bluuebunny/crossref_metadata_2025_split huggingface dataset:
Concordance of local and central laboratory hormone and HER2 receptor status in ECOG 2197
Authors: Badve S. S. , Baehner F. L. , Gray R. , Childs B. , Maddala T. , Rowley S. , Shak S. , Davidson N. , Goldstein L. J. , Sparano J. 
Published: March 2020

21022 Background: Central and local laboratory concordance for hormone and HER2 receptor measurement is of national interest. This study compares ER/PR/HER2 by local laboratories using immunohistochemistry (IHC) and central laboratories (IHC & quantitative RT-PCR). Methods: Of 2952 patients in E2197, a case-cohort sample of 776 patients who either did (N=179) or did not recur was studied. Central IHC for ER/PR/HER2 was performed using single 0.6 mm microarrays; Allred score (AS) was used for ER/PR (AS>2 = positive). Positive HER2 was 3+ staining in >10% cells for Central IHC and 2+ or 3+ for Local IHC. RT-PCR analysis by Oncotype DX™ for ER/PR/HER2 was performed using pre-defined cutoffs of 6.5, 5.5 and 11.5 units, respectively. Hormone receptor (HR) pos was defined as ER &/or PR pos. Results: Results from Local IHC (ER/PR in 776 & HER2 in 517 pts) were compared with Central IHC (760 pts) and RT-PCR results (776 pts). The discordance between HR positivity by Local IHC and RT-PCR was very low. However, 12% of HR neg pts by Local IHC (38/321) & Central IHC (39/326) were HR pos by RT-PCR. The relationship between ER and recurrence as a function of AS was examined. Patients with AS of 3–4 were found to be closer to the AS=2 group than to the AS>4 group Patients with AS of 3–4 were found to be closer to the AS ÿ 2 group than to the AS > 4 group (Est.HR for ER 0.97 for AS 3–4 vs. 0–2 and 0.46 for AS 5–8 vs. 0–2, and for PR were 0.84 for AS 3–4 vs. 0–2 and 0.41 for AS 5–8 vs. 0–2). Conclusions: There is a high degree of overall concordance among Local IHC, Central IHC, and Central RT-PCR for ER and PR. The degree of concordance is even greater for HR compared to ER or PR alone. Although the concordance with local labs for HER2 testing was poor, the concordance between Central IHC and RT-PCR was very high. The relatively high incidence (12%) of IHC HR neg pts who are HR pos by RT- PCR is notable. [Table: see text] [Table: see text]
Detection of DNA methylation in inflammatory breast tumours by a multiplex nested methylation-specific PCR
Authors: Van der Auwera I. , Van den Bosch S. , Van Laere S. , Van den Eynden G. , van Dam P. , Colpaert C. , van Engeland M. , Van Marck E. , Vermeulen P. , Dirix L. 
Published: March 2020

21027 Background: Hypermethylation of gene promoters is a common mechanism of loss of gene function in cancer cells. Few data on epigenetic alterations in inflammatory breast cancer (IBC) cells are available. Microarray studies have shown that IBC is characterised by a distinct gene expression profile when compared to non-IBC. We sought to determine whether the profiles of gene hypermethylation differ between IBC and non-IBC by using a multiplex nested methylation-specific PCR (MSP). We selected genes for which evidence exists for their involvement in breast carcinogenesis by means of methylation. Methods: The SLμCut system (MMI) was used for laser capture microdissection of paraffin-embedded tissues. 10-μm-thick sections from 10 patients with IBC and 12 patients with non-IBC were used. Genomic DNA from the microdissected tissue was extracted and treated with sodium bisulfite. The methylation status of the promoter regions of APC, E- cadherin, THBSP-1, DAPK, GSTP1, RASSF1A and BRCA1 were investigated using a multiplex nested MSP method. DNA isolated from normal peripheral lymphocytes from healthy individuals served as a negative methylation control. In vitro methylated DNA served as the positive methylation control. MSP products were analyzed on agarose gel electrophoresis, and were determined to have methylation if a visible band was observed in the methylation reaction. Results: Methylation of the APC gene promoter was observed in 33.3% (4/12) of non-IBC samples and 70% (7/10) of IBC samples. Methylation of the BRCA1 gene promoter was observed in 16.7% (2/12) of non-IBC samples and 30% (3/10) of IBC samples. Methylation of the DAPK gene promoter was observed in 16.7% (2/12) of non-IBC samples and 10% (1/10) of IBC samples. Methylation of the GSTP1 gene promoter was observed in 33.3% (4/12) of non-IBC samples and 30% (3/10) of IBC samples. No methylation of the E-cadherin and THBSP-1 gene promoters was observed in non-IBC samples or IBC samples. Conclusions: Also in this study, the multiplex nested MSP approach was proven to be very suitable for the detection of methylation in samples harbouring small amounts of poor quality DNA. In the near future, the methylation status of other genes will be determined and the study population will be enlarged. No significant financial relationships to disclose.
EBV-positive lymphoma patients have a selective deficiency in EBV immunity
Authors: Heller K. N. , Steinherz P. G. , Portlock C. S. , Münz C. 
Published: March 2020

21032 Background: Epstein-Barr virus (EBV) asymptomatically establishes persistent infections in more than 90% of the adult population. However, due to effective immune control, only a minority of infected carriers develops spontaneous EBV-associated lymphomas. Since EBV nuclear antigen-1 (EBNA1) is the only protein expressed in all proliferating EBV infected cells we hypothesize that EBNA1 specific immune response is critical in preventing EBV-positive lymphomas. Methods: After informed consent, peripheral blood from healthy volunteers and lymphoma patients (prior to therapy- no evidence of cytopenia) were stimulated (ex vivo) with overlapping peptides covering the immunogenic EBNA1 (aa400–641) sequence. Frequency of EBNA1-specific T-cells were assessed by intracellular cytokine staining and flow cytometric proliferation assays. Cytokine pattern, surface marker phenotype and functional reactivity against EBV specific and control antigens were analyzed. Results: Patient and volunteer immune responses to control antigens and other viruses were assessed and statistically indistinguishable. EBNA1 specific CD4+ T cell responses were detected among 18 of 20 healthy carriers, and among 10 of 16 patients with EBV-negative lymphoma (relative to healthy volunteers p=0.145 via paired student T test). None of the patients with EBV-positive lymphomas (n=8) had a detectable EBNA1-specific CD4+ T-cell response (p<0.003 relative to healthy volunteers and patients with EBV-negative lymphomas). Conclusions: Healthy volunteers and patients with EBV-negative lymphoma have statistically similar EBNA1-specific CD4+ T cell responses. Although patients with EBV-positive lymphoma have intact immune responses to common viruses and antigens, they selectively lack an EBNA1-specific CD4+ T cell response. An intact EBNA1 specific immune response among patients with EBV-negaitve lymphoma implies that lymphoma is not a cause of a selective immune deficiency. On the contrary, these findings suggest that EBNA1-specific CD4+ T cells are critical in the prevention of EBV mediated lymphomas, and a defect in EBNA1 specific immunity may leave EBV carriers suseptible to EBV-positive lymphomas. EBNA1- specific CD4+ T cell function may be a new target for therapies of EBV-associated malignancies. No significant financial relationships to disclose.
Proliferation and apoptosis before and after primary systemic therapy for operable breast cancer
Authors: Guarneri V. , Piacentini F. , Frassoldati A. , Ficarra G. , Jovic G. , Puglisi F. , Michelotti A. , Bisagni G. , Berardi R. , Conte P. 
Published: March 2020

21045 Background: The identification of biological markers able to predict tumor sensitivity to primary systemic therapy (PST) may have an important clinical output. Moreover, the effect of treatment on these biomarkers might have prognostic relevance. Aims of this analysis: to evaluate the predictive role of proliferation (Ki-67), apoptotic index (AI) and growth index (GI) before PST, the effect of PST on these parameters and their relation with tumor response and patients outcome. Methods: Ki-67, AI (Tunel Test), and GI were assessed at baseline and at surgery in 90 stage II-IIIB breast cancer patients enrolled in a phase II randomized trial of PST with 4 cycles of epirubicin-paclitaxel ± gefitinib. Pathologic response was defined as tumor regression grade (TRG) (criteria of Miller and Payne). Results: patients characteristics were as follows: median age, 51 yrs (range 29–69); stage IIA 41%, IIB-IIIA 59%. A significant correlation between Ki-67 and AI was found at baseline (p=0.001). A significantly higher Ki-67 and AI were observed in ER- vs ER+ tumors (median Ki-67: 62% vs 25%, p<0.0001; median AI: 1.33 vs 0.85, p=0.009). A significantly higher Ki-67 and GI were observed in EGFR+ vs EGFR- tumors (median Ki-67: 60% vs 25%, p=0.0006; median GI: 48 vs 30.7, p=0.03). No significant differences were observed according to HER2 expression. After PST, 84% of pts was classified as having a TRG 4 or 5, and 16% as having a TRG 1–3. No significant correlation among baseline Ki-67, AI and GI and TRG was observed. After PST, Ki 67 and AI were significantly reduced from baseline. Lower Ki 67 and lower AI were significantly correlated with higher TRG (p=0.008 and 0.01 respectively). We observed 1 cancer death and 5 relapses only, therefore correlation with patient outcome has been planned for the time of the meeting Conclusions: ER- and EGFR+ tumors show more aggressive behaviour. PST significantly reduces Ki-67 and AI, and this reduction is significantly correlated with TRG. In particular, the reduced AI at surgery was not found to be a marker of resistance, more probably reflecting the reduced Ki-67 of responding tumors. No significant financial relationships to disclose.
EGFR expression is associated with increased cell turnover and poor histologic grade in human colon carcinomas
Authors: Rego R. L. , Smyrk T. C. , Foster N. R. , Sargent D. J. , Kabat B. F. , French A. J. , Thibodeau S. N. , O'Connell M. J. , Windschitl H. , Sinicrope F. A. 
Published: March 2020

21046 Background: The Epidermal Growth Factor Receptor (EGFR) is a transmembrane receptor tyrosine kinase that is important for colon cancer cell growth and represents a therapeutic target. We studied EGFR expression in relation to rates of apoptosis and cell proliferation. Methods: TNM stage II and III colon carcinomas (n= 361) from patients treated in a 5-FU-based adjuvant trial were studied. EGFR expression (EGFR pharmDx™) and extent were analyzed and their relationship to rates of cell turnover, histologic grade, mismatch repair (MMR) status (defective if instability at BAT 26 locus plus loss of hMLH1, hMSH2 or hMSH6 proteins) and clinical outcome were determined. Results: EGFR plasma membrane staining was detected in 214 (59.3 %) colon carcinomas. Increased EGFR intensity (2+, 3+) was associated with a higher number of caspase-3- (p= 0.02) and Ki-67- (p< 0.001)positive tumor cells. Increased EGFR intensity (p= 0.02) and extent (p= 0.03) were associated with poor differentiation, and did not depend upon MMR status (Breslow-Day test p ≥ 0.46). Diffuse (vs. focal) EGFR extent was increased in tumors with defective MMR [22 of 32 (69%)] compared to those with intact MMR [136 of 271 (50%)] (p= 0.047). EGFR intensity was unrelated to MMR status. Neither EGFR intensity nor extent was significantly associated with patient survival. Conclusions: Stage II and III colon cancers with EGFR overexpression show adverse prognostic variables, i.e., increased rates of cell turnover and poor histologic grade, suggesting that EGFR may be an important target for adjuvant therapy. *Chi-square test for categorical data, Wilcoxin rank-sum test for continuous data [Table: see text] No significant financial relationships to disclose.
Role of MSI and DCC status to predict response to FOLFOX in metastatic colorectal cancer
Authors: Bennamoun M. , Schischmanoff O. , Martin A. , Mariani P. , Ah Koon L. , Morere J. , Des Guetz G. 
Published: March 2020

21056 Background: Microsatellite instability (MSI) and lack of Deleted Colon Cancer (DCC) protein expression have been observed respectively in ∼15% and 70% of colon cancer cases and are considered as prognostic factors in colorectal cancer (CRC). In adjuvant setting these markers could be considered to decide treatment. We know that MSI colon cancer do not benefit from 5Fluorouracyl (5FU)-based chemotherapy. A current treatment of reference for colon cancer is a combination of 5FU and Oxaliplatin (FOLFOX). Our aim was to determine whether MSI status or DCC expression are predictive markers of FOLFOX efficiency in patients with metastatic CRC. Methods: Tumour specimens were collected from patients with metastatic colon cancer treated with FOLFOX. The FOLFOX regimen was evaluated in first line treatment according to the WHO criteria. The microsatellite instability status was assessed by measurement of the length of five monomorphic mononucleotide markers. DCC protein expression were evaluated by immunohistochemistry. Results: Forty patients (22 men, 18 women), median age 63.5 years (27–83) were treated with FOLFOX. Nine patients had tumours exhibiting high microsatellite instability (microsatellite instability group, MSI group) and 31 patients had tumours exhibiting microsatellite stability (microsatellite stable group, MSS group). In MSS group, we observed 11 partial responses (36%) and only two in MSI group (22%) (not significant difference). Both disease free survival and overall survival were not significantly different for MSI and MSS groups. Determination of DCC status is under investigation. Conclusions: There was no significant difference in chemosensitivity to FOLFOX for MSI and MSS metastatic patients. Therefore FOLFOX regimen could be proposed to metastatic patients irrespective of MSI status. The study of the relationship between DCC expression and response to FOLFOX is ongoing. No significant financial relationships to disclose.
Differential gene expression profile in breast cancer-derived stromal fibroblasts
Authors: Singer C. F. , Gschwantler-Kaulich D. , Fink-Retter A. , Hudelist G. , Haas C. , Czerwenka K. , Kubista E. , Kubista E. 
Published: March 2020

21075 Background: Breast cancer is chatacterized by malignant transformation of epithelial cells, but stromal cells also play an important role in tumorigenesis. While tumoral fibroblasts display unique phenotypical properties, it is unclear whether they also represent are a specific subpopulation. Materials and Methods: Stromal fibroblasts deriving from malignant tissue of 10 women with invasive breast cancer, and from normal breast tissue of 10 women with benign breast disorders, were subjected to differential complementary DNA Microarray Analysis by using a 2400 gene cDNA array. Gene expression results were validated by real-time PCR and by immunohistochemistry. Results: In a cDNA Array that allows to analyze the differential gene expression of more than 2400 genes, the mRNA expression of 135 genes were increased more than 2 fold in fibroblasts from malignant breast tumors. The majority of these genes encode tumor-promoting cytokines, transcription factors and cell-matrix associated proteins. The mRNA expression of 110 genes decreased to less than 0.5 fold. The remaining 2155 genes were not significantly altered. Immunohistochemistry for selected proteins performed on biopsies from breast cancer and normal breast tissues confirmed the clinical relevance of our findings. Conclusion: Breast cancer-derived stromal fibroblasts show a distinctive gene expression pattern that differentiates them from normal breast stroma. Our observation of increased expression of tumor promotion-associated genes even in the absence of adjacent malignant epithelium suggests that tumor stroma is comprised of a fibroblastic subpopulation that provides for a microenvironment which supports tumor growth and invasion. No significant financial relationships to disclose.
Pattern of expression of calpain subunits (large and small) in human breast cancer and the prognostic significance
Authors: Chhabra A. , Fernando H. , Mansel R. E. , Jiang W. G. ,   Metastasis & Angiogenesis Research Group
Published: March 2020

21078 Background: Calpains belongs to family of non-lysosomal calcium dependent cysteine proteases which are known to regulate cellular migration, apoptosis and cell cycle progression in normal and tumour cells. Calpain has also been indicated in the metastatic process of certain tumours like RCC and identified as a potential tumor suppressor for gastric cancer. Little is known about the expression pattern of calpain in human breast cancer. The current study investigated the expression pattern of the large (calpainL) and small (calpainS) subunits of calpain which are encoded by different genes, in human breast cancer and attempted to correlate the expression with clinical outcome. Methods: RNA was extracted from frozen sections of breast tissue (n=120, median clinical follow up of the patients - 72 months) for gene amplification. The expression of both subunits of calpain was determined by using RT-PCR and quantitative RT-PCR. Statistical analysis was carried out using Mann- Whitney U test and the Kruskal- Wallis test. Results: We found significantly higher level of both calpain subunits in tumour tissue (n=120) as compared to normal non-neoplastic mammary tissues (calpainL, p=0.022 and calpainS, p=0.039). The expression of calpainL was significantly lower in patients with poor clinical outcome (with metastasis, p=0.024, with local recurrence, p=0.024 and who died of breast cancer, p=0.028), than those who were disease free. In contrast, the levels of calpainS were high in patients with poor prognosis, metastasis and who died of breast cancer but were statistically not significant. The expression of calpainL was marginally lower in node positive tumours than the node negative tumours. CalpainS transcripts were higher in node positive than node negative tumours (p=0.23). Similarly, calpainL showed a low level in high grade tumours whereas calpainS displayed a contrasting high level in these tumours. Conclusions: The large and small subunits of calpain have a distinct pattern of expression in human breast cancer. A decrease expression of calpainL but an increase expression of calpainS may be associated with poor prognostic tumours, thus indicating the intimate nature of the molecular pair in the disease progression of human breast cancer. No significant financial relationships to disclose.
Identification of a new subset of myeloid suppressor cells in peripheral blood of melanoma patients and modulation by GM-CSF-based anti-tumor vaccine
Authors: Filipazzi P. , Valenti R. , Huber V. , Iero M. , Pilla L. , Parmiani G. , Santinami M. , Rivoltini L. 
Published: March 2020

21082 Background: Phenotypic and functional features of myeloid suppressor cells (MSC), known to serve as critical regulators of anti-tumor T cell responses in tumor-bearing mice, are still poorly defined in human cancers. Here we analyzed myeloid subsets with suppressive activity present in peripheral blood of metastatic melanoma patients (MM) and evaluated their modulation by a GM-CSF-based anti- tumor vaccine. Methods: Stage IV AJCC MM patients (n=16) vaccinated with autologous tumor-derived heat-shock protein peptide complexes gp96 (HSPPC-96) and low dose GM-CSF provided pre- and post-treatment whole blood samples. Peripheral blood mononuclear cells (PBMC) were analyzed by flow cytometry, separated into cellular subsets and used for in vitro proliferation assays. PBMC from stage- matched melanoma patients (n=12) treated with non-GM-CSF-based vaccines (i.e. HSPPC-96 alone or IFNa/melanoma-derived peptides) or gender and age-matched healthy donors (n=16) were also analyzed for comparison. Results: The lack or low levels of HLA-DR expression was found to identify a CD14+ cell subset with high suppressive activity on lymphocyte proliferation and functions. CD14+HLA-DR-/lo cells were significantly expanded in all MM patients, while undetectable in healthy donors. Suppressive activity was mediated by TGFβ, as suggested by functional experiments with neutralizing specific antibodies. In contrast, no involvement of arginase and iNOS pathways could be detected. CD14+HLA-DR-/lo cells, as well as spontaneous ex- vivo release and plasma levels of TGFβ, were augmented after administration of the HSPPC-96/GM-CSF vaccine. Interestingly, the expansion of suppressive CD14+ monocytes was associated to the inability to mount a significant CD8-mediated T cell response upon vaccination. On the other hand, no quantitative or qualitative enhancement of the CD14+HLA-DR-/lo suppressive cell population was observed in patients receiving a non-GM-CSF based vaccine. Conclusions: CD14+HLA-DR- /lo cells exerting TGFβ-mediated immune suppression may represent a new subset of myeloid suppressive cells, potentially expandable by the administration of GM-CSF-based vaccines in melanoma patients. [Table: see text]
Overexpression of VEGF-C and -D may predict unfavorable outcome in resected non-small cell lung cancer
Authors: Ko Y. , Jung C. , Lee M. , Kang J. , Hong Y. , Lee K. 
Published: March 2020

21114 Background: Correlation of Vascular endothelial growth factor (VEGF)-C, VEGF-D and/or VEGF receptor 3 (VEGFR-3) with lymphatic spread, tissue invasion and poor clinical outcomes has been observed in various cancers. Nodal metastasis is known to one of poor prognostic factors in non-small cell lung cancer (NSCLC). We investigated the correlation of VEGF-C,-VEGF-D and VEGFR-3 with clinicopathologic parameters and patient survival in NSCLC. Methods: Using immunohistochemial staining, we analyzed the protein expressions of VEGF-C, VEGFD and VEGFR-3 on the tissue array specimens from 180 patients with completely resected NSCLC. A 0 to +1 immunohistochemical staining (IHCS) of the cancer cell was defined as negative, +2 to +3 IHCS was as positive. Results: The expressions of VEGF-C, VEGF-D and VEGFR-3 were observed in 28.9%, 32.8% and 30.6% of cases, respectively. In the Cox regression-based multivariate analysis, it was proved that VEGF-C and D proved to be an independent prognostic factors as well as known prognostic factors, such as tumor size and lymph node metastases (VEGF-C, p=0.001; VEGF-D, p=0.014). VEGF-C expression showed statistically significant correlation with the expression of VEGFR-3 (p=0.02). Conclusion: To be taken, the present study revealed that VEGF-C and D expression may predict poor prognosis in resected NSCLC patients. Therefore, these results seems to be worth developing promising targeting inhibitors on lymphangiogenesis which plays major role in locoregional recurrence for surgically NSCLC patinets. No significant financial relationships to disclose.