bluuebunny/crossref_metadata_2025_split 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]