Linda Schiffhauer

A lower Allred score for progesterone receptor is strongly associated with a higher recurrence score of 21-gene assay in breast cancer.

ABSTRACT Among the 77 infiltrating breast carcinomas, we found that progesterone receptor (PR) expression was inversely associated with recurrence score (RS, p < .0001). RS is also significantly associated with tubule formation, mitosis, and luminal B subtype. The equation of RS = 17.489 + 2.071 (tubal formation) + 2.926 (mitosis) –2.408 (PR) –1.061 (HER2) + 7.051 (luminal A) + 29.172 (luminal B) predicts RS with an R2 of 0.65. In conclusion, PR negativity, luminal B subtype, tubal formation, and mitosis are strongly correlated with a higher RS.

C35 (C17orf37) is a novel tumor biomarker abundantly expressed in breast cancer

Identification of shared tumor-specific targets is useful in developing broadly applicable therapies. In a study designed to identify genes up-regulated in breast cancer, a cDNA clone corresponding to a novel gene C35 (C17orf37) was selected by representational difference analysis of tumor and normal human mammary cell lines. Abundant expression of C35 transcript in tumors was confirmed by Northern blot and real-time PCR. The C35 gene is located on chromosome 17q12, 505 nucleotides from the 3′ end of the ERBB2 oncogene, the antigenic target for trastuzumab (HerceptinTM) therapy. The chromosomal arrangement of the genes encoding C35 and ERBB2 is tail to tail. An open reading frame encodes a 12-kDa protein of unknown function. Immunohistochemical analysis detected robust and frequent expression of C35 protein, including 32% of grade 1 and 66% of grades 2 and 3 infiltrating ductal carcinomas of the breast (in contrast to 20% overexpressing HER-2/neu), 38% of infiltrating lobular carcinoma (typically HER-2/neu negative), as well as tumors arising in other tissues. C35 was not detected in 38 different normal human tissues, except Leydig cells in the testes and trace levels in a small percentage of normal breast tissue samples. The distinct and favorable expression profile of C35 spanning early through late stages of disease, including high frequency of overexpression in various breast carcinoma, abundant expression in distant metastases, and either absence or low level expression in normal human tissues, warrants further investigation of the relevance of C35 as a biomarker and/or a target for development of broadly applicable cancer-specific therapies. [Mol Cancer Ther 2006;5(11):2919–30]

Confocal Microscopy of Unfixed Breast Needle Core Biopsies: A Comparison to Fixed and Stained Sections

BackgroundNeedle core biopsy, often in conjunction with ultrasonic or stereotactic guided techniques, is frequently used to diagnose breast carcinoma in women. Confocal scanning laser microscopy (CSLM) is a technology that provides real-time digital images of tissues with cellular resolution. This paper reports the progress in developing techniques to rapidly screen needle core breast biopsy and surgical specimens at the point of care. CSLM requires minimal tissue processing and has the potential to reduce the time from excision to diagnosis. Following imaging, specimens can still be submitted for standard histopathological preparation.MethodsNeedle core breast specimens from 49 patients were imaged at the time of biopsy. These lesions had been characterized under the Breast Imaging Reporting And Data System (BI-RADS) as category 3, 4 or 5. The core biopsies were imaged with the CSLM before fixation. Samples were treated with 5% citric acid and glycerin USP to enhance nuclear visibility in the reflectance confocal images. Immediately following imaging, the specimens were fixed in buffered formalin and submitted for histological processing and pathological diagnosis. CSLM images were then compared to the standard histology.ResultsThe pathologic diagnoses by standard histology were 7 invasive ductal carcinomas, 2 invasive lobular carcinomas, 3 ductal carcinomas in-situ (CIS), 21 fibrocystic changes/proliferative conditions, 9 fibroadenomas, and 5 other/benign; two were excluded due to imaging difficulties. Morphologic and cellular features of benign and cancerous lesions were identified in the confocal images and were comparable to standard histologic sections of the same tissue.ConclusionCSLM is a technique with the potential to screen needle core biopsy specimens in real-time. The confocal images contained sufficient information to identify stromal reactions such as fibrosis and cellular proliferations such as intra-ductal and infiltrating carcinoma, and were comparable to standard histologic sections of the same tissue. Morphologic and cellular features of benign and cancerous lesions were identified in the confocal images. Additional studies are needed to 1.) establish correlation of the confocal and traditional histologic images for the various diseases of the breast; 2.) validate diagnostic use of CSLM and; 3.) further define features of borderline lesions such as well-differentiated ductal CIS vs. atypical hyperplasia.

Preliminary system characterization of flat-panel-detector-based cone-beam CT for breast imaging

Conventional film-screen mammography is the most effective tool for the early detection of breast cancer currently available. However, conventional mammography has relatively low sensitivity to detect small breast cancers (under several millimeters) owing to an overlap in the appearances of benign and malignant lesions, and surrounding structure. The limitations accompanying conventional mammography is to be addressed by incorporating a cone beam CT imaging technique with a recently developed flat panel detector. Computer simulation and preliminary studies have been performed to prove the feasibility of developing a flat panel detector-based cone beam CT breast imaging (FPD-CBCTBI) technique. A preliminary system characterization study of flat panel detector-based cone beam CT for breast imaging was performed to confirm the findings in the computer simulation and previous phantom studies using the current prototype cone beam CT scanner. The results indicate that the CBCTBI technique effectively removes structure overlap and significantly improves the detectability of small breast tumors. More importantly, the results also demonstrate CBCTBI offers good image quality with the radiation dose level less than or equal to that of conventional mammography. The results from this study suggest that FPD-CBCTBI is a potentially powerful breast-imaging tool.

Standardized Assessment of the HER2 Status in Breast Cancer by Immunohistochemistry

Immunohistochemistry (IHC) is widely used in surgical pathology, but it has been plagued by problems with reproducibility and lack of standardization resulting in poor concordance between laboratories. In particular, inaccuracy of routine human epidermal growth factor receptor 2 (HER2) testing in breast cancer patients has been a major issue. In 2006 this led the American Society of Clinical Oncologists (ASCO) and College of American Pathologists (CAP) to charge an expert panel with developing recommendations for HER2 testing. After subsequent publication and adoption of these guideline recommendations through dissemination of best practices, variation in clinical practice is expected to diminish and result in improved accuracy. In this article, we review the role of genomic HER2 alterations in the development and treatment of breast cancer, highlight the importance of accurate and reproducible HER2 testing, and discuss practical approaches to standardize HER2 testing by IHC. Pre-analytic and analytic variables are addressed, and a practical algorithm for test interpretation is introduced. * IHC : immunohistochemistry HER2 : human epidermal growth factor receptor 2 ASCO : American Society of Clinical Oncologists CAP : College of American Pathologists ER : estrogen receptor FISH : fluorescence in situ hybridization PR : progesterone receptor H&E : hematoxylin and eosin staining

The ubiquitin E3 ligase WWP1 decreases CXCL12-mediated MDA231 breast cancer cell migration and bone metastasis.

Advanced breast cancers preferentially metastasize to bone where cells in the bone microenvironment produce factors that enhance breast cancer cell homing and growth. Expression of the ubiquitin E3 ligase WWP1 is increased in some breast cancers, but its role in bone metastasis has not been investigated. Here, we studied the effects of WWP1 and itch, its closest family member, on breast cancer bone metastasis. First, we immunostained a multi-tumor tissue microarray and a breast cancer tissue microarray and demonstrated that WWP1 and ITCH are expressed in some of breast cancer cases. We then knocked down WWP1 or itch in MDA-MB-231 breast cancer cells using shRNA and inoculated these cells and control cells into the left ventricle of athymic nude mice. Radiographs showed that mice given shWWP1 cells had more osteolytic lesions than mice given control MDA-MB-231 cells. Histologic analysis confirmed osteolysis and showed significantly increased tumor area in bone marrow of the mice. WWP1 knockdown did not affect cell growth, survival or osteoclastogenic potential, but markedly increased cell migration toward a CXCL12 gradient in vitro. Furthermore, WWP1 knockdown significantly reduced CXCL12-induced CXCR4 lysosomal trafficking and degradation. In contrast, itch knockdown had no effect on MDA-MB-231 cell bone metastasis. Taken together, these findings demonstrate that WWP1 negatively regulates cell migration to CXCL12 by limiting CXCR4 degradation to promote breast cancer metastasis to bone and highlight the potential utility of WWP1 as a prognostic indicator for breast cancer bone metastasis.

Flat-panel detector-based cone-beam volume CT breast imaging: phantom and specimen study

Conventional film-screen mammography is the most effective tool for the early detection of breast cancer currently available. However, conventional mammography has relatively low sensitivity to detect small breast cancers (under several millimeters) owing to an overlap in the appearances of benign and malignant lesions, and surrounding structure. The limitations accompanying conventional mammography is to be addressed by incorporating a cone beam volume CT imaging technique with a recently developed flat panel detector. A computer simulation study has been performed to prove the feasibility of developing a flat panel detector-based cone beam volume CT breast imaging (FPD-CBVCTBI) technique. In this study, a phantom and specimen experiment is performed to confirm the findings in the computer simulation using the current prototype cone beam volume CT scanner. The results indicate that the CBVCTBI technique effectively removes structure overlap and significantly improves the detectability of small breast tumors. More importantly, the results also demonstrate the patient dose level required for FPD-based CBVCTBI to detect a small tumor (under 5 mm) and a small calcification is less than or equal to that of conventional mammography. The results from this study suggest that FPD-CBVCTBI is a potentially powerful breast-imaging tool.

Evaluation of flat panel detector cone beam CT breast imaging with different sizes of breast phantoms

The sensitivity to detect small breast cancers and the specificity of conventional mammography (CM) remain limited owing to an overlap in the appearances of lesions and surrounding structure. We propose to address the limitations accompanying CM using flat panel detector (FPD)-based cone beam CT breast imaging (CBCTBI). The purpose of the study is to determine optimal x-ray operation ranges for different sizes of normal breasts and corresponding glandular dose levels. The current CBCT prototype consists of a modified GE HighSpeed Advantage CT gantry, an x-ray tube, a Varian PaxScan 4030CB FPD, a CT table and a PC. Two uncompressed breast phantoms, with the diameters of 10.8 and 13.8 cm, consist of three inserts: a layer of silicone jell simulating a background structure, a lucite plate on which five simulated carcinomas are mounted, and a plate on which six calcifications are attached. With a single scan, 300 projections were acquired for all phantom scans. The optimal x-ray techniques for different phantom sizes were determined. The total mean glandular doses for different size phantoms were measured using a CT pencil ionization chamber. With the optimal x-ray techniques that result in the maximal dose efficiency for the different tissue thickness, the image quality with two different phantoms was evaluated. The results demonstrate that the CBCTBI can detect a few millimeter-size simulated carcinoma and ~ 0.2 mm calcification with clinically acceptable mean glandular doses for different size breasts.

The pen and the scalpel: effect of diffusion of information on nonclinical variations in surgical treatment.

Background:As information is disseminated about best practices, variations in patterns of care should diminish over time. Objective:To test the hypotheses that differences in rates of a surgical procedure are associated with type of insurance in an era of evolving practice guidelines and that insurance and site differences diminish with time as consensus guidelines disseminate among the medical community. Methods:We use lymph node dissection among women with ductal carcinoma in situ (DCIS) as an example of a procedure with uncertain benefit. Using a sample of 1051 women diagnosed from 1985 through 2000 at 2 geographic sites, we collected detailed demographic, clinical, pathologic, and treatment information through abstraction of multiple medical records. We specified multivariate logistic models with flexible functions of time and time interactions with insurance and treatment site to test hypotheses. Results:Lymph node dissection rates varied significantly according to site of treatment and insurance status after controlling for clinical, pathologic, treatment, and demographic characteristics. Rates of lymph node dissection decreased over time, and differences in lymph node dissection rates according to site and generosity of insurance were no longer significant by the end of the study period. Conclusions:We have demonstrated that rates of a discretionary surgical procedure differ according to nonclinical factors, such as treatment site and type of insurance, and that such unwarranted variation decreases over time with diminishing uncertainty and in an era of diffusion of clinical guidelines.

Real-time flat-panel detector-based cone beam volume CT breast imaging: phantom and specimen study

Conventional film-screen mammography is the most effective tool for the early detection of breast cancer currently available. However, conventional mammography has relatively low sensitivity to detect small breast cancers (under several millimeters) owing to an overlap in the appearances of benign and malignant lesions, and surrounding structure. The limitations accompanying conventional mammography is to be addressed by incorporating a cone beam volume CT imaging technique with a recently developed flat panel detector. Computer simulation and preliminary studies have been performed to prove the feasibility of developing a flat panel detector-based cone beam volume CT breast imaging (FPD-CBVCTBI) technique. In this study, a specimen experiment is performed to confirm the findings in the computer simulation and previous phantom studies using the current prototype cone beam volume CT scanner. The results indicate that the CBVCTBI technique effectively removes structure overlap and significantly improves the detectability of small breast tumors. More importantly, the results also demonstrate CBVCTBI offers good correlation with pathology images with the radiation dose level less than or equal to that of conventional mammography. The results from this study suggest that FPD-CBVCTBI is a potentially powerful breast-imaging tool.