bluuebunny/crossref_metadata_2025 huggingface dataset:
Physical activity and risk of postmenopausal breast cancer defined by hormone receptor status and histology: A large prospective cohort study with 18 years of follow up
Authors: Bardia A. , Wang A. H. , Hartmann L. C. , Olson J. E. , Vachon C. M. , Vierkant R. A. , Cerhan J. R.
Published: March 2020

1002

Background: Physical activity is a modifiable breast cancer risk factor, perhaps mediating risk reduction through regulation of estrogen metabolism. Evidence regarding effect of physical activity is conflicting partly because breast cancer is a heterogenous constellation of different tumor subtypes with differing etiologies. No prospective study has examined the relationship between physical activity and breast cancer incidence based on ER/PR status or histological subtype. Objective: Examine effect of physical activity on breast cancer incidence based on ER/PR status and histological subtypes of breast cancer. Methods: The Iowa Women’s Health Study is a prospective cohort study of postmenopausal women (N=41,837). Physical activity was self-reported on baseline questionnaire, and three levels (high, medium and low) were defined. Breast cancer incidence, histologic subtype and ER/PR status, through 18 years of follow-up, were ascertained by linkage with the Iowa SEER Cancer Registry. Cox proportional hazards models were used to estimate multivariate relative risks (RRs) and 95% confidence intervals (CIs) of breast cancer, adjusting for other breast cancer risk factors. Results: During 554,819 person-years of follow-up, 2548 incident cases of breast cancer were observed. High physical activity was associated with decreased risk for breast cancer (RR 0.91, 95 % CI 0.81–1.01) compared to low activity. The protective effect was most marked in ER+/PR− (RR 0.66, CI 0.46–0.94), intermediate in ER−/PR− (RR 0.80, CI 0.56–1.15), weakest in ER+/PR+ (RR 0.94, CI 0.81–1.08), and elevated in ER-/PR+ (RR 1.42, CI 0.67–3.01) tumors. Higher physical activity was also associated with a decreased risk of invasive ductal/lobular carcinoma (RR 0.90, CI 0.80–1.02), but not with invasive breast cancer with a favorable histology (RR 1.19, CI 0.78–1.81). Conclusions: Higher physical activity was associated with a 10% decreased risk of breast cancer. Unexpectedly, risk reduction was most marked in PR- tumors, particularly ER+/PR-, and the more aggressive histologic forms. Further studies are needed to confirm these findings, and also evaluate other risk factors based on ER/PR status and histological subtypes.

No significant financial relationships to disclose.
Histopathological features of breast cancers in women with germline TP53 mutations
Authors: Masciari S. , Kandel M. , Digianni L. , Dillon D. , Li F. , Garber J.
Published: March 2020

10031

Background: Breast cancers (BR) in women with germline BRCA1 mutations are usually high-grade ductal carcinomas; 90% are negative for estrogen receptor (ER), progesterone receptor (PR) and HER2. No characteristic phenotype has been found for BRCA2-associated tumors. For women with germline TP53 mutations (Li Fraumeni Syndrome), breast cancer is the most common tumor; the risk of BR is 49% by age 60, with significant risk before age 40. The histopathological profile of BR in women with germline TP53 mutations has not been characterized. Methods: We identified BR from 17 female TP53 mutation carriers from the LFS family registry at Dana Farber Cancer Institute on which at least ER and PR data were available. Information on histology, hormone receptor and HER2 status by immunohistochemistry (IHC) was collected from medical records. Histology was not centrally reviewed. Results: 14 invasive ductal carcinoma (IDC) and 4 DCIS were identified in 17 women (1 woman had both) (see Table). The median age at diagnosis of invasive BR was 30 years (range 24–48), for DCIS it was 37 years (range 22–40). Of the 14 IDC, 11 (79%) were high grade (G3), 10 (71%) were ER positive, 7 (50%) PR positive, 6 PR negative and 1 PR unknown. Among 9 tumors with available HER2 status, 6 (67%) were positive. Both ER+/PR+ and ER-/PR- DCIS were observed. Conclusions: In young women with TP53 germline mutations, BR are more likely to be high grade and ER positive with no other distinct pattern in contrast to BRCA1-associated tumors. Findings of DCIS alone suggest potential value for mammographic screening. Confirmation of these results in a larger series will be important.

[Table: see text]

No significant financial relationships to disclose.
The ING tumor suppressor genes and their specific role in the pathogenesis of renal cell carcinoma
Authors: Nichiporuk E. , Lebedeva T. , Gasser M. , Hillig F. , Lutz J. , Grimm M. , Kneitz B. , Heemann U. , Riedmiller H. , Waaga-Gasser A.
Published: March 2020

10045

Background: The inhibitor of growth (ING) family of tumor suppressor genes is involved in cell cycle arrest, regulation of gene transcription as well as DNA repair. p33ING1b plays an important role in the pathogenesis of certain carcinomas by modulation of p53. We analyzed p33ING1b and p29ING4 gene expression together with specific immune responses in patients with renal cell carcinoma (n=50) at different tumor stages. Methods: Peripheral blood lymphocytes (PBMCs) from patients (Robson stage I-IV) were stimulated with pools of synthetic overlapping peptides of the p33ING1b or p29ING4 sequences encompassing the full length sequence of these two genes. PBMCs and tumor specimens were further characterized (ELISPOT, FACS, immunohistology, Real Time PCR). Results: T cells from stage I/II patients expressed higher IL-10 (n=5) than IFN-γ (n=5) levels in response to p29ING4 peptides. However, distinct residues of peptides were found that induced a Th2 type response (IL 10, n=5) in stage III/IV patients. Interestingly, distinct residues induced a Th1 (IFN-γ, n=5) response in the latter patients. Lymphocytes stimulated with p33ING1b peptide pools expressed IFN-γ as well as IL-10, independently from the tumor stage. Remarkably, immunohistochemical staining as well as Real Time PCR analysis of tumor specimens revealed higher numbers of CD4/CD8, CD4/CD25, CD4/Foxp3, CD4/CTLA-4, and NK cells as well as IL-10, IFN-γ, and Annexin V expression at the tumor site of stage I/II patients than later tumor stages. However, stronger staining and gene expression of p33ING1b as well as p29ING4 together with a reduced staining and expression of p53 was observed in stage III/IV patients. A correlation between the stage and the grading of the tumor was not present. Conclusions: In order to exert its function as a growth arrest and apoptosis inducing protein, p53 needs to interact with other tumor suppressor genes like the ING gene family. Subsequently, the loss of ING function may be a potential mechanism for the inactivation of p53 function in renal cell carcinoma. The results of this study may provide the basis for immune therapeutical strategies (induction of apoptosis or of a Th1 response using a vaccination protocol in particular with p29ING4 in the early stage of the disease) in renal cell carcinoma.

No significant financial relationships to disclose.
Immunophenotyping of solid tumors, a new approach for molecular profiling solid tumors
Authors: Nunez R. , Magenau J. , Zamorano S. , Lostumbo A. , Mehta D.
Published: March 2020

10057

Methods of determining prognosis in breast, ovary, and colon cancer includes detection of molecular markers such as the estrogen receptor (ER), progesterone receptor (PR), and HER-2/neu. These markers are routinely assessed via immunohistochemistry (IHC) and Fluorescent In-situ hybridization (FISH). Flow cytometry (FC) has not yet been established for routine detection of these markers in tumors. We performed FC on cells from fourteen ovary, five breast, and four colon cancer tumors. FC was used to detect the presence of ER, PR, HER-2/neu, Epidermal Growth Factor Receptor (EGFR), E-cadherin, RAS, ERK and TUJ1. Cells obtained directly from tumoral tissue were simultaneous analysized by cell cycle stage (G0, G2M, and > G2M) and presence of antigenic markers. In ovarian tumors, 7 of 14 samples demonstrated >50% of cells with her-2/neu expression in aneuploid fractions (>G2M), 9 of 14 in G2M phase, and only 2 of 14 in G0 phase. With respect to ER, 7 of 14 ovarian tumors exhibited >19% of cells in G2M and >G2M fractions, while only 1 of 14 samples had >19% at G0. The PR receptor was expressed in >19% of cells in 8 of 14 (>G2M), 7 of 14 (G2M), and 2 of 14 (G0). In breast tumors, 4 of 5 tumors demonstrated >50% expression of HER-2/neu in G2M and >G2M cell phases, and only 2 of 5 demonstrated >50% her2neu expression in G0. Three of five tumors had >19% cellular expression of ER in the >G2M and G2M phases, while only one had >19% in G0. With PR, 3 of 5 breast tumors had >19% cellular expression in G0, G2M, and >G2M phases. In colon tumors, 2 of 4 had >50% cellular expression of her2neu in >G2M fractions, 4 of 4 in G2M, and only 1 of 4 in G0. Interestingly, cell fractions containing G2M and > G2M DNA had > 50% cellular expression for RAS and ERK in 2 of 4 colon tumors. No colon tumors expressed RAS and ERK significantly in G0 cells. We found that ER, PR, and HER-2/neu marker expression were consistent with established expression patterns. Additionally, these data point to specific expression patterns that are associated with the cell cycle stage. In particular, we note high expression levels of Her-2/nue, ER, and PR in aneuploid (>G2M) cell populations. These results suggest that patterns of differential expression can be readily assessed in tumor tissues using flow cytometry, and has the potential for broad application to the study of solid tumors.

No significant financial relationships to disclose.
Active c-Src has prognostic and therapeutic value in ER-negative breast cancer
Authors: Arnaout A. , Yang L. , Holloway C. , Steven N. , Sun P. , Kahn H.
Published: March 2020

10063

Background: Approximately 33% of newly diagnosed breast cancers lack ER and these tend to have a worse prognosis as compared to ER-positive breast cancers. Therapeutic options are limited as they are not responsive to antihormonal therapy and often develop resistance to chemotherapies. We have recently shown that activation of the nonreceptor tyrosine kinase protein c-Src leads to the accelerated ER degradation in ER-negative breast cancers. This study performs an immunohistochemical analysis of activated c-Src in a large cohort of primary human breast carcinomas to a) assess its prognostic significance and b) correlate its relationship to the ER status of breast cancers. Methods: A total of 916 patients with breast cancer diagnosed between 1987 and 1997 had clinicopathological data and paraffin-embedded tumor tissues for the study. Tissue microarrays were constructed. A four point scoring system based on immunostaining intensity was used to grade the levels of active phosphorylated c-Src. Grading was done by one pathologist. Statistical analysis was used to assess the prognostic significance of activated c-Src and its relationship to other prognostic variables. Results: Median follow-up was 7.31 years. Active c-Src grade was inversely correlated with ER status (p=0.004) and predicted for treatment with chemotherapy (p=0.002) and lack of treatment with Tamoxifen (p=0.007). Patients with greater levels of c-Src tended to be younger (p=0.004) and had higher Bloom Richardson scores for their tumors (p=0.004). Higher levels of c-Src also predicted for for shorter timing to distant recurrence (p=0.01) and shorter timing to death (p=0.04). There was a trend towards a shorter timing to regional recurrence with higher levels of c-Src but the relationship was not statistically significant (p=0.08). Conclusion: This study supports the hypothesis that the presence of active, phosphorylated c-Src contributes to the development of ER-negative status in breast cancers. The presence of c-Src also is associated with other poor prognostic factors and contributes to a worse prognostic outcome. This study suggests that c-Src inhibitors may be a novel therapeutic strategy for the treatment of ER-negative breast cancers.

No significant financial relationships to disclose.
Use of denaturing high performance liquid chromatography (dHPLC) for detection of EGFR mutations in patients with NSCLC
Authors: Agulnik J. S. , Cohen V. , Jarry J. , Batist G. , Small D. , Kreisman H. , Tejada N. A. , Chong G. , Miller W. H.
Published: March 2020

10068

Background: Somatic mutations of the epidermal growth factor receptor (EGFR) gene may predict responsiveness to tyrosine kinase inhibitors in patients with non-small cell lung cancer (NSCLC). These mutations are commonly identified using DNA sequencing methods. Although considered the gold standard, this approach is expensive and time-consuming. In addition, a high ratio of tumour to normal tissue DNA is required for optimal results which is not often available in biopsies obtained from these patients. The primary objective of this study was to develop a rapid and sensitive method for screening of EGFR mutations. Materials and Methods: Clinical specimens from 110 NSCLC patients were analysed for EGFR mutations in exons 19 and 21. After DNA extraction and PCR, both dHPLC and direct sequencing were performed and results were compared. Results: Sequencing revealed a total of 7 (6%) mutations: 4 missense mutations in exon 21 and 3 deletion mutations in exon 19. dHPLC showed the presence of genomic alterations in 18 (16%) samples, including the 7 identified by sequencing plus 11 additional samples (10 in exon 19 and one in exon 21). dHPLC fractions were isolated, reamplified, and sequenced to confirm these results. In serial dilution studies, dHPLC was able to detect mutations in samples containing as little as 10% mutated DNA whereas direct sequencing required at least 30%. Conclusions: dHPLC is an efficient and accurate, as well as a a more sensitive method for screening of genomic alterations in exons 19 and 21 of the EGFR gene compared to direct sequencing. This data suggests that dHPLC should be implemented as a screening tool for detection of EGFR mutations.

No significant financial relationships to disclose.
Study of BRAF mutations in melanoma and nevi from patients with germline mutation in the CDKN2A gene
Authors: Robert C. , Rouiller J. , Kannengiesser C. , Chompret A. , Avril M. , Dessen P. , Michiels S. , Sarasin A. , Bressac de Paillerets B. , Spatz A.
Published: March 2020

10075

Background: Germline mutations in the CDKN2A tumor suppressor gene is a rare condition associated with a high risk of melanoma. Somatic activating mutations of the BRAF gene are frequently observed in cutaneous melanoma (40–60%) but their occurrence vary according to melanoma subtypes (cutaneous, vs mucosal or uveal) and are less frequent in melanoma located on chronically sun-exposed skin. It is thought that melanomas occurring in patients carrier of a CDKN2A germline mutation are associated a second inactivating genetic event in the tumor, facilitating cellular transformation by loss of function this crucial gatekeeper of the G1-S checkpoint. However, the potential role of the MAP-kinase pathway, and more precisely, the involvement of BRAF activation, in this particular population of patients, are still unknown. Thus, our objective was to evaluate the frequency of BRAF somatic mutations in melanoma and nevi developed by patients carrier of a CDKN2A germline mutation. Methods: DNA was extracted from paraffin-embedded tissues of 36 primary melanomas, 9 metastases and 20 nevi from 31 patients with CDKN2A germline mutation. Ten sporadic melanoma from patients harbouring no CDKN2A mutation were also studied as a control population. BRAF mutations were screened by direct sequencing of exon 11 and 15. Results: BRAF mutations (V599E) were found with a significantly lower frequency of 14% (5/36) in melanomas from patients with proven CDKN2A germline mutation, as compared to a frequency of 55% (5/9) in patients with no genetic predisposition (two-sided Fishers exact test, p=0.02). Frequencies of BRAF mutations in metastases and nevi from patients with CDKN2A germline mutation were 33%(3/9) and 10% (2/20) respectively. Conclusions: BRAF mutations seem to be less frequent in melanoma from patients with familial predisposition harbouring CDKN2A germ line mutation than in sporadic melanoma. These results suggest that CDKN2A loss of function gives rise to tumorigenic pathways where genetic events distinct from BRAF activating mutation can induce cell transformation. Additional biological studies are performed on this rare population of melanoma samples in order to better characterize the specificity of cellular transformation in the context of CDKN2A germ line mutation.

No significant financial relationships to disclose.
Recovery of polyclonal immunoglobulin serum levels to normal levels after autologous stem cell transplantation predicts disease free survival in patients with multiple myeloma
Authors: Joao C. , Geyer S. , Markovic S. , Gertz M. , Lacy M. , Dispenzieri A. , Kumar S. , Hayman S. , Gastineau D. , Porrata L.
Published: March 2020

10088

Background: The ability of the immune system to recover its components after autologous stem cell transplantation (ASCT) is known to predict survival in patients with different types of neoplasms. Also, our previous work showed that polyclonal immunoglobulin can be used to improve T cell diversity and, consequently, T cell function in animal models. The aim of this retrospective study was to understand if the recovery of the polyclonal compartment of immunoglobulin in patients with multiple myeloma (MM) at day 100 (± 40d) after ASCT is a predictor factor for survival in MM patients who were subject to ASCT. Methods: From294 patients with IgG-MM subject to ASCT at the Mayo Clinic, Rochester, MN since October, 1993 until June, 2004, 143 patients were studied. The rational to select this group of patients was the fact that they had levels of serum IgM collected at day 100 ± 40 after ASCT recorded. Progression free survival (PFS) was assessed from the day of the transplant. PFS was studied using Kaplan-Meier curves and the univariate and multivariate analysis were done using age-adjusted proportional Cox. The prognostic variables in the multivariate model were: recovered of IgM at day 100, B2 microglobulin, age, conditioning regimen and disease status at the time of transplant. Results: There were 59 females in the study. The median age of the cohort was 58 y (33–75). The median follow up of the study was 26 mo (range, 2-111). PFS was significantly higher in the group who recovered normal serum levels of IgM at D100 compared to the pts who did not. This corresponds to a survival that is 1.5 longer in the group of patients who recovered normal levels of IgM at day100 after ASCT versus the patients who did not (median: 34 to 23 mo, respectively; p=0.05). By univariate analysis, those who did not recover normal serum levels polyclonal immunoglobulin by day 100 after ASCT had a risk of relapse 65% higher than the risk of the patients who recovered. Multivariate analysis suggests that recover the normal levels of polyclonal immunoglobulin after ASCT is an independent prognostic factor for PFS for MM patients after ASCT. Conclusions: These results suggest that polyclonal immunoglobulin is a major factor promoting immunity after ASCT.

No significant financial relationships to disclose.
Identification of ABCA5 as a tissue and urine diagnostic marker for prostatic intraepithelial epithelial or PIN
Authors: Stearns M. E. , Hu Y. , Wang M. , Veverka K.
Published: March 2020

10089

Background: Bostwick and colleagues have suggested that prostatic intraepithelial neoplasia or PIN is the precursor of prostate cancer and that a significant percentage of men with PIN develop prostate cancer (Sakr, 1994; Bostwick, 1995b; Arakawa, 1995; Qian, 1995; Qian, 1997). Currently there are no diagnostic markers for PIN. Methods and Results: Recently, we have developed ‘DNA-protein’ binding assays for the identification of novel transcriptional regulatory proteins associated with PIN glands. Electrophoretic mobility assays (EMSAs) revealed that 1 (of 4096 sequences screened) DNA sequences specifically bound a 160 Kda protein over-expressed in PIN and not found in BPH, SV or PCA glands (n=11). The gene encoding for the 160 Kda protein was cloned from a lamda ZAP expression library by ‘in situ’ hybridization assays with the 32P labeled DNA sequence. Sequencing revealed that the gene encoding for the 160 Kda protein was ABCA5, a member of the superfamily of ATP-binding cassette (ABC) transporters of multi-drug resistant genes. RT-PCR assays indicated the mRNA was over-expressed in primary PIN cell cultures and by PIN tissue, but it was not present in BPH, or SV tissue. Rabbit polyclonal antibodies were raised against a n-terminal peptide (20 b) plus a c-terminal peptide (20b) and found to specifically label the cell junctions of 3% glutaraldehyde fixed cultured cells and to label putative adult cancer stem cells in sections of PIN tissue. ‘Blinded’ ELISA studies with urine specimens from biopsed patients showed that ABCA5 was over-expressed urine from patients with PIN (n=∼20), faintly expressed in PIN + PCA patients (n=8), and not expressed in BPH (n=10) or normal patients (n=20). Conclusions: The data suggest that the expression of ABCA5 is over-expressed by PIN and is a urine diagnostic marker for detection of the disease (>95% COI). Supported by NIH-NCI grant CA90397 to mes and by GTx.

[Table: see text]
The influence of efaproxiral on breast cancer xenograft hypoxia and HIF-1-alpha expression
Authors: Kavanagh B. D. , Steffen R. P. , Frederick B. , Solomon B. , Chan D.
Published: March 2020

10102

Background: Efaproxiral (E), a synthetic allosteric modifier of hemoglobin, has demonstrated clinical safety and efficacy as a radiosensitizer in patients with brain metastases from breast cancer. Although shown to enhance oxygenation in a murine mammary tumor, efaproxiral has not previously been tested in a human breast cancer xenograft. We studied changes in tumor oxygenation in a human xenograft and evaluated whether efaproxiral induces cell signaling events of potential therapeutic value. Methods: MDA-MB-468 breast cancer cells in matrigel were injected into the flank of nude mice. After tumors grew to 1–1.5 cm, animals were subjected to 1 of 3 treatments: ip saline +room air breathing (RA), ip saline + 50% oxygen breathing (O2), or ip efaproxiral (300 mg/kg) + 50% oxygen breathing (E+O2). Twenty minutes later the hypoxia marker, pimonidazole (pimo), was given, and 70 minutes later tumors were harvested for immunohistochemical study of hypoxia and hypoxia-inducible factor 1-alpha (HIF1-α) and RNA extraction to identify early changes in gene expression. Image analysis software was used to quantify observations. Results: Tumor hypoxia and HIF1-α staining were significantly decreased by efaproxiral (Table). HIF1-α staining did not entirely colocalize with pimo, implying different oxygen tension levels for HIF1-α ubiquitination and pimo reduction. Gene arrays indicated that after E+O2, expression of the hypoxia-induced DR1 transcription repressor was reduced compared with O2. Conclusions: The combination of E+O2 reduced hypoxia and HIF1-α expression in MDA-MB-468 human breast tumors in vivo, and an early effect on gene expression was reduced DR1. The results demonstrate an efaproxiral-mediated enhanced oxygenation of human hypoxic breast cancer. Furthermore, the efaproxiral-mediated down-regulation of HIF1-α suggests possible new opportunities in the clinical application of efaproxiral, notably as an adjuvant to systemic agents for which HIF1-α-mediated resistance limits efficacy.

[Table: see text]

[Table: see text]
Clinical factors that affect breast cancer risk reduction decisions in high risk women tested for the BRCA1/2 genetic mutation
Authors: Uyei A. R. , Broglio K. R. , Solomon T. L. , Vogel K. J. , Amos C. I. , Lu K. , Meric-Bernstam F. , Strong L. , Arun B.
Published: March 2020

1013

Background: Women with an increased risk for breast cancer have many risk reduction options including: prophylactic mastectomy, prophylactic oophorectomy, chemoprevention, and screening. Women without breast cancer make such decisions in a purely preventive setting and factors that affect their decisions are unclear. Method: We performed an IRB approved retrospective review of the medical records on women who underwent BRCA testing. We evaluated the women without a history of breast cancer to assess clinical characteristics and their relation to decision making. The risk reduction categories analyzed were: prophylactic mastectomy, prophylactic oophorectomy, tamoxifen, increased surveillance with MRI, and standard screening (clinical breast exam and mammography). Patient characteristics were tabulated by clinical decision group and the chi-square test or Fisher’s exact test was used. Results: From 2001, 627 patients have undergone genetic testing. 202 of these women did not have a history of breast cancer among whom 58 were mutation carriers. Most patients chose standard screening (47%) or increased surveillance (38%). 4% chose tamoxifen, 7% chose prophylactic mastectomy, 3% chose both prophylactic mastectomy and oophorectomy, and 5% chose oophorectomy. The tamoxifen group was too small to do further analysis. Increased surveillance did not show any significant association with any of the clinical factors that we evaluated. The majority of women who chose standard screening had a personal history of ovarian cancer (p<0.0001) and had no family history of ovarian cancer (p=0.02). Prophylactic surgeries were significantly associated with positive BRCA status (p=0.01). Women with a family history of ovarian cancer tended to have prophylactic surgery (p=0.02). Women who had DCIS or a breast biopsy tended to have prophylactic mastectomies (p=0.0001 and p<0.001 respectively). Conclusion: In breast cancer free women, BRCA status, family history of ovarian cancer, DCIS, and breast biopsy were associated with prophylactic surgeries. Having ovarian cancer or no family history of ovarian cancer were associated with standard screening. We are performing a questionnaire study to determine the reasons behind these women’s choices.

No significant financial relationships to disclose.
Neoadjuvant chemotherapy (NCT) in 710 patients for operable breast cancer: Comparison of 2 pathological classifications
Authors: Penault-Llorca F. , Abrial C. , Raoelfils I. , Amat S. , Mouret-Reynier M. , Leheurteur M. , Durando X. , Dauplat J. , Curé H. , Chollet P.
Published: March 2020

10502

Background: In a database harbouring the patients of several prospective phase II neoadjuvant trials (avcf, avcfm, fec 50–100, nem, net, tncf, taxotère/tncf, taxotere alone), clinical and pathological responses rates were studied in 710 women with stage II-III operable breast cancer treated 1982–2004. Pathological review was conducted to assess the residual disease in breast and nodes according to the 2 most used classifications in Europe: CHEVALLIER’s and SATALOFF’s (Am J Clin Oncol 1993; J Am Coll Surg 1995). Methods: Median age of the patients was 49.5 years [26–81]. Median diameter of the invasive tumour was 40 mm [10–130]. 555 (78%) patients had a canalar, 90 (13%) a lobular, 18 (2.5%) a mixed or invasive carcinoma, 36 (5%) neoplasic cells only and 11 (1.5%) another carcinoma. 25.6% of the tumours were grade III SBR, 28.3% grade 4 or 5 MSBR. The median number of NCT courses was 6 [1–9] followed by a surgery for 92%, a radiotherapy for 95%, an adjuvant chemotherapy (17%) and/or a hormonotherapy (52%). A breast evaluation was realized before, during and after NCT (Amat et al, Breast Cancer Res Treat 2005). Results: Intent to treat (n = 710), overall response rate was 68% (16% complete). The complete pathological response (pCR) rate was 14.18% according to Chevallier’s and 22.50% according to Sataloff’s classification. On 656 patients operated, 470 (72%) had a conservative surgery. On 520 patients with an axillary dissection, 272 (52.3%) had involved nodes (median number:1 [0–20]). After a median follow-up of 93 months, DFS and actuarial survival at 120 months were 54.9% and 66.5%, respectively.Chevallier’s classification level 1 (pCR in breast and axilla) was the most predictive of a good DFS, with a plateau appearance near 80%. Chevallier’s classification level 2 (in situ only) and Sataloff grade A (pCR and isolated tumor cells) yielded a lesser DFS (p < 0.01). With a long follow-up, a complete pCR is the most favourable prognostic factor, followed by in situ only or isolated tumor cells, then residual tumor. Conclusions: Sataloff’s gives higher pCR figures than Chevallier’s without evidence of superior predictive value. The classification used is important to predict outcome after NCT.

No significant financial relationships to disclose.
Molecular risk estimation and adjuvant chemotherapy in node-negative breast cancer patients—A status report of the prospective clinical trial NNBC 3-Europe
Authors: Paepke D. M. , Herbst F. , Annecke K. , Gaskill N. , Sweep F. , Meisner C. , Schmitt M. , Jaenicke F. , Harbeck N. , Thomssen C.
Published: March 2020

10503

Background: Feasibility of risk assessment in node negative breast cancer using the invasion markers urokinase-type plasminogen activator uPA and its inhibitor PAI-1 has been demonstrated in several prospective and retrospective studies and in a large meta analysis. Patients with low uPA and/or PAI-1 concentrations in tumor tissue levels have an excellent 5-year overall survival (>95%) even without any adjuvant therapy. Use of these molecular markers may spare adjuvant chemotherapy in approximately one half of node-negative breast cancer patients. Particulary, patients with an intermediate grade tumor can easily be differentiated in low and high risk. In addition, patients with high uPA and/or PAI-1 level seem to benefit from adjuvant chemotherapy. The NNBC-3-Europe trial seeks to answer two questions: 1. Is risk assessment by molecular markers uPA/PAI-1 superior to that by clinico-pathological factors regarding identification of low-risk patients? 2. Is adjuvant chemotherapy with anthracycline-taxane sequence (FEC-Docetaxel) superior to standard FEC in high-risk patients? Methods: In the NNBC-3-Europe trial, participating centres opt to either perform risk estimation by clinico-pathological factors or by the uPA and PAI-1 concentration in tumor tissue biopsies. Low-risk patients will be observed without adjuvant chemotherapy. High-risk patients are randomised based on the kind of adjuvant chemotherapy. Patients with steroid hormone receptor positive tumors receive adequate endocrine therapy. Results: Of the first 506 patients in the study arm with molecular risk assessment, 62 had a grade 1 tumors, and 184 had grade 3 tumors. Among grade 2 tumors (n = 260), 87 had low uPA/PAI-1. 173 patients presented with high levels. About 30% of the patients were allocated to the low-risk group using uPA/PAI-1. Conclusions: An adjuvant chemotherapy trial based on uPA/PAI-1 determination in the primary tumor is feasible in a multicentre setting. Applying these molecular markers for risk assessment, at this stage of the study, adjuvant chemotherapy could be spared in almost one third of N0 patients. It is performed in association with the EORTC PathoBiology Group Receptor und the German AGO Breast Group.

[Table: see text]
Letrozole (Femara) causes potent suppression of breast cancer tissue estrogen levels in the neoadjuvant setting
Authors: Geisler J. , Helle H. , Ekse D. , Duong N. K. , Evans D. , Lønning P. E.
Published: March 2020

10532

Background: Aromatase inhibitors of the third generation like letrozole, anastrozole and exemestane are known to suppress plasma estrogens by 90–99% in postmenopausal women. However, little is known about their influence on tissue estrogen levels. Methods: We investigated the effect of neoadjuvant treatment with letrozole (2.5 mg o.d.) given for 16 weeks to patients with locally advanced breast cancers on tissue levels of estradiol (E2), estrone (E1) and estrone sulfate (E1S) measured with a previously published, highly sensitive, HPLC-RIA method (J. Steroid. Biochem. Mol. Biol. 72, 259–264, 2000). All patients had either estrogen receptor (ER) and/or progesterone receptor (PGR) positive tumors (at least one of the receptors expressed in > 50% of the tumor cells). Results: Mean tissue levels of E2, E1 and E1S were found to be 475.8, 272.4, and 160.5 fmol/g tissue at baseline (geometric mean values). Following 16 weeks of treatment with letrozole, these levels fell to 11.3, 18.2, and 16.0 fmol/g corresponding to a suppression of tissue levels of E2, E1 and E1S by 97.6%, 90.7%, and 90.1%, respectively. All plasma levels of E2, E1 and E1S were in the normal range expected for postmenopausal women prior to treatment with letrozole. The individual plasma estrogen fractions were found suppressed by 96.6%, 99.1% and 99.5%, respectively, during treatment with letrozole. Conclusions: Treatment with neoadjuvant letrozole 2.5 mg o.d. potently suppressed tissue estrogen levels in postmenopausal breast cancer patients by 90.1 to 97.6%, surpassing the results previously reported for anastrozole in a similar setting (Clin. Cancer Res. 7, 1230–1236, 2001). While a significant superiority of letrozole versus anastrozole concerning total body aromatase inhibition and plasma estrogen suppression has previously been recorded (J. Clin. Oncol. 20 (3), 751–757, 2002), we are now able to extend this observation to tissue estrogen levels as well. Letrozole (2.5 mg o.d.) seems superior to anastrozole 1 mg daily with respect to plasma as well as tissue estrogen suppression, advocating head to head comparison of these compounds in early breast cancer.

[Table: see text]
Changes to the hormone receptor expression in patients treated with either tamoxifen or aromatase inhibitors as endocrine therapy for breast cancer
Authors: Djahansouzi S. , Rostock V. , Niederacher D. , Rath W. , Bender H. , Hanstein B.
Published: March 2020

10543

Background: Endocrine therapy (ET) of patients with hormone receptor (HR)-positive breast cancer (BC) is thought to adversely affect the HR expression of the tumor with time. In order to evaluate this, a study was undertaken comparing the HR status prior to adjuvant Tamoxifen (Tam) therapy and after progression in estrogen receptor (ER)-positive BC patients. Similarly, ET with aromatase inhibitors (AI) was investigated by comparing the HR status prior to AI therapy and after progression in ER-positive metastatic BC patients. Methods: A retrospective analysis of all stage I-III ER-positive BC patients who presented to the department between 1999 to 2004 and received adjuvant Tam for ≥ 6 month and had the tumor relapse (local recurrence or distant metastasis) confirmed by tumor biopsy. All patients were evaluated for ER and progesterone receptor (PR) status prior to ET as well as after relapse using immunohistochemistry. Similarly, stage IV ER-positive metastatic BC patients who received AI for ≥ 3 month with tumor biopsy prior and after AI therapy, were also analyzed in regards to their HR expression. Results: 44 eligible ER-positive BC patients constituted the Tam group. After relapse, 55% of the patients remained ER-positive, while 45% lost their positive ER status. For PR, 34 out of 44 patients were initially receptor positive (77%). Similarly, 15 out of 34 patients retained a positive PR expression (44%) and the 10 PR-negative patients showed no change in receptor expression following Tam. 5 eligible patients with ER-positive metastatic BC constituted the AI group. After progression, 4 out of 5 patients remained receptor positive, while 1 patient lost ER expression. For PR, 3 out of 5 patients were initially PR positive. After AI, only 2 patients showed a positive PR status. 2 patients were PR negative before and after AI therapy. Conclusions: Contrary to the belief that clinical progression under ET is probably as a result of the loss of HR expression, our data suggests that progression under ET does not necessarily result in the loss of HR expression. In fact, 55% of relapses under Tam and 80% of the progressions under AI remain ER positive, indicating potential susceptibility to other endocrine therapeutic agents.

No significant financial relationships to disclose.
Relationship between thymidylate synthase expression and p53 levels with the treatment of cyclophosphamide, methotrexate, 5-fluorouracil chemotherapy(CMF) versus docetaxel (TXT) in locally advanced carcinoma of the breast
Authors: Brandi M. , Calascibetta A. , Cabibi D. , Martorana A. , Aragona F. , Fulfaro F. , Valerio M. , Dispenza J. , Sanguedolce R.
Published: March 2020

10546

Background: Adjuvant chemotherapy is used in the treatment of breast carcinoma independently of axillar node involvement. Different drug combinations such as CMF, FAC, FEC are still used; recently new drugs such as TXT (NEJM 332:1004,1997) show activity and are used also in adjuvant chemotherapy. 5 Fluorouracil (5Fu), a drug involved in main therapeutic regimens, blocks Thymidylate Synthase (TS), an enzyme involved in the DNA synthesis. TS not only links its own mRNA, but also p53 mRNA, inhibiting post transcriptional p53 protein synthesis. TS protein overexpression (Cancer Res 55:1407,1995), and/or its absence (Cancer Res 61:1421,2001) are some of the main mechanisms of 5-Fu drug resistance; moreover TS protein could inhibit p53 protein synthesis. Many clinical studies demonstrate that tumours with mutations of p53 are resistant to anthracyclines chemotherapy, 5Fu and radiotherapy (Carcinogenesis 17:1417,1996). Other studies show that the relationship between p53 and taxanes chemosensitivity seems to be more complex, because p53 mutations are associated with better taxanes drug response (Nature Med 2:255,1996). Aim of the study is to find association between TS and p53 levels, and relation with CMF or TXT treatment. Methods: We detected TS and p53 protein levels with immunohistochemistry assay in 84 paraffin embedded tumour samples from untreated patients (pts) who underwent surgery for primary breast cancer. Stage was T2N2M0 and pts were divided into two groups depending on the drug treatment received. One group (44 pts) was treated by 6 cycles of adjuvant CMF chemotherapy, the other by 4 cycles TXT alone. Results: 30% of pts showed lower TS and higher p53 levels, and had better TXT, while worse CMF treatment response; 30% of pts showed higher TS and lower p53 levels and all had worse response to all treatments; 40% of pts showed intermediate TS and p53 levels and different response to both CMF or TXT. Conclusions: On the basis of these results we identified a sub-group of pts showing lower TS and higher p53 levels that could be treated by a first line therapeutic regimen including Taxanes, not by drug combinations including 5 FU.

No significant financial relationships to disclose.
Adding paclitaxel to trastuzumab enhances antibody-dependent cell-mediated cytotoxicity via increment of natural killer cells in patients with Her-2 overexpressing breast cancer
Authors: Miura D. , Kitagawa D. , Tsutsumi K.
Published: March 2020

10548

Background: One of the mechanisms by which trastuzumab (TZ) inhibits the growth of Her-2 overexpressing breast cancer cells is the activation of a host tumor response via antibody-dependent cell-mediated cytotoxicity (ADCC). We questioned whether adding PTX to TZ enhances ADCC and also investigated kinetics of effector cells in ADCC. Methods: In ADCC, 5 cases with metastatic breast cancer receiving TZ (4mg/kg as a loading and 2 mg/kg weekly) with PTX (2qw, 100 mg/m2) were investigated. ADCC was analyzed by 51Cr releasing assay using SK-BR-3 cell line and white blood cells taken at the time of pretreatment, after 4 mg/kg of TZ, before/after 2 mg/kg of TZ, and before/after 2 mg/kg of TZ plus PTX. In of effector cell studies, we investigated fractions of natural killer (NK) cell, monocyte, and neutrophil taken at pre-administration and 10 minutes post-administration in 16 patients with Her-2 overexpressing breast cancer receiving weekly TZ and PTX (80 mg/m2). We defined NK cells as being both CD16+ (FcγRIII) and CD56+ and neutrophils as CD64+ (FcγRI) by flow cytometry. Results: Compared with pretreatment ADCC level, cytotoxicity were enhanced to 220% (median, ranging 30–259) after 4 mg/kg of TZ, 129% (78–210) after 2 mg/kg of TZ, and 148% (42–557) after the combination of TZ and PTX, whereas suppressed to 94% (48–163) after PTX alone. 2 weeks after the combination therapy, ADCC was significantly enhanced to 169% (113–257, p<0.05), compared with pre-treatment. In effector cells, NK cells increased in 131% (74–175, p<0.05) by TZ and in 224% (169–286, p<0.05) by the combination therapy. No significant changes were found in monocytes (98%, 50–160) and in neutrophils (100%, 50–160) by TZ alone, however monocytes decreased to 44% (34–50, p<0.05) and also neutrophils did to 49% (9–133) by the combination. Conclusions: Higher doses of TZ induced NK cell recruitment and the combination of TZ and PTX has a significant increase in recruitment and activation of NK cells. Adding PTX to TZ significantly enhanced ADCC via the rapid kinetics of NK cells. This may reflect that the combination of TZ and PTX has a strong synergistic effect than we expected in patients with Her-2 overexpressing breast cancer.

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The experience of a sample of Canadian men with breast cancer
Authors: Pituskin E. N. , Williams B. , Martin-Mcdonald K. , Au H.
Published: March 2020

10549

Background: Breast cancer is the most frequently diagnosed malignancy among North American women, accounting for about 30% of all new cases each year. Although male breast cancer accounts for less than 1% of all breast cancer cases, recent reports indicate significantly increasing incidence. Research suggests that men diagnosed with breast cancer experience serious psychologic issues including feeling less masculine by having a woman’s disease, feeling isolated with a rare malignancy and feeling uncomfortable seeking or receiving support. Men may also experience post-surgical effects including changed appearance of the male chest and decreased arm strength. There have been numerous large-scale studies examining womens’ experiences with breast cancer. To date, there have been no North American studies examining what a man experiences with a breast cancer diagnosis. The objective of this qualitative study was to determine the experiences of a sample of Canadian men diagnosed with breast cancer. Methods: After ethics approval, an invitation letter and consent form were mailed to those individuals listed as “male”, “breast cancer”, and “alive”, in the Alberta Cancer Registry. 20 men responded indicating their interest. After written consent, audiotaped interviews were conducted. As little is known about a man’s experience with breast cancer, an exploratory qualitative methodology was utilized. Results: We found a great variability among the participants with respect to whom they disclosed their diagnosis, how they viewed their masculinity after a breast cancer diagnosis and their views on life after a cancer experience. All participants stated that there was a major lack of awareness about male breast cancer, both in the public and health professional domain. All participants identified a lack of written information specific for men with breast cancer. All participants denied interest in traditional “support” groups, but would have welcomed the opportunity to speak with a man who had gone through similar diagnosis and treatment. Conclusions: Needs identified by the respondents include increased medical and public awareness of male breast cancer as well as specific written information. Further research is necessary to identify supports and resources helpful for male patients.

No significant financial relationships to disclose.
15-Hydroxyprostaglandin dehydrogenase (PGDH) is an aberrant tumor suppressor in human breast cancer
Authors: Wolf I. , O’kelly J. , Rubinek T. , Tong M. , Lin B. T. , Tai H. , Karlan B. Y. , Koeffler H. P.
Published: March 2020

10550

Background: Prostaglandin E2 plays a growth stimulatory role in breast cancer; and the rate-limiting enzyme in its synthesis, cyclooxygenase-2 (COX-2), is over-expressed in breast cancers. However, little is known about the expression pattern and activities of the key prostaglandin catabolic enzyme PGDH, in breast cancer pathogenesis. Methods: 5-aza-2’-deoxycytidine and suberoyl anilide bishydroxamide (SAHA) were used to unmask epigenetically silenced genes. DNA and RNA were extracted from tumors and cell lines using standard methods, and bisulfite sequencing was used for methylation analysis. Western blotting and real time PCR were used for expression analysis, and PGDH expression vector and PGDH-directed siRNA were used for overexpression and downregulation of PGDH, respectively. Athymic mice were utilized for xenograft models. Results: Using a pharmacologically based, genome-wide screen for epigenetically silenced genes, we found low levels of PGDH in MDA-MB-231 cells (estrogen receptor- (ER) and HER2-negative), but high levels in MCF-7 cells (ER-positive, HER2-negative), and observed upregulation of this enzyme following demethylation treatment. Methylation analysis revealed methylation of the PGDH promoter in one breast cancer cell line and in 3 of 10 primary tumors. Analysis of PGDH expression demonstrated low levels in 40% of primary breast tumors and identified a correlation between PGDH and ER expression. Upregulation of PGDH levels in MDA-MB-231 cells decreased their clonal growth and reduced their ability to form tumors in athymic mice. In contrast, silencing of PGDH in MCF-7 cells increased proliferation and enhanced in-vivo tumorigenicity. Further analysis revealed upregulation of aromatase following silencing of PGDH and inhibition of the ER pathway by forced expression of the enzyme. PGDH levels were downregulated by estrogen but upregulated by the tumor suppressor gene C/EBPα. Conclusions: Our results identify for the first time, that PGDH, the COX-2 antagonist, is aberrantly silenced in breast cancer. Activation of this enzyme, particularly in ER- and HER2-negative tumors, now becomes an interesting target for clinical studies.

No significant financial relationships to disclose.