bluuebunny/crossref_metadata_2025_split huggingface dataset:
Differential protein kinase activity in ER-positive and ER-negative breast cancer
Authors: Ruijtenbeek R. , Umar A. , van Houten L. , Hilhorst R. , Foekens J. A. , de Wijn R. , Martens J. W. 
Published: August 2020

e22142 Background: Specific protein tyrosine kinases (PTKs) promote cancer progression and are potential drug targets. The presence of the estrogen receptor (ER) is an important criterion in deciding on treatment of patients with breast cancer. Since PTKs can affect the tumors' receptor function, we investigated in our first proof-of-principle study whether it is possible to discriminate ER-positive (ER+) from ER-negative (ER-) tumors based on PTK activity using microarrays containing many PTK peptide substrates. Methods: Cryosections of 12 ER+ and 12 ER- breast tumors were lysed in the presence of appropriate protease and phosphatase inhibitors. The protein kinase activity in the lysates was monitored in vitro using PamChip peptide microarrays, which comprise of 253 PTK peptide substrates derived from known human phosphorylation sites. Peptide phosphorylation through active kinases can be monitored in samples in real time, using a fluorescently labeled phospho-tyrosine specific antibody. Results: Phosphorylation activity profiles were determined in quadruplicate using multiple independent sample preparations. Using ANOVA analysis, out of 253 peptides, several peptides with a different phosphorylation signal in ER+ and ER- samples were found: 22 with p < 0.02 including VEGFR1, 2 and 3 and ADAM9. Multivariate unsupervised analysis using Principle Component Analysis showed a clustering of ER+ samples vs. ER- samples. Predictive analysis was performed using Partial-Least Squares Discriminant Analysis without explicit feature selection. The prediction error obtained from within experiment cross validation was typically in the range 10–20%. The error rate for between experiment prediction was 20%. An average predictive profile was constructed in which peptides with a relatively large weight were included, e.g. ADAM9 and BCAR1. Conclusions: Using PamChip peptide microarrays we have shown that differences in protein kinase activity exist between ER+ and ER- breast tumors. Our in vitro assay is a promising tool to investigate the interplay between kinase and nuclear receptor mediated signaling, with potential relevance to patient selection for targeted therapies. No significant financial relationships to disclose.
Comparative proteomic analysis of radiation induced changes in non small cell lung cancer patients with or without radiation induced lung toxicity
Authors: Cai X. , Shedden K. , Ao X. , Lawrence T. S. , Lubman D. M. , Kong F. 
Published: August 2020

e22146 Background: Radiation-induced lung toxicity (RILT) is dose limiting in the treatment of non small cell lung cancer (NSCLC). Early prediction of RILT would allow physicians to individualize treatment. This work aims to study if radiation induces differential changes in plasma proteomics in patients with and without RILT. Methods: 20 Patients with NSCLC in 3 stage- matched groups and treated with radiation therapy (RT) or chemoradiation were included in this analysis: 6 grade 0 (group 0), 8 grade 1 (group 1), and 6 grade ≥ 2 RILT (group 2). Platelet poor plasma was obtained pre-RT, at 2-, 4-, 6-week during-RT, and 1-, 3-month post-RT. The plasma proteomic characterizations from patients with and without RILT were compared using a multiplexed quantitative proteomics approach involving ExacTag labeling, reverse-phase high-performance liquid chromatography (RP-HPLC), and nano-LC electrospray tandem mass spectrometry (LC-ESI-MS/MS). ANOVA model was applied for significance test. Results: 23 common proteins were detected in all 20 patients, 9 of them, Ceruloplasmin, Prothrombin, Complement C3, Beta-2-glycoprotein (β2GP1) 1, Vitronectin, Complement factor I, Complement factor H, Complement C4-A, Complement C7, had significant difference among 3 groups. At baseline (Pre-RT), the levels of Vitronectin and C4-A in group 2 were both significantly higher than that in group 0 (p=0.008 and 0.021, respectively). At 2-week during RT, the levels of Ceruloplasmin, Prothrombin and β2GP1 in group 1 were significantly higher than that in group 0 (p=0.017, 0.011 and 0.013, respectively). At 4-week during RT, the levels of β2GP1 in group 1 and group 2 were significantly higher than that in group 0 (p=0.005 and 0.014). At 6-weeks during RT, Prothrombin, C3, Complement factor H, C7 and Complement factor I also had some significant changes among three groups. Conclusions: Plasma proteins related to inflammation, coagulation and fibrosis, such as β2GP1, change differently during RT in patients with and without RILT. This proteomic approach may identify new markers during treatment to predict RILT for adapting treatment to patient. Fundings: ASCO career developmental award, grant from Pardee foundation, and RTOG translational research program. No significant financial relationships to disclose.
Enhancer of zeste homologue 2 ( EZH-2) expression in breast cancer: a novel marker and potential target
Authors: Bandyopadhyay S. , Nahleh Z. , Ali Fehmi R. , Arabi H. , Sakr W. , Munkarah A. , Kruger M. 
Published: August 2020

e22154 Background: EZH-2 is a protein involved in cell cycle regulation; it belongs to the Polycomb group of proteins and has been suggested to be associated with aggressive breast cancer. In hormone receptor negative (HR-) breast carcinomas, novel therapeutic targets are needed. Few markers have achieved the success of Her-2/neu as targeted therapy making the search for novel treatment strategies imperative. In our study we investigate the expression of EZH2 in a cohort of hormone receptor negative breast carcinomas. Methods: We identified a consecutive cohort of 84 cases of HR- breast carcinoma in 2005–2006, from the Pathology department archives. Tumor grade, size, presence or absence of DCIS, lymph node status and Her2/neu expression were documented. The race of the patients was also noted. Immunohistochemical staining for EZH2 was performed on paraffin embedded sections. Nuclear expression of EZH2 was considered as positive and the percentage of cells staining positive was estimated. Using the Mann-Whitney U test, the expression of EZH2 was correlated with the tumor characteristics listed above. Results: The mean expression of EZH2 in HR- tumors was estimated at 74% with a median of 80% (0–90). Most of these cases (n= 61) were triple negative; 23 cases were HER-2/neu positive by established criteria. Increasing expression of EZH2 was correlated with increase in tumor size (>2 cm) and increased incidence of lymph node metastasis. No correlation was seen with tumor grade, the presence or absence of DCIS and Her2/neu expression. Race did not appear to have an impact on EZH2 expression. Conclusions: Our findings suggest that EZH2 is expressed in the majority of HR - breast cancer and is associated with aggressive breast carcinomas. EZH2 could be an important therapeutic target in this patient population. The study is ongoing to further characterize EZH2‘s role and its association with survival. No significant financial relationships to disclose.
Prognostic role of insulin-like growth factor receptor-1 (IGFR-1) and insulin-like growth factor binding protein-3 (IGFBP-3) expression in small cell lung cancer
Authors: Chang M. , Won Y. , Han J. , Kim H. , Kwon O. , Lee J. , Park Y. , Ahn J. , Ahn M. , Park K. 
Published: August 2020

e22155 Background: Insulin-like growth factor receptor-1 (IGFR-1) is a cellular membrane receptor overexpressed in many tumor cell lines and in some human tumors that seems to play a critical role in anti-apoptosis by enhancing cell survival. Also, insulin-like growth factor binding protein-3 (IGFBP-3) was reported to be a growth suppressor in variable pathways. Purpose of this study was to evaluate the state IGFR-1 and IGFBP-3 expression in patients with small cell lung cancer (SCLC) and its prognostic value. Methods: We analyzed IGFR-1 and IGFBP-3 expression in 194 SCLC tissues specimens by immunohistochemical stain. The relationship between IGFR-1 and IGFBP-3 expression and cliniopathological factors was evaluated. Univariate and multivariate analyses were performed to define its prognostic significance. Results: Median age was 63 years (range 38–85), 84% were men. One hundred-seventeen patients had extensive disease (60.3%), and 77 had limited disease (39.7%). With the median follow- up duration of 49.5 months (24–82), the median progression free survival (PFS) and overall survival (OS) were 8 months (95% CI: 7.3–8.7 months), and 14.4 months (95% CI: 12.7–16 months), respectively, The IGFR-1 expression was observed in 154 of 190 tumor tissues (79.4%), whereas there was no tissue stained by IGFBP-3. Multivariate analysis showed that stage (p<0.001), response to treatment (p<0.001), LDH level (p<0.001) were the independent prognostic factors for PFS, and age (p=0.014), LDH level (p<0.001), and stage (p<0.001) for OS. The IGFR-1 positivity was not associated with PFS or OS in the whole cohort. However, 84% of 115 extensive disease patients showed IGFR-1 positivity. The subgroup analysis revealed that OS was significantly longer for patients with IGFR-1 positive compared to those with IGFR-1 negative in extensive disease (11.3% vs 0% at 2year, p=0.034). Conclusions: These results suggest that IGFR-1 expression may be useful as a prognostic marker in patients with extensive disease of SCLC. No significant financial relationships to disclose.
Prostacyclin synthase (PTGIS) expression and epigenetic regulation in non-small cell lung cancer (NSCLC)
Authors: Gray S. G. , Cathcart M. C. , Al-Sarraf N. , Pidgeon G. P. , O'Byrne K. J. 
Published: August 2020

e22160 Background: Prostacyclin synthase (PTGIS) inhibits platelet aggregation and promotes vasodilation. Overexpression of this gene has been shown to inhibit lung cancer growth in a mouse model. Hypermethylation of the PTGIS promoter resulting in downregulation of PTGIS expression has been implicated in colorectal cancer. In this study we have examined both the expression patterns and epigenetic regulation of PTGIS in NSCLC. Methods: DNA/RNA and protein was extracted from matched tumour/normal samples (Thoracic Oncology Research Group BioBank, St James Hospital). PTGIS in these samples and a panel of retrospective resected lung samples was examined by immunohistochemistry and western blotting. A panel of NSCLC cell lines were treated with inhibitors to histone deacetylases (HDi) and DNA methyltransferases (DNMTi) and PTGIS expression examined by RT-PCR. Chromatin immunoprecipitation (ChIP) was used to examine changes to the PTGIS promoter in response to HDi. Results: PTGIS was found to have variable expression in both tumour samples and a panel of lung cancer cell lines. PTGIS expression was demonstrated in the vascular endothelial and bronchial epithelial cells of normal and cancerous sections. A striking variation in intratumoural PTGIS expression was observed. Methylation analysis of cell lines demonstrated hypermethylation of the PTGIS promoter. Treating cells with epigenetic inhibitors resulted in significant upregulation of PTGIS expression. Direct chromatin remodelling of the PTGIS promoter was confirmed. Conclusions: PTGIS is epigenetically regulated. The discrepancy between PTGIS mRNA and protein levels in tumor samples indicates that post-transcriptional and post-translational regulation of PTGIS is central to expression and requires further elucidation. Increased PTGIS expression is a potential therapeutic strategy for tumour prevention. [Table: see text]
Development of a new micro-injection-system for adenoviral gene delivery (MAGD)
Authors: Bohlen G. , Raabe A. , Dahm-Daphi J. , Dikomey E. , Schild S. E. , Dunst J. , Rades D. 
Published: August 2020

e22167 Background: Safe and effective delivery of viral vectors is important for gene therapy. Local application of such vectors is more efficient than systemic application. However, currently available injection systems are not optimal because placement of the injection needle and the vector application itself are generally not reproducibile. We developed a new injection system providing controlled injection of adenoviral vectors in tumors (xeno-transplants) in nude mice (NMRI nu/nu). Methods: The new MAGD consists of two fixation devices mounted on a 40x40cm plexi-glass plate, four injection units, and one pump unit with 4 infusion-pumps (Bee hive, Bioanalytical, Chesire, UK). Each injection unit is linked to a pump with commercially available plastic tube. Because the injection arms can be moved in x-,y- and z-direction, virus injection can easily performed for irregular tumor volumes. The entire injection volume is equally distributed to the 4 injection pumps allowing precise and steady injection rates between 0.1μl/min and 100μl/min. Success was confirmed with MRI-scans (Magnetom symphony, Siemens, Erlangen, Germany) 10 min after the injection. The efficacy of the MAGD was tested in 4 human squamous cell carcinoma cell lines from oro-/hypopharynx (FaDu, UD- SCC2, UD-SCC6, and UD-SCC7a). Virus transfection was performed with an adenovirus (serotype 5) expressing enhanced green fluorescent protein (eGFP). Transfection rates were quantified with flow cytometry. Successfully transfected cells expressed eGFP resulting in green fluorescence. In-vitro cell lines (concentration: 10 particles per cell, MOI 10) of FaDu, UD-SCC2, UD-SCC6 and UD-SCC7a served as controls. Results: The MAGD was easy to handle. Injection of 100μl (25μl per infusion pump) at an injection rate of 5μl/min took only 5 min. The in-vivo transfection rates achieved with the MAGD were 9±1% for FaDu, 39±2% for UD- SCC, 54±1% for UD-SCC6 and 54±9% for UD-SCC7a, respectively. The in-vitro transfection rates (controls) for these 4 cell lines were 2±1%, 23±4%, 29±4% and 3±2%, respectively. Conclusions: The new MAGD provided controlled injection of adenoviral vectors in tumors in nude mice. It proved to be effective, as it resulted in comparably high transfection rates. No significant financial relationships to disclose.
Identifying the primary site using gene expression profiling in patients with carcinoma of an unknown primary (CUP): a feasibility study from the GEFCAPI
Authors: Loriot Y. , Gross-Goupil M. , Lesimple T. , Blot E. , Merrouche Y. , Mathieu M. C. , Fizazi K. 
Published: August 2020

e22168 Background: CUP are an heterogeneous family of neoplasms with a dismal prognosis, with empiric chemotherapy as the recommended treatment. The aim of this study was to evaluate the feasibility of a 500-mRNA microarray to identify the tissue of origin in patients with CUP. Methods: Diagnostic biopsy formalin-fixed, paraffin-embedded (FFPE) specimens from 22 patients with CUP were prospectively collected. Gene expression profiling was performed using oligonucleotide microarray that contains 495 genes selected as highly differentially expressed between 49 tumor types (CupPrint). Results: The assay was successfully performed on specimens from 18 of the 22 patients (82%). It could not be performed because of a low RNA preservation in the remaining 4 cases. The median age was 57 years (range: 29–70 years). The median delay from tissue shipping to receipt of CupPrint result was 11 days (range: 1–26 days). The most common tissues of origin identified were lung cancer (22%) and colorectal cancer (17%). Of note, a primary cancer which would not be adequately treated by an empiric chemotherapy regimen currently recommended in CUP (like cisplatin-gemcitabine or carboplatin-paclitaxel) was identified in about half patients: kidney cancer (1), hepatocarcinoma (1), colorectal cancer (3), head and neck cancer (2) and cholangiocarcinoma (1). Conclusions: Gene expression profiling of FFPE biopsy specimens from patients with CUP is feasible in a reasonable delay, making it feasible in clinical practice. A phase III randomized trial is planned to compare therapy based on gene expression-suspected primary cancer versus empiric chemotherapy. No significant financial relationships to disclose.
Predictive and prognostic functions of microtubule-associated protein-tau and topoisomerase IIα protein in early breast cancer
Authors: Choi M. , Won H. , Lee K. , Sung S. , Moon B. , Choi H. , Suh H. , Nam E. , Lee S. 
Published: August 2020

e22169 Background: Topoisomerase IIα protein(topo II) is the molecular target of topo II inhibitors such as anthracyclines and Microtubule-Associated Protein(MAP)-tau protein is associated with taxane sensitivity. Anthracyclins and taxanes are major cytotoxic agents of breast cancer in the adjuvant setting. The aim of this study was to evaluate the predictive and prognostic functions of MAP-tau and topo II in early breast cancers. Methods: Representative breast tumor sections were constructed from paraffin embedded specimens from 78 node positive breast cancer patients. MAP-tau and topo II protein were assessed by immunochemistry using antibody clone 4F1(Affinity BioReagents,USA) and clone Ki-S1 antibody(Dakocytomation,USA). MAP-tau staining of tumor cells was semiquantatively scored as 0, 1+, 2+, 3+ and cases with 0 or 1+ staining intensity were considered MAP-tau negative. Topo II protein over- expression was defined as the detection of nuclear staining in more than median value of evaluated cells. Results: Thirty- four cases (43.6%) of 78 samples showed topo II overexpression and 35 cases(44.9%) showed MAP-tau overexpression in node positive breast cancers. HER2 overexpression was noted in 28 samples (35.9%) and 56 cases (71.8%) were compatible with the luminal type. In 43 patients (55.1%), anthracyclin and taxane were used as adjuvant therapy and in this group, both MAP-tau and topo II overexpression showed lower disease-free survival (DFS) than the others, but statistically not significant. In luminal type, MAP-tau overexpression was poor prognostic factor on DFS in Cox regression.(HR 5.644, 95% CI 1.14–28.07, p=0.034) Conclusions: Topo II overexpression and MAP-tau overexpression in node positive breast cancers were not significant predictive factors for anthracyclin and taxane therapies. As several investigators reported, MAP-tau is associated endocrine therapy sensitivity in patients without chemotherapy, but higher MAP-tau in luminal type was a strong poor prognostic factor in patients who were given chemotherapy and hormonal therapy. No significant financial relationships to disclose.
TP53 codon 72 polymorphisms and breast carcinoma susceptibility: A meta-analysis
Authors: Zhu B. , Zhuo W. , Chen Z. 
Published: August 2020

e22171 Background: Previously, TP53 codon 72 polymorphisms have been implicated as risk factors for various cancers. Several studies have conducted on the association of TP53 codon 72 polymorphisms with susceptibility to breast carcinoma and have yielded inconclusive results. The aim of the present study was to assess possible associations of breast cancer risk with TP53 codon 72 polymorphisms. Methods: We conducted a search in the Medline, EMBASE, OVID, Sciencedirect, and Chinese National Knowledge Infrastructure (CNKI) without a language limitation, covering all papers published up to Dec 2008. The associated literature was acquired through deliberate searching and selected based on the established inclusion criteria for publications. Results: Consequently, fifteen studies, including 3436 cases and 4394 controls, met the included criteria and thus were selected. Ultimately, the relevant data were extracted and further analyzed using systematic meta- analyses. The results showed that individuals carrying homozygote Arg/Arg genotype have a significant increased risk of breast cancer compared with those carrying Pro/Pro genotype (OR: 1.58, 95%CI:1.10–2.28). For Arg allele, no evidence indicated that individuals with Arg/Arg genotype have an increased risk of breast cancer compared with those with a combined Pro genotype (Arg/Pro+Pro/Pro) (OR: 1.68, 95%CI:1.24–2.29). For Pro allele, individuals with homozygote Pro/Pro genotype have a marked decreased susceptibility to breast cancer relative to those with a combined Arg genotype (Arg/Pro+Arg/Arg) (OR: 0.84, 95%CI:0.73–0.98). Conclusions: The results of the present study suggest that TP53 codon 72 polymorphisms might be a risk factor for breast cancer. Homozygote Arg allele genotype could significantly increase susceptibility to breast cancer, while Pro/Pro allele markedly decreases breast risk. No significant financial relationships to disclose.
Malignant gastrointestinal stromal tumors treated with imatinib in France: Results in unselected patients
Authors: Mathieu-Boue A. , Bouché O. , Cioffi A. , Rios M. , Duffaud F. , Chaigneau L. , Ray-Coquard I. , Blay J. , Le Cesne A. , Deschaseaux C. 
Published: August 2020

e22177 Background: GISTs are rare tumors of the GI tract. In France, their incidence is estimated to be 9- 12/106 inhabitants/year. Imatinib (IM) has been approved to treat unresectable and/or metastatic Kit-positive GISTs since 2002 but information on routine use, safety and efficacy in unselected “real life” setting is lacking. An observational cohort (EPIGIST) in France was designed to provide data on survival, safety and treatment patterns and quality of life. Methods: EPIGIST is a nationwide multi-center, observational study on GIST patients (pts) treated with IM for the first time between the availability on the French Market and the end of the 2008. Centers were randomly selected in national files of oncologists, gastrointestinal surgeons and gastrointestinal specialists. The planned follow-up duration was three years. A case report form (CRF) had to be completed at inclusion and during each follow-up visits. Quality of life was assessed using QLQ-C30 and SF36 questionnaires. Results: 29 on 51 selected centers enrolled at least one pt and 127 pts were included (as of 12/2008), The median age of disease onset was 59 years (range 29 - 85) with 48% pts>50 years. 63% were symptomatic at diagnosis; 69% were fully active (grade 0 on the ECOG index). Primary tumors were most often gastric (34%), or from jejunum/ileum (24%). At diagnosis 84% of pts had a tumor size over 5cm. 64% of patients had surgery of the primary tumor before starting IM. For 86% of the pts, IM was given at an in initial dosage of 400 mg, 8% at 300 mg and 6% at 800mg. Compliance was superior to 90% for all pts. With a median follow-up of 1.72 years (CI95%: [1.08;1.95]), two-years overall survival from first treatment with IM was 88.1% (CI95%: [65.3%-96.3%]). Conclusions: EPIGIST is still an ongoing survey. Current results confirm previous published data on survival in GIST treated with IM in an unselected cohort of patients outside of a clinical trial. [Table: see text]