bluuebunny/crossref_metadata_2025_split huggingface dataset:
Use of microRNAs to distinguish small from non-small lung cancer
Authors: Spector Y. , Meiri E. , Faerman A. , Ben David M. , Zepeniuk M. , Ezagouri M. , Cohen D. , Rosenwald S. , Perelman M. , Barshack I. 
Published: August 2020

e22061 Background: Lung tumors are divided to two main classes: non-small cell lung cancer (NSCLC) that accounts for ∼80–85% of all lung primary tumors, and tumors from neuroendocrine origin - mainly small cell lung carcinoma and lung carcinoid. The classification of lung tumors can present a diagnostic challenge. New markers for different subtypes may aid in diagnosing difficult cases, can improve the accuracy of classification and could be important for selecting proper treatment. Here we studied the utility of microRNA as biomarkers for this differential diagnosis. MicroRNAs, a family of short non-coding regulatory RNAs, are highly tissue-specific and are well preserved in routinely prepared formalin-fixed, paraffin-embedded (FFPE) specimens, making them promising candidates as biomarkers for tissue and tumor classification. Methods: We used proprietary protocols for extracting high-quality RNA from FFPE samples. We used microRNA microarrays to profile more than a hundred samples from different histological subtypes of lung cancer including small cell, lung carcinoid and various types of NSCLC. Differential microRNA expression was verified using a microRNA qRT- PCR platform. Results: We found that several microRNAs are significantly differentially expressed between different subtypes of lung cancers. Specifically, using combinations of few microRNAs, we were able to accurately differentiate between neuroendocrine and NSCLC. Small cell and carcinoid tumors can be further distinguished using the signals of additional microRNAs, with very high sensitivity and specificity. Conclusions: Our results underscore the potential of microRNA expression for classification of tumor subtypes. We found that combinations of small numbers of microRNAs can successfully aid in the differential diagnosis of lung tumors, and provide a basis for the development of simple and reliable assays for clinical oncology. [Table: see text]
Development of ultra-sensitive single molecule immunoassays for the intracellular kinases Akt1/pAkt1 and Erk2/pErk2 for small tissue studies
Authors: Todd J. , Held D. , Klein B. K. , Macy E. , Agee S. J. , Lu Q. A. , Puskas R. 
Published: August 2020

e22068 Background: The Ras/Raf/MEK/ERK and PI3K/PTEN/AKT kinase cascades are critical pathways in malignant transformation, tumor progression and multi-drug resistance (MDR) in human cancers. Expression and genetic studies have contributed much to our understanding of this role, but further clinical investigations at the post-transcriptional level are necessary. Towards this end, Singulex has developed single molecule immunoassays for total Akt1, pAkt1, total Erk2 and pErk2 utilizing the Erenna Immunoassay System for use in small tissue studies and clinical trials. Methods: Analytical sensitivity, inter- assay precision, linearity and specificity were determined and compared to corresponding ELISA assays. Novel assays were tested on lysates from a variety of commonly utilized cell lines in cancer studies: MCF-7, PBMC, Jurkat, HeLa, and NIH3T3 cells. Results: All four novel assays were significantly more sensitive than the ELISA assay method based upon sample endpoint dilutions. Acceptable precision (<20%CV) was demonstrated for the Akt1/pAkt1 assays in PBMC lysates and for the Erk2/pErk2 assays in MCF-7, HeLa and NIH3T3 lysates. The LLoQ of the Akt1/pAkt1 assay in Jurkat lysates was sufficient to quantify Akt1/pAkt1 from as few as 10–20 cells. Based upon a typical 1mm needle biopsy, this is sufficient to quantify Akt1/pAkt1 from less than 1/1000th of that biopsy. Conclusions: We show that the Singulex Akt1, pAkt1, Erk2 and pErk2 assays are over 50 fold more sensitive than currently available ELISA assays, and specifically quantify analyte in a variety of cell lysates. We have further demonstrated that the Akt1/pAkt1 assay can quantify from as little as 1/1000th of a typical needle biopsy. Thus, these novel assays greatly improve upon pre- existing technology, providing a new tool for clinical trials investigating Ras/Raf/MEK/ERK and PI3K/PTEN/AKT pathways previously limited by ELISA sensitivity. [Table: see text]
Evaluation of serum profiles changes after neoadjuvant chemotherapy for breast cancer using MALDI-TOF/MS procedure
Authors: Hawke D. , Mazouni C. , André F. , Baggerly K. , Baggerly K. , Tsavachidis S. , Buzdar A. U. , Martin P. , Kobayashi R. , Pusztai L. 
Published: August 2020

e22072 Evaluation of serum profiles changes after neoadjuvant chemotherapy for breast cancer using MALDI-TOF / MS procedure. Background: Response to primary chemotherapy (CT) for breast cancer is heterogeneous among patients and a more tailored treatment would be beneficial in term of reducing exposure to an unnecessary toxicity and optimization of response rates. Mass spectrometry analysis of serum might be helpful in detecting specific changes in response to primary CT. Methods: An applied Biosystems 4700 Proteomics Analyzer matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometer was used. A breast cancer cohort of 78 sera samples from 39 HER2 positive patients consisting of matched pretreatment and (6 months) posttreatment samples was used. Blood samples were collected serially before each treatment cycle every 3 weeks of neoadjuvant CT. Samples were divided into those who achieved pathological complete response (pCR, n= 20) and those who had residual disease (RD, n=19). Low-mass differentially expressed peptides were identified using MALDI-TOF/TOF. Results: This procedure yielded a total of 2329 and 3152 peaks respectively, for the responders and non-responders. Biological variation analysis revealed a total of 32 peaks for responders and 643 peaks for non-responders to be differentially regulated with a false discovery rate less than 20%. A total of 8 differentially expressed proteins were identified from their peptides after digestion and LC-MALDI-TOF/TOF. Four in tumors with pCR (AFM, C3, hemopexin, SAP) and four proteins in the RD group were identified (AP1, hemopexin, Complement B, amyloid P component) Conclusions: Our study suggests that MALDI mass spectrometry may be used to predict the tumor response to neoadjuvant chemotherapy. Proteomic analysis may be useful in developing tailored chemotherapy for breast cancer. No significant financial relationships to disclose.
Correlation of WT1 expression with tumor characteristics and clinical outcomes in mucinous breast carcinoma
Authors: Liang H. , Ye F. , Lembersky B. C. , Bhargava R. 
Published: August 2020

e22075 Background: Wilms' tumor 1(WT1) protein expression has been evaluated by immunohistochemistry (IHC) in various histologies of series of invasive breast carcinomas. It is overexpressed predominantly in the mucinous subtype, in both pure and mixed mucinous carcinoma. The objective of this study was to correlate WT1 expression with the clinical and biological features of mucinous breast carcinoma as well as with the clinical outcomes and management. Methods: A total of 83 cases of pure and mixed mucinous breast carcinoma diagnosed between January 1999 and May 2006 were selected from the Magee-Womens Hospital, University of Pittsburgh Medical Center pathology registry. WT1 expression was examined by IHC using tissue microarrays (TMA). Only nuclear staining for WT1 was counted. Patient age, tumor features, management, and clinical outcomes were recorded, and survival was documented at the last patient followup. Results: Median follow-up was 59m (13–158m). The vast majority of patients had strong expression of hormone receptors. Only 3 patients had HER2-positive tumors. Disease free survival (DFS) was 92%. Overall survival (OS) was 88%. Sixty four cases had sufficient tissue on TMA for WT1 expression. Among them, WT1 expression was identified in 21/33 (64%) of pure mucinous carcinomas and 9/31 (29%) of mixed type tumors. Patients in both WT1-positive and WT1-negative groups had similar rates of breast conserving surgery, adjuvant radiotherapy and endocrine therapies. However, patients in the WT1-positive group were older (median age 74 vs 55 years, p=0.02) and had smaller tumor (median tumor size 1.2 vs 1.75 cm, p=0.03) as compared to those in WT1-negative group. More patients with WT1-negative tumors received adjuvant chemotherapy (40.6% vs 13.8%, p=0.02). Although all the recurrences were observed in patients with WT1-negative tumors, there was no statistical association of DFS or OS with WT1 expression. Conclusions: Overall, both pure and mixed mucinous breast carcinomas have excellent prognosis. This retrospective study demonstrated that WT1 expression may be associated with favorable outcome/pathologic features for patients with mucinous breast carcinoma. Prospective clinical studies will help to further elucidate these findings. No significant financial relationships to disclose.
Association of hypoxia inducible factor-1 alpha and carbonic anhydrase IX overexpression and survival in HER2/neu positive hormone-unresponsive breast cancer patients
Authors: Kaya A. O. , Gunel N. , Benekli M. , Akyurek N. , Buyukberber S. , Tatli H. , Coskun U. , Yildiz R. , Yaman E. , Ozturk B. 
Published: August 2020

e22078 Background: Hypoxia is common in solid tumors with overexpression of hypoxia inducible factor-1 alpha (HIF-1 alpha) and carbonic anhydrase IX (CA IX). In our study, we evaluated HIF-1alpha and CA IX expression in Her-2/neu positive hormone- unresponsive breast cancer patients. Methods: One hundred and eleven patients with operable breast cancer were identified who had paraffin-embedded tissue blocks available. Estrogen and progesterone receptors and Her-2/neu expression were evaluated by immunohistochemistry (IHC). FISH was used for IHC 2+ cases. We assigned two groups with 56 patients who have ER and PR receptor positive, Her-2/neu negative (group 1) and with 55 patients who have ER and PR negative, Her-2/neu positive (group 2). HIF 1 alpha and CA IX expressions were investigated and results were compared for both groups. Additionally, we investigated the association between HIF 1 alpha and CA IX expressions with stage, grade, lymph node metastasis, tumor dimension and menopause status. Results: Median age of the patients was 52 years (range, 34–77) in group 1 and 47 years (range, 27–83) in group 2. Nuclear HIF-1alpha expression was detected in 26 (46.4%) and 46 patients (83.6%) in group 1 and 2, respectively (p<0.0001). Membranous CA IX expression was detected in 25 (46.4%) and 37 (67.3%) patients in group 1 and 2, respectively (p<0.013). In group 1, median disease free survival (DFS) has not been reached. In group 2, median DFS was 46 months (95%CI: 42.98–49.02) (p= 0.0308). In group 1, median overall survival (OS) was 108 months (95%CI: 90.65–125.35) whereas in group 2, median OS was 75 months (95 % CI: 64.94- 85.06) (p= 0.0339). Conclusions: HIF-1 alpha and CA IX overexpressions were more frequently observed in ER and PR negative, Her-2/neu positive breast cancer. This was associated with shorter disease-free and overall survival. No significant financial relationships to disclose.
Profiling of receptor tyrosine kinases (RTK) activation in metastatic breast cancer lesions via proximity mediated microarray immunoassay
Authors: Kim P. , Liu L. , Liu X. , Harvie G. , Zhang L. , Kirkland R. , Magonova K. , Ybarrondo B. , Singh S. 
Published: August 2020

e22095 Background: The changes in the tumor receptor expression status between primary site and metastatic lesions are known to occur at a significant frequency (∼15–20%). Methods for profiling RTK activation patterns on metastatic tumors may provide valuable insight into the shifting disease pathogenesis allowing better selection of therapy combination for individual patient. Methods: A novel technology capable of specifically detecting phosphorylation events in ErbB family RTKs has been developed. It utilizes the formation of a unique immuno-complex requiring the co-localization of two detector enzyme-conjugated-antibodies once target proteins are captured on the microarray-surface. The channeling events between two detector enzymes (Glucose Oxidase & Horse Radish Peroxidase) in proximity enabled the profiling of the RTK with extreme sensitivity. The analytical specificity is greatly enhanced given the requirement for simultaneous binding of three different antibodies. We used 29 frozen breast cancer tissues (stage II to IV) as a model system for metastatic fine needle aspirate (mFNA) RTK profiling. Results: Out of 29 mFNA-samples, 27% (8/29) showed highly activated HER2, and 2 samples (6%) showed intermediate level of HER2-activation. Two of the 8 samples with high level HER2-activation showed significant level of HER1-activation as well. A similar level of HER1-activation was also found in one of the samples with intermediate HER2-activation. Among 19 activated-HER2-negative samples, 3 showed moderate level of HER1-activation. Conclusion: The multiplexed-proximity mediated immuno-microarray was utilized to detect the phosphorylation of HER1/HER2 in limited amount of mFNA-tissue-samples. We were able to detect activated HER1-RTKs in number of mFNA samples regardless of their HER2-IHC status. Having the ability to profile tumors at different metastatic sites with an expanded pathway panel could provide information on their differential metastatic potentials; hence minimally invasive single-passage-mFNA samples may be utilized to tailor therapy selection/adjustment and monitoring the disease progression. [Table: see text]
Treatment related changes of the serum epidermal growth factor receptor in advanced colorectal cancer
Authors: Spindler K. G. , Olsen D. , Brandslund I. , Jakobsen A. 
Published: August 2020

e22096 Background: The epidermal growth factor receptor (EGFR) is an established target for therapy in colorectal cancer. The extracellular domain of the receptor is shed into circulation and detectable by ELISA. We investigated the changes in sEGFR levels during preoperative chemoradiation (CRT) in rectal cancer patients and third-line treatment with cetuximab and irinotecan (CETIRI) in advanced disease, to elucidate the predictive or prognostic value in these settings. Methods: We included 126 healthy controls and 118 patients with chemorefractory mCRC treated with cetuximab (initial 400/m2 mg followed by weekly 250mg/m2) and irinotecan (350 mg/m2 q3w). Response was evaluated according to RECIST. Furthermore, 114 patients with locally advanced rectal tumours were treated with CRT (60 Gy/30 fractions and concomitant uftoral (300 mg/m2)/leukovorin (22.5 mg) on treatment days, followed by surgery 8 weeks post-treatment and pathological tumour regression evaluation. Pre-treatment and consecutive samples were drawn at each visit. sEGFR was measured by ELISA. Median statistics and Kaplain-Mayer curves with log-rank testing for comparison of survival rates were performed. Results: There were significant differences between the median pre-treatment sEGFR levels in controls, rectal cancer and mCRC (58 ng/ml(56–59 95% C-I), 53 ng/ml(51–55 95% C-I) and 51 ng/ml(49–53 95% C-I), respectively, p<0.000). We detected a rapid increase in sEGFR by the first on- treatment values during CETIRI (p<0.001), and a correlation between the magnitude of increase and a higher degree of skin toxicity, a well known indicator of clinical benefit to EGFR inhibitors. sEGFR in rectal cancer patients displayed a decreasing tendency during CRT (p<0.001), but no correlation to local tumour response. Patients with baseline pre-treatment level > 43.4 ng/ml (mean sEGFR of control group-2xSD) had a significantly higher OS rate than patients with low baseline levels (93% and 59% respectively, HR 0.15, P=0.002). Conclusions: We report a rapid increase in sEGFR by the onset of CETIRI, which may indicate development of skin toxicity and thereby a better change of response. Furthermore, we suggest a potential prognostic value of sEGFR measurement during CRT in locally advanced rectal cancer. No significant financial relationships to disclose.
Expression of CD133, a possible marker for cancer stem cells (CSCs), in small cell lung cancer (SCLC) cell lines and non- small cell lung cancer (NSCLC) cell lines
Authors: Hayashi T. , Tao H. , Jida M. , Kubo T. , Yamamoto H. , Otani H. , Sano Y. , Toyooka S. 
Published: August 2020

e22100 Background: Cancer stem cell (CSCs) are believed to play important roles in tumor development, recurrence or metastasis. Identification of CSCs may have a therapeutic significance. CD133 expression has been shown on a minority of various human cancer cells with high capability of self-renewal and proliferation. Therefore, CD133 is thought to be one of possible markers for CSCs. Regarding human lung cancers, the existence, prevalence or roles of CD133 positive cells has not been fully understood. Methods: We examined CD133 mRNA by quantitative real-time PCR and sorted CD133-positive cells by fluorescence-activated cell sorting (FACS) using human small cell lung cancer(SCLC) and non-small cell lung cancer (NSCLC) cell lines. We evaluated differences of cell proliferation between CD133-positive and -negative cells by MTS assay in vitro and by subcutaneous injection for non- obese diabetic/severe combined immunodeficiency (NOD/SCID) mice in vivo. Results: CD133 expression was almost restricted in SCLC cell lines. CD133 mRNA expression or CD133-positive cell population was scarcely observed in NSCLC cell lines. In two SCLC cell lines examined (NCI-H82 and NCI-H69), CD133 positive cells had higher tumorgenicity both in vivo and in vitro than NSCLC cell lines. Conclusions: The expression status of CD133 is totally different between NSCLCs and SCLCs, probably reflecting the difference of these progenitor cells. Our results indicate that CD133-positive cells in SCLC cell are responsible for tumor growth. However, in view of their wide prevalence, CD133-positive cells do not seem to be a candidate for CSCs, at least in cell lines. To investigate the molecular and functional characteristics of CD133-positive cells may lead to a new therapeutic strategy for human lung cancers, especially for SCLCs. No significant financial relationships to disclose.
Soluble vascular endothelial growth factor receptor 2 (VEGFR2): New biomarker in advanced non-small cell lung cancer (NSCLC)?
Authors: Sanmartin E. , Jantus Lewintre E. , Sirera R. , Miñana M. , Navarro A. , Cabrera A. , Blasco A. , Pla A. , Rosell R. , Camps C. 
Published: August 2020

e22108 Background: An increase in VEGF expression in tumour or some blood compartments (i.e. serum or plasma) has been found in solid tumours of various origins. Several studies have suggested that ligands and receptors of the VEGFs/VEGFR system play an important role in tumour growth and is associated with metastasis and poor prognosis. The aim of our study was to investigate the usefulness of plasmatic VEGFR2 quantification as a new biomarker in advanced NSCLC. Methods: We studied 106 healthy controls (c) and 467 advanced NSCLC patients (p) (stage IIIB and IV) treated with cisplatin and docetaxel. Blood samples were collected before chemotherapy and the plasmatic levels of the VEGFR2 were determined by ELISA. Results: In the NSCLC group, the median age was 59.9, range (31–80); 82% were males. The histological subtypes were: 31.4% squamous, 49.8% adenocarcinoma, 15.3% large cell and undifferentiated and 3.5% other. There was a significant difference in the plasmatic levels of VEGFR2 between c and p (mean± SEM): 6318±152 ng/ml and 8141± 119 ng/ml, respectively (p<0.0001). On the other hand, we found no statistical differences according to sex, histology, or stage. The area under the ROC curve was 0.743 indicating that VEGFR2 is an adequate biomarker for the discrimination between c and p. Dividing the cohort in two subgroups according to VEGFR2 levels: high (>9473,9 ng/ml) and low (≤ 9473,9 ng/ml), we found significant difference in terms of Time to Progression (TTP). Patients with higher levels of VEGFR2 had a median TTP of 204 days whereas in the group with lower expression the median was 164 days, (p= 0.039). Conclusions: In advanced NSCLC, we found higher levels of soluble VEGFR2 in p than in c. There was a correlation between higher expressions of soluble VEGFR2 with better prognosis, in terms of TTP, therefore a more thorough understanding in the role of the plasmatic quantification of this angiogenic receptor in advanced NSCLC p seems to be an important task. No significant financial relationships to disclose.
Integrating microRNA and mRNA expression profiling using a novel algorithm identified a small set of unique genes upregulated in malignant pleural mesothelioma (MPM)
Authors: Suraokar M. , Corvalan A. , Chow C. , Gazdar A. , Moran C. , Raso G. , Mehran R. , Tsao A. , Wistuba I. 
Published: August 2020

e22111 Background: We employed a global profiling strategy using miRNA microarrays in MPM cell lines and archival tumor tissue. Methods: We isolated total RNA from 4 MPM cell lines, 2 control cell lines, and 16 tissue specimens from patients with resected MPM (n=8) and normal counterpart (n=8) patients as controls. Total RNA was labeled with Cyanine 3, then hybridized with Agilent human miRNA microarray v1 slides. Results: Preliminary miRNA profiles show up-regulation of 44 versus down-regulation of 29 miRNA's in MSTO-211H cancer cells compared to HCT-4012 (pleural telomerase-transformed control). Profiling of 16 tissue specimens (8 normal vs 8 MPM) revealed down-regulation of 11 miRNA's in MPM tumor tissue. To focus on relevant miRNA that regulate genes involved in carcinogenesis and progression, we identified > 1000 unique genes using the online targetscan 4.2 program ( http://www.targetscan.org ), which predicts biological targets of miRNAs by identifying the presence of conserved 8-mer and 7-mer sites that match the seed region of each miRNA. We then explored a novel screening strategy, which combines mRNA expression dataset with the miRNA dataset, to narrow the list of relevant miRNA's. We conducted gene expression profiling on the cell lines and MPM tissue samples with Affymetrix U133 plus 2.0 chips. Bioinformatic analysis was conducted with MeV: MultiExperiment Viewer software, data reduction techniques (Correspondance Analysis), hierarchical clustering methods, and Serial Analysis for Microarray (SAM), and showed up-regulation of ∼300 genes in MPM compared to normal tissues. We then computed of the ∼300 mRNA's up-regulated in MPM only 32 are recognized by the 11 down-regulated miRNA's using the targetscan 4.2 algorithm. Most of the miRNA's regulate single messages while ∼20 % of the messages are regulated by more than 1 miRNA's. Some of these targets include Ets variant 1 and Protein kinase C - epsilon. Conclusions: This innovative approach of selecting highly relevant miRNA is feasible and enables discovery of novel genes based on their ability to be bound by single or multiple miRNA's. Validation of our profiling studies using real-time PCR and protein analysis methods will be presented. No significant financial relationships to disclose.