Genomic characterization of HER2-positive breast cancer and response to neoadjuvant trastuzumab and chemotherapy-results from the ACOSOG Z1041 (Alliance) trial.

Background: HER2 (ERBB2) gene amplification and its corresponding overexpression are present in 15-30% of invasive breast cancers. While HER2-targeted agents are effective treatments, resistance remains a major cause of death. The American College of Surgeons Oncology Group Z1041 trial (NCT00513292) was designed to compare the pathologic complete response (pCR) rate of distinct regimens of neoadjuvant chemotherapy and trastuzumab, but ultimately identified no difference. Patients and methods: In supplement to tissues from 37 Z1041 cases, 11 similarly treated cases were obtained from a single institution study (NCT00353483). We have extracted genomic DNA from both pre-treatment tumor biopsies and blood of these 48 cases, and performed whole genome (WGS) and exome sequencing. Coincident with these efforts, we have generated RNA-seq profiles from 42 of the tumor biopsies. Among patients in this cohort, 24 (50%) achieved a pCR. Results: We have characterized the genomic landscape of HER2-positive breast cancer and investigated associations between genomic features and pCR. Cases assigned to the HER2-enriched subtype by RNA-seq analysis were more likely to achieve a pCR compared to the luminal, basal-like, or normal-like subtypes (19/27 versus 3/15; P = 0.0032). Mutational events led to the generation of putatively active neoantigens, but were overall not associated with pCR. ERBB2 and GRB7 were the genes most commonly observed in fusion events, and genomic copy number analysis of the ERBB2 locus indicated that cases with either no observable or low-level ERBB2 amplification were less likely to achieve a pCR (7/8 versus 17/40; P = 0.048). Moreover, among cases that achieved a pCR, tumors consistently expressed immune signatures that may contribute to therapeutic response. Conclusion: The identification of these features suggests that it may be possible to predict, at the time of diagnosis, those HER2-positive breast cancer patients who will not respond to treatment with chemotherapy and trastuzumab. ClinicalTrials.gov identifiers: NCT00513292, NCT00353483.

MRI vs. histologic measurement of breast cancer following chemotherapy: comparison with x-ray mammography and palpation.

Twenty consecutive patients with breast cancer were evaluated following chemotherapy using MRI to assess the size of cancer residua and compare these data with subsequent histologic measurements of the viable tumor. This retrospective study also involved assessment of the preoperative size of the malignancy as determined by physical exam and x-ray mammogram. These values were later compared with the histology. The tumor size correlation coefficient between MRI and pathologic analysis was the highest, at 0.93. Physical exam and x-ray mammography (available for 17 patients) produced correlation coefficients of 0.72 and 0.63, respectively, compared to histologic measurement. The accuracy of MRI did not vary with the size of cancer residua. MRI is an accurate method for preoperative assessment of breast cancer residua following chemotherapy. J. Magn. Reson. Imaging 2001;13:868-875.

Breast cancer: screening and early detection.

A comprehensive program for breast cancer screening and early detection has a number of components. First, an understanding of the magnitude of the breast cancer problem and the natural history of breast cancer is essential. Appropriate screening guidelines must be identified and adopted into practice. Screening recommendations must be promoted actively. While breast cancer screening guidelines can be applied broadly to the general population, patients who are at increased risk must be identified through formal risk assessment. For those at increased risk, earlier screening and prevention strategies can be recommended. Physicians must be prepared to manage the abnormalities identified in screening and make appropriate referrals for treatment. Finally, patients identified with cancer must be staged accurately, as this staging will determine the prognosis.

Breast cancer screening: success amid conflict.

In randomized trials, screening mammography has been shown to reduce breast cancer mortality. There has been controversy, however, about the segment of the population that should be screened and the frequency at which screening should occur. This article details these controversial issues. The benefits that have been realized from mammography screening in overall breast cancer mortality and diagnosis of earlier stage cancers are also reviewed.

Characterization of paired tumor and non-tumor cell lines established from patients with breast cancer.

The goal of our study was to develop a panel of tumor cell lines along with paired non-malignant cell lines or strains collected from breast cancers, predominantly primary tumors. From a total of 189 breast tumor samples consisting of 177 primary tumors and 12 metastatic tissues, we established 21 human breast tumor cell lines that included 18 cell lines derived from primary tumors and 3 derived from metastatic lesions. Cell lines included those from patients with germline BRCA1 and FHIT gene mutations and others with possible genetic predisposition. For 19 tumor cell lines, we also established one or more corresponding non-malignant cell strains or B lymphoblastoid (BL) lines, which included 16 BL lines and 7 breast epithelial (2) or stromal (5) cell strains. The present report describes clinical, pathological and molecular information regarding the normal and tumor tissue sources along with relevant personal information and familial medical history. Analysis of the breast tumor cell lines indicated that most of the cell lines had the following features: they were derived from large tumors with or without axillary node metastases; were aneuploid and exhibited a moderate to poorly differentiated phenotype; were estrogen receptor (ER)- and progesterone receptor (PR)-negative; and overexpressed p53 and HER2/neu proteins. Of 13 patients with primary breast cancers receiving curative intent mastectomies, 7 were dead after a mean period of 10 months. Our panel of paired tumor and non-malignant cell lines should provide important new reagents for breast cancer research.

American Cancer Society guidelines for the early detection of breast cancer: update 1997.

The American Cancer Society (ACS) convened a workshop in March 1997 to consider new scientific findings related to breast cancer screening and to determine whether these findings warrant a change in the existing ACS guidelines. The meeting was timed so that participants could benefit from new data related to screening women aged 40 to 49 years. A recommendation based on the new data and subsequently approved by the ACS Board of Directors is reported.

Controversies in breast cancer screening.

Because breast cancer is the most common cancer occurring in women in the United States, early detection of breast cancer through screening mammography, physician clinical examination, and breast self-examination has been recommended. However, despite admonitions to physicians and patients to be aggressive in their screening efforts, there has been continued controversy regarding appropriate guidelines for screening. In December 1993, the National Cancer Institute announced that it could no longer recommend routine mammographic screening for women age 40-49 years, which it had previously supported along with other medical organizations. This change in policy, along with data from the Canadian National Breast Screening Study showing an increased mortality rate for screened women, created confusion for physicians and patients alike. The controversy about screening guidelines has created many practical concerns for the physicians involved in the primary health care of women. In the current paper, the author discussed the development of screening guidelines and the current recommendations of various medical organizations and reviewed the data from studies supporting and challenging the current guidelines, with a focus on screening guidelines for women age 40-49 and the elderly. Recommendations are made for physicians on how to communicate with patients regarding screening controversies.

Breast cancer in a county hospital population: impact of breast screening on stage of presentation.

BACKGROUND: Indigent patients in a county hospital setting typically present with breast cancer at a later stage than do patients in the private sector. In the early 1980s, 50% of all breast cancers diagnosed in our county hospital were stages III and IV. This contrasted markedly with the findings of an American College of Surgeons study, which showed < 15% of breast cancers diagnosed as stages III and IV. METHODS: Recognizing this disparity, we instituted a breast screening project in the county teaching hospital targeted at women who routinely received medical care in the county hospital clinics. Between 1985 and 1992, 14,567 mammograms were performed. RESULTS: Two hundred eighty-nine breast biopsies were performed and 76 cancers were identified (26%). Ninety-five patients advised to have surgical consultation for biopsy declined further evaluation. The stage distribution of cancers diagnosed was as follows: stage 0, 20%; stage I, 43%; stage II, 28%; stage III, 8%; and stage IV, 1%. This compares favorably with National Cancer Data Base statistics for 1988. In contrast, symptomatic nonscreened patients diagnosed at the county hospital in 1992 presented at a significantly more advanced stage: stage 0, 1%; stage I, 14%; stage II, 45%; stage III, 26%; and stage IV, 13%. CONCLUSIONS: Mammographic screening has lowered the stage of cancers diagnosed in the screened indigent population. However, a significant percentage of patients are presenting to our hospital with stage III and IV disease. Problems identified in the screening project included noncompliance with recommendations for follow-up of abnormal studies and noncompliance with appointments. In order to broaden the impact of our breast screening project, we have instituted outreach programs with community-based clinics and the American Cancer Society.

Hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), and their receptors in human breast cells and tissues: alternative receptors.

We sought to determine whether the hepatocyte growth factor/scatter factor (HGF/SF)- and keratinocyte growth factor-receptor systems were expressed in normal breast cells, breast carcinoma cell lines, normal breast tissues, and breast cancer tissues. Reverse transcriptase-polymerase chain reaction and hot blotting were used to detect HGF, HGF/SF (met) receptor, KGF, and KGF receptor mRNAs in human mammary epithelial (HME) and stromal (HMS) cells. We also examined breast carcinoma (MDA-MB-157, SCC 38, and SCC 70) and spontaneously immortalized breast epithelial (HMT 3522) cell lines, as well as normal breast and breast carcinoma tissues. PCR products were also confirmed by nucleic acid sequencing. The effects of HGF and KGF, compared to EGF and heparin-binding EGF, on the proliferation of normal human mammary epithelial cells in serum-free defined medium was determined by cell counting. HGF and KGF mRNAs were detected in HMS cells, but not HME cells. KGF receptor mRNA was detected in HME cells, but not HMS cells. HGF/SF receptor mRNA was detected in both HME and HMS cells. mRNAs were also detected in normal breast and breast carcinoma tissues, as well as breast carcinoma and transformed breast epithelial cell lines. Alternative cDNA sequences that are predicted to code for a soluble KGF receptor and a membrane bound, truncated HGF/SF receptor were detected in breast epithelial cells and breast tissues. HGF and KGF maintained viability and stimulated proliferation of HME cells.

Multimodality treatment of locally advanced breast carcinoma.

Forty patients with 41 locally advanced breast lesions at stages IIIA and IIIB and the inflammatory stage were treated with combined-modality therapy from July 1980 to August 1985. Treatment included induction chemotherapy consisting of three cycles of fluorouracil, doxorubicin hydrochloride, and cyclophosphamide, followed by mastectomy in those patients whose lesions were operable (n = 28), and resumption of chemotherapy. Nine patients received postoperative radiation therapy. The mean follow-up was 34 months. Greater than 50% reduction in tumor size was achieved in 72% of patients after three cycles of chemotherapy. Overall, local control was achieved in 85% of patients with 59% survival and 53% disease-free survival, while 10% of patients developed local recurrences. Excluding lymphedema of the upper extremity (n = 2) and inflammatory carcinomas (n = 4), local control was achieved in 96% of patients, with 75% survival and 68% disease-free survival, while 4% of patients developed local recurrences. The rate of disease-free survival was 71% in patients with partial response to chemotherapy, contrasted with 43% in patients who did not respond or only minimally responded to chemotherapy. Actuarial five-year survival, based on life-table analysis, was calculated to be 46% for the group overall, 58% for the group excluding lymphedema of the upper extremity and inflammatory carcinoma, and 56% for the 28 patients undergoing mastectomy.

Detection of tumour associated antigen in eluates from protein A columns used for ex vivo immunoadsorption of plasma from melanoma patients by radioimmunoassay.

Tumour associated antigen (TAA) of defined specificity and anti-TAA antibodies were isolated by elution with 0.1 M glycine-HCl buffer (pH 3.5) and 2.5 M MgCl2 from non-viable Staphylococcus aureus used in ex vivo immunoadsorption of plasma from four melanoma patients. The anti-TAA antibody activity in the MgCl2 eluate was determined by its ability to bind a melanoma 125I-TAA. The melanoma 125I-TAA was isolated and purified from the spent culture medium of a human melanoma cell line. The activity and specificity of TAA in the glycine-HCl eluates were determined by competitive inhibition in a radioimmunoassay in which melanoma 125I-TAA and an allogeneic antiserum obtained from a melanoma patient were used as the reagents. Results indicated that 0.04-0.81% of the total protein contained in the glycine-HCl eluates was TAA. The proportion of TAA to total protein in these eluates varied from patient to patient and treatment to treatment. Inhibition by the glycine-HCl eluates in the competitive radioimmunoassay was dose-dependent. Similarly, binding of melanoma 125I-TAA in a direct radioimmunoassay decreased with decreasing amounts of the anti-TAA antibody fraction. Quantitative analysis revealed that the MgCl2 eluates contained anti-TAA protein at levels ranging from 0.15 to 5.78% of total protein. Because both TAA and anti-TAA activities were found in eluates from S. aureus (protein A positive) used for immunoadsorption of plasma from melanoma patients, and because melanoma 125I-TAA isolated and purified from a human melanoma cell line did not bind to protein A directly, the results indicated that TAAs immunologically similar to the melanoma TAA were circulating in the form of immune complexes in plasma of four patients with melanoma and that these complexes could be removed from plasma by ex vivo immunoadsorption.

Nature of antigens and antibodies in immune complexes isolated by staphylococcal protein A from plasma of melanoma patients.

Immune complexes (IC) were isolated from plasma of melanoma patients by absorption to staphylococcal protein A and subsequent elution with MgCl2. The isolated ICs were purified by precipitation with polyethylene glycol and sucrose density gradient ultracentrifugation after radioiodination with 125I. The purified ICs were dissociated and radiolabeled antigen/antibody components were separated by ultracentrifugation at low pH (2.6). Under these conditions, about 72% radioactivity of the purified IC remained in the light-density region as a wide band. After neutralization, 26%-60% radioactivity in the region of 5S sedimentation bound to immobilized autologous immunoglobulins, as opposed to a maximum of 23% to immobilized immunoglobulins from human normal serum. Significant levels (73%-77%) of radioactivity in 7S region bound to rabbit anti-human IgG immunobeads. Immunoprecipitation of the antigen fraction by allogeneic anti-melanoma and rabbit anti-melanoma antibodies followed by SDS-polyacrylamide gel electrophoresis revealed the presence of a fetal antigen (FA) and a melanoma tumor-associated antigen (TAA). In addition, the presence of auto-antigen(s) was indicated by using autologous antibody in immunoprecipitation. Immunoglobulins (IgG) isolated from purified IC bound to cultured melanoma, sarcoma, and normal fibroblasts, although the binding to sarcoma and normal fibroblasts could be inhibited by preincubation of isolated IgG with soluble FA but not with soluble melanoma TAA. Thus, results of this investigation provide evidence that circulating IC in melanoma patients are composed of at least IgG and different antigens, and some of these antigens are produced by their tumor.