres were grown in vitro for further analysis and treated to examine their response to chemotherapeutic agents. Finally, mammospheres were transplanted into the mammary fat pad and bone of immunodeficient mice to examine tumor growth in vivo. This platform enables the detection and collection of CTCs in early and late-stage breast cancer patients of every subtype. Markers including CD44/24, ALDH1 and CXCR4 were identified by IF and showed high expression following mammosphere culture, which responded predictably to chemotherapeutic agents. Mammospheres were also transplanted into nude mice and induced tumors in the mammary fat pad and bone following intra-tibial transplantation. Finally, bulk RNA analysis of the FACS isolated CD45+/- and CK8+/- cells showed a clear separation of CD45- away from CD45+ populations. Single-cell RNAseq of the FACS isolated CD45-/CK8+ cells showed the presence of 4-5 clusters, confirming the high degree of heterogeneity of CTCs. Our platform for large-scale isolation of CTCs using apheresis is suitable for an in-depth analysis of the cancer phenotype and may eventually allow evaluation in real-time of the disease process to optimize cancer regimens.

ABCC5 supports osteoclast formation and promotes breast cancer metastasis to bone.

INTRODUCTION: Bone is the most common site of breast cancer metastasis, and complications associated with bone metastases can lead to a significantly decreased patient quality of life. Thus, it is essential to gain a better understanding of the molecular mechanisms that underlie the emergence and growth of breast cancer skeletal metastases. METHODS: To search for novel molecular mediators that influence breast cancer bone metastasis, we generated gene-expression profiles from laser-capture microdissected trephine biopsies of both breast cancer bone metastases and independent primary breast tumors that metastasized to bone. Bioinformatics analysis identified genes that are differentially expressed in breast cancer bone metastases compared with primary, bone-metastatic breast tumors. RESULTS: ABCC5, an ATP-dependent transporter, was found to be overexpressed in breast cancer osseous metastases relative to primary breast tumors. In addition, ABCC5 was significantly upregulated in human and mouse breast cancer cell lines with high bone-metastatic potential. Stable knockdown of ABCC5 substantially reduced bone metastatic burden and osteolytic bone destruction in mice. The decrease in osteolysis was further associated with diminished osteoclast numbers in vivo. Finally, conditioned media from breast cancer cells with reduced ABCC5 expression failed to induce in vitro osteoclastogenesis to the same extent as conditioned media from breast cancer cells expressing ABCC5. CONCLUSIONS: Our data suggest that ABCC5 functions as a mediator of breast cancer skeletal metastasis. ABCC5 expression in breast cancer cells is important for efficient osteoclast-mediated bone resorption. Hence, ABCC5 may be a potential therapeutic target for breast cancer bone metastasis.

Met induces mammary tumors with diverse histologies and is associated with poor outcome and human basal breast cancer.

Elevated MET receptor tyrosine kinase correlates with poor outcome in breast cancer, yet the reasons for this are poorly understood. We thus generated a transgenic mouse model targeting expression of an oncogenic Met receptor (Met(mt)) to the mammary epithelium. We show that Met(mt) induces mammary tumors with multiple phenotypes. These reflect tumor subtypes with gene expression and immunostaining profiles sharing similarities to human basal and luminal breast cancers. Within the basal subtype, Met(mt) induces tumors with signatures of WNT and epithelial to mesenchymal transition (EMT). Among human breast cancers, MET is primarily elevated in basal and ERBB2-positive subtypes with poor prognosis, and we show that MET, together with EMT marker, SNAIL, are highly predictive of poor prognosis in lymph node-negative patients. By generating a unique mouse model in which the Met receptor tyrosine kinase is expressed in the mammary epithelium, along with the examination of MET expression in human breast cancer, we have established a specific link between MET and basal breast cancer. This work identifies basal breast cancers and, additionally, poor-outcome breast cancers, as those that may benefit from anti-MET receptor therapies.

Intra-Tumoral CD8+ T-Cell Infiltration and PD-L1 Positivity in Homologous Recombination Deficient Pancreatic Ductal Adenocarcinoma.

The immune contexture of pancreatic ductal adenocarcinoma (PDAC) is generally immunosuppressive. A role for immune checkpoint inhibitors (ICIs) in PDAC has only been demonstrated for the rare and hypermutated mismatch repair (MMR) deficient (MMR-d) subtype. Homologous recombination repair (HR) deficient (HR-d) PDAC is more prevalent and may encompass up to 20% of PDAC. Its genomic instability may promote a T-cell mediated anti-tumor response with therapeutic sensitivity to ICIs. To investigate the immunogenicity of HR-d PDAC, we used multiplex immunohistochemistry (IHC) to compare the density and spatial distribution of CD8+ cytotoxic T-cells, FOXP3+ regulatory T-cells (Tregs), and CD68+ tumor-associated macrophages (TAMs) in HR-d versus HR/MMR-intact PDAC. We also evaluated the IHC positivity of programmed death-ligand 1 (PD-L1) across the subgroups. 192 tumors were evaluated and classified as HR/MMR-intact (n=166), HR-d (n=25) or MMR-d (n=1) based on germline testing and tumor molecular hallmarks. Intra-tumoral CD8+ T-cell infiltration was higher in HR-d versus HR/MMR-intact PDAC (p<0.0001), while CD8+ T-cell densities in the peri-tumoral and stromal regions were similar in both groups. HR-d PDAC also displayed increased intra-tumoral FOXP3+ Tregs (p=0.049) and had a higher CD8+:FOXP3+ ratio (p=0.023). CD68+ TAM expression was similar in HR-d and HR/MMR-intact PDAC. Finally, 6 of the 25 HR-d cases showed a PD-L1 Combined Positive Score of >=1, whereas none of the HR/MMR-intact cases met this threshold (p<0.00001). These results provide immunohistochemical evidence for intra-tumoral CD8+ T-cell enrichment and PD-L1 positivity in HR-d PDAC, suggesting that HR-d PDAC may be amenable to ICI treatment strategies.

Claudin-2 promotes colorectal cancer liver metastasis and is a biomarker of the replacement type growth pattern.

Claudin-2 promotes breast cancer liver metastasis by enabling seeding and early cancer cell survival. We now demonstrate that Claudin-2 is functionally required for colorectal cancer liver metastasis and that Claudin-2 expression in primary colorectal cancers is associated with poor overall and liver metastasis-free survival. We have examined the role of Claudin-2, and other claudin family members, as potential prognostic biomarkers of the desmoplastic and replacement histopathological growth pattern associated with colorectal cancer liver metastases. Immunohistochemical analysis revealed higher Claudin-2 levels in replacement type metastases when compared to those with desmoplastic features. In contrast, Claudin-8 was highly expressed in desmoplastic colorectal cancer liver metastases. Similar observations were made following immunohistochemical staining of patient-derived xenografts (PDXs) that we have established, which faithfully retain the histopathology of desmoplastic or replacement type colorectal cancer liver metastases. We provide evidence that Claudin-2 status in patient-derived extracellular vesicles may serve as a relevant prognostic biomarker to predict whether colorectal cancer patients have developed replacement type liver metastases. Such a biomarker will be a valuable tool in designing optimal treatment strategies to better manage patients with colorectal cancer liver metastases.

Ovarian Mixed Malignant Brenner-Mucinous Tumor with Signet Ring Cells.

Mucinous carcinomas with signet ring cells in the ovary, particularly those composed predominantly of signet ring cells, are extremely rare, and in vast majority of cases, they represent metastasis from another site such as the stomach, appendix, pancreaticobiliary tract, bladder, and breast (Hristov et al., 2007, Kiyokawa et al., 2006, Vang et al., 2006, Young, 2006). Malignant Brenner tumor is also rare comprising less than 0.5% of ovarian carcinoma. Although mixed Brenner-Mucinous tumors are relatively common, the combination of a primary ovarian signet ring carcinoma with a malignant Brenner tumor is unique and to the best of our knowledge not previously reported in the literature.

Epithelial-Myoepithelial Carcinoma of the Breast with Rhabdoid Features.

Epithelial-myoepithelial carcinoma of the breast is a rare biphasic tumor composed of intermixed malignant epithelial and myoepithelial components. Myoepithelial cells are known to adopt varied morphologies, including spindle, chondroid, clear cell, and rhabdoid morphologies, and can represent a diagnostic challenge when isolated on biopsy. Rhabdomyosarcoma, phyllodes tumor, metaplastic carcinoma, and myoepithelial carcinoma are primary breast tumors that all have been shown to exhibit rhabdoid features, whether representing true differentiation or morphological mimic. We here report an epithelial-myoepithelial carcinoma of the breast with rhabdoid features in a 76-year-old woman. The rhabdoid-appearing myoepithelial cells are negative for myogenin, consistent with a rhabdoid-like morphology rather than a true rhabdoid differentiation, comparably to previously described myoepithelial carcinoma with rhabdoid features. To our knowledge, this is the first reported case of epithelial-myoepithelial carcinoma of the breast with rhabdoid features and thus adds another entity to the differential diagnosis of breast lesions with rhabdoid features.