bluuebunny/crossref_metadata_2025_split huggingface dataset:
Molecular characterization of circulating tumor cells in human metastatic colorectal cancer.
Authors: Barbazan Jorge , Alonso-Alconada Lorena , Muinelo-Romay Laura , Vieito Maria , Abalo Alicia , Alonso-Nocelo Marta , Candamio Sonia , Vazquez Francisca , Vidal Yolanda , Fernandez Beatriz , Abdulkader Ihab , Casares Maria de los Angeles , Gomez-Tato Antonio , Abal Miguel , Lopez Rafael 
Published: March 2020

10534 Background: Metastatic colorectal cancer (mCRC) relies on the detachment of aggressive malignant cells from the primary tumor into the bloodstream, being this Circulating Tumor Cells (CTC) the principal source of the further metastasis. Clinically, the presence of CTC is associated with poor prognosis and there exists a clear necessity for more specific and efficient chemotherapies in the treatment of mCRC. Our aim was to overcome this adverse scenario through the massive molecular profiling of the CTC population isolated from mCRC patients. Methods: CTC’s were magnetically isolated by using anti-EpCAM coupled magnetic beads from 6 mCRC patients and 3 healthy controls. The presence of CTCs was evaluated by citokeratins 8, 18 and 19 staining. RNA from CTCs was amplified by a whole transcriptome amplification system (WTA) and resulting material was hybridized onto gene expression arrays. Bioinformatics were used to array analysis. Selected genes were validated by RT-qPCR. Results: 410 transcripts were found to specifically characterise the CTC population after array signals comparison between mCRC patients and controls. Gene-gene interaction analysis generated networks related with cell migration, adhesion or apoptosis resistance. Gene ontology revealed similar functions. Signalling pathways such as RhoA, PKA, ILK, integrins or actin cytoskeleton signalling were found as relevant in CTCs. Eleven genes (TGFB1, APP, TIMP1, CD9, CLU, ITGB5, LIMS1, RSU1, VCL, BMP6 and TLN1) were validated by real-time PCR in 20 mCRC patient and 10 control samples. All genes showed a significantly higher expression in patients (p<0.0001). Except from TLN1 and CD9, all genes had a prognostic value in terms of progression free survival (p<0.05). Conclusions: We described the molecular profiling of CTCs in stage IV CRC patients, characterized by a migratory and invasive phenotype. Specific and sensitive diagnostic/prognostic markers were obtained. Our strategy represents an innovative and promising approach in the clinical management of mCRC patients.
A new medical device for in vivo capturing of circulating tumor cells in breast cancer (BC) patients.
Authors: Murawa Dawid , Herold Stefanie , Kim Phillip Sangwook , Schmitz Arndt , Krahn Thomas , Murawa Pawel , Zabel Maciej , Morgenthaler Nils G. , Nowaczyk Piotr , Luecke Klaus 
Published: March 2020

10537 Background: In BC, the number of circulating tumor cells (CTCs) is discussed as a prognostic and stratification biomarker, and could also reflect treatment efficacy. Currently, CTCs are isolated ex vivo from a small volume of blood. Results from a larger volume of blood are scarce. The aim of the study was to assess a functionalized and structured medical wire (FSMW) for in vivo capturing of CTCs directly from the blood stream of BC patients. Methods: The device was inserted in a cubital vein through a standard cannula for thirty minutes. The interaction of target CTCs with the FSMW was mediated by antibodies directed against the epithelial cell adhesion molecule (EpCAM). To confirm binding of CTCs to the wire, the immunohistochemical positive staining against EpCAM as well as negative staining for CD45 was performed. There were 54 applications of the wire in 42 stage I-IV BC patients (12 double applications). Enumeration data from 37 BC patients with 49 applications (5 failed subsequent analyses) were assessed. CTC counts on 23 devices were directly compared to counts by CellSearch. Results: The device was well tolerated in all 54 applications without side effects. We obtained in vivo isolation of CTCs in 44 of 49 applications to BC patients (89.7 %). The sensitivity was similar for early and late stage BC patients. The median (range) of isolated EpCAM-positive CTCs was 5 (0-515). The CellSearch method reached a sensitivity of 18.5%. In all paired samples the number of CTCs detected with the FSMW was higher or equal to CellSearch, regardless of the disease stage. Linear regression of the data of the double application of the FSMW showed a very good concordance (r2 = 0.97, p<0.0001). Conclusions: Whilst well tolerated without side effects, the CTC detection rate of the FSMW in BC patients was nearly 90 %. CTC detection was obtained in 18.5% by the CellSearch. Double application of FSMWs in the same patient indicates ample precision. This proof of concept study may have important clinical implications, as the device may improve early detection, prognosis and therapy monitoring of BC patients. The molecular analysis of the captured CTCs could become a breakthrough in personalized medicine.
Different EGFR/KRAS mutation spectrums in lung cancer between never smokers and heavy smokers.
Authors: Takamochi Kazuya , Oh Shiaki , Suzuki Kenji 
Published: March 2020

10548 Background: EGFR mutations are more commonly found in lung adenocarcinomas in never smokers, while KRAS mutations are more frequently present in those in heavy smokers. Although the incidence is low, EGFR mutations are also found in tumors in heavy smokers and KRAS mutations also occur in tumors in never smokers. Therefore, there may be different biological characteristics in EGFR/ KRAS mutated lung cancers according to the smoking status. Methods: This was a retrospective review of 382 patients with surgically resected lung cancers between February 2009 and March 2011. The clinicopathological factors (age, gender, histology, serum CEA level, pathological nodal status, lymphatic permeation, and blood vessel invasion) and EGFR/KRAS mutation spectrums were compared between never smokers (pack-year = 0, n = 161) and heavy smokers (pack-year > 20, n = 167). Results: EGFR mutations were detected in 88 (55%) never smokers and in 33 (20%) heavy smokers. In contrast, K-ras mutations were detected in 7 (4%) never smokers and in 31 (19%) heavy smokers. Both EGFR and KRAS mutations were more frequently observed in female never smokers and in male heavy smokers. The spectrum of both EGFR and KRAS mutation differed according to the smoking status. Minor EGFR mutations other than exon 21 L858R and exon 19 deletions that are activating mutations indicating the efficacy of EGFR tyrosine kinase inhibitors (TKIs) were more frequently found in heavy smokers than in never smokers. G→A transitions were more common KRAS mutations in never smokers and G→T transversions that are thought to be related to exposure to the polycyclic aromatic hydrocarbons found in cigarette smoke were more common KRAS mutations in heavy smokers. Conclusions: The EGFR/KRAS mutation spectrums in lung cancer were quite different in never smokers and heavy smokers. Further study is needed to evaluate the relationship between the efficacy of such molecular targeting agents as EGFR TKIs and the EGFR/KRAS mutation spectrums.
Prediction of late metastasis in node-negative breast cancer.
Authors: Schmidt Marcus , Hellwig Birte , Gehrmann Mathias , Koelbl Heinz , Boehm Daniel , Cadenas Christina , Rahnenfuehrer Joerg , Hengstler Jan G 
Published: March 2020

10551 Background: Prediction of late metastasis is of clinical relevance in breast cancer. However, systematic genome-wide studies to identify genes associated with increased risk of metastasis 5 or more years after surgery are scarce. Methods: We examined the natural course of disease in three previously published cohorts (Mainz, Rotterdam, Transbig) including 766 node-negative breast cancer patients with gene array data who did not receive systemic chemotherapy in the adjuvant setting. We established a Cox regression based method adjusted for multiple testing that identified genes predicting late metastasis (5 or more years after surgery). Only those genes were accepted that showed similar results in all three cohorts. Metastasis-free survival (MFS) was analyzed with univariate and multivariate Cox regression. Results: We identified 9 genes [ABCC5 (Hazard Ratio (HR) 2.19, p=0.003), EDDM3B (HR 3.58, p=0.044), RAD23B (HR 0.37, p<0.001), XYLT2 (HR 2.19, p=0.027), DDX18 (HR 0.35, p=0.006), GPBP1L1 (HR 0.20, p=0.018), UBB (HR 9.73, p=0.025), RPS24 (HR 0.20, p=0.050), GPC1 (HR 2.36, p=0.013)] predicting late metastasis. These genes retained their independent prognostic significance after adjustment for established clinical factors (age, tumor size, grade, hormone receptor status, HER2) and biological motives like estrogen receptor, proliferation, B or T cells. These late-type genes are largely associated with resistance to hypoxia, apoptosis and DNA damage, suggesting that they might contribute to persistence of disseminated tumor cells. Conclusions: Genes associated with late metastasis offer a perspective to identify breast cancer patients suitable for additional and prolonged therapies.
Central review of discordant samples for microarray based on ER, PR, and HER2 and local IHC/FISH assessment worldwide from 827 patients.
Authors: Wesseling Jelle , Tinterri Corrado , Sapino Anna , Zanconati Fabrizio , Lutke Holzik Martijn , Nguyen Bichlien , Deck Kenneth B. , Querzoli Patrizia , Perin Tiziana , Giardina C , Seitz Gerhard , Guinebretiere Jean-Marc , Barone Julie , Watanabe Toru , De Snoo Femke , Stork-Sloots Lisette , Cusumano Pino 
Published: March 2020

10554 Background: Differences in fixation and IHC and subjective interpretation can substantially affect the accuracy and reproducibility of estrogen receptor (ER), progesterone receptor (PR) and HER2 expression. The commercially available TargetPrint test measures the mRNA expression level of ER, PR and HER2 and is 98% concordant with centrally assessed ER as presented by Viale et al, SABCS 2011. This study compares results from the microarray-based TargetPrint with IHC and FISH (for HER2 IHC2+) generated by local standard procedures. Methods: Fresh tumor samples (core needle biopsies or surgical) were collected for 831 patients diagnosed with breast cancer stage I to IV (Feb 2008 - Jan 2011) from 22 hospitals from Europe, New Zealand, Japan and US. The results of the IHC/FISH assessments performed according to the local standards at the hospitals were compared to the quantitative gene expression readouts with TargetPrint. Discordant cases were centrally reviewed for IHC/FISH assessment. Results: Of the 831 samples, IHC assessment was unknown for 4 ER/ PR samples; HER2 was unknown for 12 samples. Comparison of IHC and gene expression read out by TargetPrint showed a concordance of 95% for ER; 83% for PR and 94% for HER2. In this study, 3% of all IHC ER positive samples were classified negative by microarray, and 11% of IHC PR positive samples were classified negative by microarray. For HER2, 4% of IHC/FISH HER2 positive samples were classified negative by microarray and 2% of IHC/FISH HER2 negative samples were classified positive by microarray. Most notably, all available 5 ER IHC negative/TargetPrint positive samples turned out to be positive with central re-assessment. HER2 IHC2+ samples with discordant classifications for TargetPrint and local assessment are currently being reviewed for FISH/SISH assessment. Conclusions: Microarray based readout of ER, PR and HER2 status using TargetPrint is fairly comparable to local IHC and FISH analysis in 827 analyzed samples in various hospitals worldwide. However, re-assessment of discordant cases –especially IHC ER-/TargetPrint ER+ cases- confirms TargetPrint to be a useful high quality second opinion for local IHC/FISH assessment.
Lymphopenia combined with low TCR diversity to predict overall survival in metastatic breast cancer patients.
Authors: Manuel Manuarii , Tredan Olivier , Bachelot Thomas , Clapisson Gilles , Courtier Anais , Parmentier Gilles , Grives Audrey , Perez Solene , Mouret Jean-Francois , Perol David , Chabaud Sylvie , Ray-Coquard Isabelle , Labidi-Galy Sana Intidhar , Heudel Pierre , Pierga Jean-Yves , Caux Christophe , Blay Jean-Yves , Pasqual Nicolas , Menetrier-Caux Christine 
Published: March 2020

10562 Background: Lymphopenia (<1 Giga/L) before initiation of chemotherapy is a predictive factor for toxicity and death in metastatic phase for cancer patients. Combinatorial diversity of T Cell Receptor beta chain (TCR-ß), as a measure of T cell repertoire diversity, was investigated and tested either alone or in combination with lymphopenia as a prognostic factor for overall survival (OS) in first line treatment of metastatic breast cancer (MBC) patients. Methods: Using semi quantitative multiplex PCR, the V-D-J combinatorial diversity of the TCR was measured on cryo-preserved blood samples from 2 cohorts of MBC patients before the initiation of the first line chemotherapy: in an experimental cohort (cohort A, n=66) and in a validation series (cohort B, n=67). A prognostic score, defined NDL (Number & Diversity of Lymphocytes) combining lymphocyte count and TCR diversity was delineated. Univariate and multivariate analysis of prognostic factors for OS were performed in both cohorts. Results: Lymphopenia (<1 Giga/L) was associated with shorter OS for cohort B while TCR diversity ≤33% (called divpenia) was associated with a reduced OS in cohort A. The combination of lymphopenia with low TCR diversity (called lympho-divpenia) was associated with poor OS compared to patients with either lymphocyte count ≥1 Giga/L or diversity >33% or both, in cohort A (median OS: 7.6 vs 24.5 months, p.value =0.0006) and cohort B (median OS 10.6 vs 22.9 months, p.value =0.0035). In a multivariate analysis, including all significant clinical factors from the univariate analysis (PS, liver metastasis, hemoglobin) lympho-divpenia was found to be an independent prognostic factor in the pooled cohort (A+B) (p=0.005) along with triple negative tumors (p=0.011) and hemoglobin level (11.5 g/dL) (p=0.009). Conclusions: NDL score combining lymphopenia and reduced TCR diversity seems to be a strong prognostic factor for OS and could be use to improve care quality of MBC patients.
Detection of EML4-ALK in serum RNA from lung cancer patients using MassARRAY platform.
Authors: Kudo Keita , Nishio Makoto , Sakai Kazuko , Tanimoto Azusa , Sakatani Toshio , Saito Ryota , Kaburaki Kyohei , Yanagitani Noriko , Horiike Atsushi , Ohyanagi Fumiyoshi , Nishio Kazuto 
Published: March 2020

10569 Background: The presence of the transforming fusion gene EML4-ALK in non–small cell lung cancer (NSCLC) is a predictive marker for the efficacy of ALK kinase inhibitors. The break apart fluorescence in situ hybridization (FISH) assay has served as the standard test for identification of NSCLC patients with ALK gene rearrangement. Circulating RNA in plasma/serum is an emerging field for noninvasive molecular diagnosis. We previously developed a MassARRAY assay to detect nine types of EML4-ALK fusion gene (variants 1, 2, 3a, 3b, 4, 5a, 5b, 6, and 7). The assay is based upon region-specific PCR followed by specific single-base primer extension reactions; the extended products are resolved using MALDI-TOF mass spectrometry. In this study, we evaluated whether EML4-ALK fusion RNA could be detected in serum obtained from NSCLC patients. Methods: The RNA was extracted from serum obtained from 12 lung cancer patients with ALK rearrangements by FISH analysis. The extracted RNA was reverse transcribed into cDNA, and the resultant cDNA was amplified by nested PCR. The PCR product was subjected to MassARRAY assay to identify EML4-ALK variants. Results: The EML4-ALK RNA (variant 1 and 3) was detected in serum from 2 of 12 patients. These are confirmed by subcloning and sequencing. Conclusions: The detection of EML4-ALK in serum RNA samples by MassARRAY assay is feasible. This noninvasive assay will be useful in patients with insufficient or unavailable tumor specimens. Additional experiments will reveal the sensitivity and specificity of this detection system.
The effect on detection rate of second cancers after the addition of MRI to conventional screening mammography in patients with a history of breast cancer.
Authors: Weinstock Chana , Campassi Cristina , Goloubeva Olga G. , Chumsri Saranya , Bao Ting , Kesmodel Susan , Feigenberg Steven J. , Wooten Kathleen , Ioffe Olga B. , Tkaczuk Katherine Hanna 
Published: March 2020

10572 Background: Mammography is currently used in the surveillance of breast cancer survivors, who are at increased risk of developing ipsilateral and contralateral breast cancers regardless of age at diagnosis or time since diagnosis. Several prospective studies have shown the utility of breast MRI in other high risk populations; however, little data exists on the use of MRI for surveillance of breast cancer survivors. We aimed to compare the outcome of surveillance breast MRI vs. mammography in this population. Methods: We identified women <65 with non-metastatic breast cancer or DCIS with at least one MRI performed at our center >11 months after initial diagnosis, along with a mammogram done within 6 months of the MRI. We compared the outcome of MRI and mammography in terms of biopsies performed as well as in detection of new cancers. Results: Of 512 consecutive charts reviewed, 204 patients met inclusion criteria, 105 (51.4%) of whom were African-American. The average number of MRIs per patient was 2.3 (range 2-7), with a total of 474 MRIs performed between 2005 and 2011. MRI resulted in BIRADS scores of 1 or 2 in 73.5% of studies vs. 84.4% for mammography. There were 19 biopsies performed due to MRI findings alone, 7 done due to mammographic findings alone, and 6 biopsies done based on abnormalities seen on both MRI and mammography. There were 7 malignancies identified based on an abnormal MRI, 3 seen on both MRI and mammography, and none identified via mammography alone. The malignancies identified via MRI alone included 1 patient with DCIS, 5 with stage I disease, and 1 with isolated lung metastases. Of the 10 recurrences detected, 5 (50%) were in African Americans. Two patients developed interval cancers within 6 months of normal screening MRI and mammography. Sensitivity and specificity for MRI were 83.3% (95% CI 0.51-0.97) and 92.2% (95% CI 0.87-0.95), vs. 25% (95% CI 0.05-0.57) and 94.8% (95% CI 0.90-0.97) for mammography. Positive and negative predictive values were 40% and 98.9% for MRI vs. 25% and 95.2% for mammography. Conclusions: Gadolinium-enhanced breast MRI is a useful surveillance modality in breast cancer survivors < age 65. Prospective studies are needed in this population.
Correlation of ERCC1 expression on circulating tumor cells with progression-free survival in metastatic non-small cell lung cancer patients treated with platinum-based chemotherapy.
Authors: Riess Jonathan , Das Millie Snigdha , Frankel Paul Henry , Schwartz Erich , Bennis Robyn , Hsieh H Ben , Liu Xiaohe , Ly Janey , Zhou Lisa Ya , Nieva Jorge J. , Bruce Richard , Wakelee Heather A. 
Published: March 2020

10574 Background: Biomarkers predicting efficacy of chemotherapy are highly desirable. Fiber array scanning technology (FAST) is a novel method of detecting circulating tumor cells (CTCs) that does not employ an EpCam enrichment step. The purpose of this study was to use FAST to evaluate ERCC1 expression on CTCs to determine whether ERCC1 expression correlates with progression-free survival (PFS) in patients who received platinum-based chemotherapy for metastatic non-small-cell lung cancer (NSCLC). Methods: Peripheral blood from one hundred enrolled patients with metastatic NSCLC was collected by two institutions (Stanford Cancer Institute and Billings Clinic Cancer Center). FAST was used to identify individual CTCs on immunofluorescence by pancytokeratin antibodies. Nuclear localization of ERCC1 expression by immunofluorescence was quantified on individual CTCs. Total patient ERCC1 levels were determined from the average expression of all the CTCs in each patient sample. Fifty-seven of the one hundred patients enrolled received platinum chemotherapy. Seventeen of those fifty-seven patients (30%) had ≥ 2 evaluable intact CTCs and were analyzed retrospectively. Linear regression (F-test) was used to evaluate the correlation between ERCC1 expression and PFS. Kaplan-Meier survival analysis (log-rank test) was used to compare PFS in patients with CTCs with no detectable ERCC1 expression versus patients with CTCs that expressed any level of ERCC1. Results: PFS decreased with increasing ERCC1 expression (P<0.04 F-test, linear regression). Lack of ERCC1 expression was associated with longer PFS (266 days vs. 172 days, log-rank test P<0.02). The difference in survival was statistically significant with a hazard ratio of 4.20 (95% CI 1.25-14.1, p<0.02, log-rank test). Conclusions: In this small study, using FAST to isolate CTCs, low expression of ERCC1 on evaluable CTCs correlated with increased PFS in patients with metastatic NSCLC who received platinum chemotherapy. A larger, prospective study to validate these retrospective results is warranted.
Use of early 18f-fluorodeoxyglucose-positron emission tomography to predict clinical outcome of patients with advanced non-small cell lung cancer on gefitinib treatment versus carboplatin plus paclitaxel.
Authors: Kanazu Masaki , Maruyama Kaoru , Ando Masahiko , Asami Kazuhiro , Ishii Mari , Minomo Shojiro , Matsuda Yoshinobu , Kawaguchi Tomoya , Atagi Shinji , Kusunoki Yoko , Kubo Akihito , Takada Minoru 
Published: March 2020

10577 Background: Early prediction of clinical efficacy is of great value in cancer patients in avoiding unnecessary toxicities and giving them another chance for different treatments. This study aimed to assess the 18F-fluorodeoxyglucose-positron emission tomography (FDG-PET) at 3 days on treatment as an early predictor of clinical outcome in patients with advanced non-small cell lung cancer (NSCLC) treated with gefitinib and in those treated with cytotoxic chemotherapy. Methods: This study comprised two groups: patients with stage IIIB or IV NSCLC were treated with gefitinib (250mg) once daily (gefitinib group) or with carboplatin (AUC 6, day 1) plus paclitaxel (200mg/m2, day 1) q21 days (CP group) according to the physicians’ choice. FDG-PET was performed before, 3 days on and 28 days on each treatment. Reduction of tumor FDG uptake was assessed by using standardized uptake value (SUV). Metabolic response was defined as reduction of FDG uptake in the tumor ≥ 25% according to the criteria of the European Organization for Research and Treatment of Cancer. Metabolic response was correlated with clinical outcomes. Results: This study included 38 patients: 19 in gefitinib group and 19 in CP group. Reduction of SUV at 3days on treatment preceded tumor shrinkage more closely in gefitinib group. Metabolic response was significantly correlated with longer progression-free survival (PFS) in gefitinib cohort (median PFS 15.8 months [95% CI 13.2-18.4] vs. 3.7 months [95% CI 0.1-8.0], p < 0.001), but not in CP group (median PFS 5.7 months vs. 2.9 months, p = 0.054). Furthermore, metabolic response was significantly correlated with longer overall survival (OS) in gefitinib group (median OS 28.7 months [95%CI 23.5-33.9] vs. 9.8 months [95% CI 2.1-17.5], p = 0.009), but not in CP group (median OS 13.9 months vs. 10.5 months, p = 0.56). In multivariate analysis using Cox hazards models, metabolic response was a significant predictive factor of PFS and OS. Conclusions: Reduction of SUV levels on FDG-PET at 3 days on treatment may predict response and survival in gefitinib-treated patients with advanced NSCLC.