bluuebunny/crossref_metadata_2025_split huggingface dataset:
Everolimus for postmenopausal women with advanced breast cancer: Updated results of the BOLERO-2 phase III trial.
Authors: Arena Francis P. , Noguchi Shinzaburo , Pritchard Kathleen I. , Burris Howard A. , Rugo Hope S. , Gnant Michael , Hortobagyi Gabriel N. , Latini Luciano , Yardley Denise Aysel , Melichar Bohuslav , Petrakova Katarina , Harb Wael , Feng Wentao , Cahana Ayelet , Taran Tanya , Campone Mario , Baselga José , Sahmoud Tarek , Lebwohl David Edward , Piccart-Gebhart Martine J. 
Published: February 2017

99 Background: Current treatment options for postmenopausal patients with estrogen-receptor–positive breast cancer (BC) who relapse or progress on a nonsteroidal aromatase inhibitor (NSAI) are limited. The BOLERO-2 trial supports the activity of everolimus (EVE; an oral mammalian target of rapamycin [mTOR] inhibitor) added to the steroidal aromatase inhibitor exemestane (EXE) to prolong progression-free survival (PFS) in this patient population. Long-term PFS and survival data are awaited. Methods: BOLERO-2 is a phase 3, double-blind, randomized, international trial comparing EVE (10 mg once daily) + EXE (25 mg once daily) vs. placebo (PBO) + EXE in postmenopausal women with advanced estrogen-receptor–positive BC progressing or recurring after NSAIs (letrozole or anastrozole). Patients were randomized (2:1) to EVE + EXE or PBO + EXE. The primary endpoint was PFS by local investigator assessment. Main secondary endpoints included centrally assessed PFS, overall survival (OS), safety, bone turnover, and overall response rate. Results: Baseline disease characteristics including tumor burden and prior cancer therapy were well balanced between treatment arms (N = 724). Median PFS was doubled and response rates were consistently improved with EVE + EXE (n = 485) vs PBO + EXE (n = 239) in interim analyses. Median PFS by local assessment was ~3 mo with PBO + EXE vs 6.9 mo (hazard ratio [HR], 0.43; P < .0001) and 7.4 mo (HR, 0.44; P < .0001) with EVE + EXE at 7.5 mo and 12.5 mo follow-up, respectively. Fewer deaths were reported with EVE + EXE (17.2%) vs PBO + EXE (22.7%) at 12.5 mo follow-up. Safety profiles were consistent with previous reports for mTOR inhibitors. PFS data including 528 events (protocol-specified final analysis), and updated OS and safety data will be presented. Conclusions: Adding EVE to EXE markedly prolonged PFS in patients with NSAI-refractory advanced estrogen-receptor–positive BC. There were fewer deaths among patients receiving EVE, and further follow-up will evaluate the effect of EVE on OS.
Comparison of COBAS 4800 KRAS, PCR TaqMan, and PCR high-resolution melting assays for the detection of KRAS somatic mutations in formalin-fixed paraffin-embedded colorectal tumors.
Authors: Harle Alexandre , Merlin Jean-Louis 
Published: February 2017

103 Background: Colorectal cancer (CRC) is the 2nd most common cancer with more than one million new cases diagnosed every year. From 2006 to 2008, several studies showed the importance of the KRAS oncogene in the treatment of metastatic CRC (mCRC) and response to anti-EGFR therapies like cetuximab or panitumumab. KRAS is a small G protein which can bear activating mutations in 40% cases of mCRC. Numerous methods have been described for the detection of KRAS mutations in FFPE tissues. Direct sequencing is the gold standard but the most sensitive methods are real time PCR assays based. The new COBAS4800 KRAS developed by Roche Diagnostics is a CE IVD validated assay using real time PCR and TaqMelt technology. This assay is validated for the detection of 19 KRAS somatic mutations on exon 2 (codons 12/13) and exon 3 (codon 61) in specimen containing at least 5% of mutated tumoral cells. Methods: We compared COBAS with previously validated PCR TaqMan and PCR High Resolution Melting (HRM) assays. The PCR TaqMan and HRM are a real time PCR based assays. TaqMan can detect accurately 0.5 to 1% of mutated tumoral DNA on codon 12 (G12A, G12C, G12D, G12R, G12S and G12V) and on codon 13 (G13D). HRM can detect 5 to 10% of tumoral DNA on codon 12 and 13. 80 FFPE samples of colorectal tumors resections or biopsies have been collected and processed with the 3 assays. Results have been compared using kappa statistics. Results: On 80 samples, 69 were interpretable with COBAS and TaqMan and 62 with COBAS and HRM. No statistically significant difference between COBAS and TaqMan was found (k=0.942 ; p<0.001). Only 2 discordant cases were found mainly regarding codon 61 mutations not detected with TaqMan. No statistically significant difference between COBAS and HRM was found (k=0.903 ; p<0.001). Three discordant cases were found two regarding codon 61, not detected with HRM and one G12S mutation (detected by COBAS and TaqMan). Conclusions: The 3 assays can detect accurately KRAS mutations. Less ininterpretable results were found with COBAS and TaqMan assays. A pre screening with COBAS and mutation characterization with TaqMan seem to be a good scheme for routine diagnostic of KRAS mutations.
Validation of galectin-7 using backscattering interferometry.
Authors: Olmsted Ian Roys , Massion Pierre P. , Bornhop Darryl J. , Hassanein Mohamed , Hoeksema Megan 
Published: February 2017

15 Background: More than 60% of lung cancer patients are diagnosed with advanced disease due to a lack of early diagnosis tools. These patients are ineligible for surgical resection and have a poor prognosis. This situation could be avoided if we could quantify lung cancer specific biomarkers at sufficiently low concentrations, providing an indication of disease threat. Backscattering Inerferometry (BSI) is a label-free sensor with a simple optical train that is used to quantify biomarkers in complex matrices at picomolar to femtomolar levels. Here we demonstrate that BSI can enable lung cancer biomarker validation in complex, volume constrained samples and at detection limits significantly better than ELISA. Methods: A BSI dose response curve for Galectin-7 was constructed by incubating increasing concentrations (0-20 ng/mL) of recombinant galectin-7 with 100 ng/mL of polyclonal antibody. The samples were mixed at 300 RPM for 2 before being injected into the BSI instrument and measured in triplicate each day. All reagents used were obtained from a commercial ELISA kit. An ELISA dose response curve was similarly constructed using spiked serum and spiked plasma according to the manufacturer’s recommended procedure. Finally, 9 patient serum samples were quantified using BSI and ELISA for direct comparison of the two technologies. Results: BSI was used to quantify galectin-7 in spiked serum and patients samples. The lower limit of detection with standards was determined to be 0.5 ng/mL for ELISA in serum, 10 ng/mL for ELISA in plasma, and 0.04 ng/mL for BSI in serum. In the analysis of 9 patient serum samples, all were quantifiable using BSI, which enjoys low pg/mL detection limits, yet with ELISA only 5 samples contained a galectin-7 concentration high enough to be measured. In this small sample set there was a good correlation between disease state and galectin-7 concentration as measured by BSI. Conclusions: This study demonstrates that BSI is well-suited for biomarker detection and validation, having a greater dynamic operating range and much lower limits of detection than standard commercially available ELISA kits. Future work will focus on detecting other NSCLC biomarkers that are undetectable with currently available technology.
Do insulin-like growth factor associated proteins qualify as a tumor marker?
Authors: Matuschek Christiane , Peiper Matthias , Budach Wilfried , Gerber Peter Arne , Bojar Hans , Steinbach Gerald , Rudoy Michael , Knoefel Wolfram Trudo , Schauer Matthias 
Published: February 2017

18 Background: Insulin-like growth factor (IGF)-1, -2 and insulin-like growth factor binding proteins (IGFBP) are involved in the proliferation and differentiation of cells. It has never been evaluated if the IGF-system can serve as a tumor marker in neoplasms. Methods: In our prospective study, 163 patients with colorectal cancer (22), prostate cancer (21), glioblastoma (12), head and neck tumors (17), lymphomas (20), lung cancer (34), and other entities (37) were analysed for their IGF and IGFBP serum levels at the beginning and the end of radiotherapy and compared with 13 healthy people. Subgroups of patients with local tumor disease versus metastatic disease, primary and recurrent therapy and curative versus palliative therapy were compared. Results: The serum levels of IGF-2 were significantly elevated in patients with prostate and colorectal cancer. However, sensitivity and specificity were only 70%. IGFBP-2 serum levels were elevated in patients with head and neck tumors. Again, sensitivity and specificity were only 73%. A difference between local disease and metastatic disease could not be found. A difference between IGF serum levels before and after radiotherapy could not be detected. Conclusions: The IGF-system cannot serve as a new tumor marker. The detected differences are very small and sensitivity and specificity are too low. IGF measurement is not useful for the evaluation of the success of radiotherapy in malignancies.
Growth of tumorspheres from peripherally circulating tumor cells.
Authors: Zimon Dorothea , Pizon Monica , Pachmann Ulrich A. , Pachmann Katharina 
Published: February 2017

2 Background: Even after the complete resection of a malignant tumor the menace remains, that cells that have dissociated from the tumor might resettle in distant organs and grow into life-threatening metastases. Therefore we have developed an approach (maintrac) to detect and monitor the circulating tumor suspect epithelial cells in blood. However, the question arises whether these cells have the potential to grow into metastases. Methods: Using a nondissipative approach with only one enrichment step of red blood cell lysis, the cells from the pellet, containing the white blood cells together with the putative tumor cells were cultured under conditions favoring the growth of epithelial cells. At 7, 14, and 21 days the cultured cells were stained with anti-EpCAM antibody and inspected for the appearance of epithelial antigen positive spheroids. Results: Peripherally circulating cells from patients with malignant tumors in different stages of disease were analyzed for the presence of circulating epithelial tumor suspect cells and the frequencies of tumorspheres. Tumorspheres could so far be grown from all of 20 patients in whom more than 2000/ml epithelial tumor suspect cells were detected. Fractions of tumorspheres varied from less than 1% of the epithelial tumor suspect cells in patients with primary breast cancer to up to 70% in a patient with an aggressively growing metastatic colon cancer. The size of the spheres increased from day 7 to day 21 but already had reached the size of day 21 colonies after seven days in the patient with the metastatic colon cancer. Conclusions: Here, we demonstrate that the tumor suspect cells, detected in our approach contain a subpopulation with stem cell-like properties capable of growing into tumorspheres. The frequency and growth potential of cells capable of forming spheres seems to be dependent from the properties of the primary tumor. The possibility to grow tumorspheres from peripherally circulating tumor cells may open up a new field, where the relevant cells with stem cell properties from individual patients can now be specifically analysed further for genetic endowment, transcriptional activity, heterogeneity, and stem cell markers.
PTPRF expression as prognostic/predictive marker for treatment with erlotinib in non-small cell lung cancer (NSCLC)?
Authors: Klughammer Barbara , Bordogna Walter , Delmar Paul 
Published: February 2017

22 Background: Erlotinib is an EGFR TKI established as maintenance, second- and third-line treatment in unselected NSCLC patients and as first-line treatment in patients harboring EGFR mutations. Aberrations in the receptor type protein tyrosine phosphatase (PTP) gene family were described by Jacob and Motiwala (2005) as a hallmark of carcinogenesis. Recent research established the role of PTPRS, a member of the same gene family, as necessary for the dephosphorylation of EGFR in head and neck cancer (Morris and Bivona. 2011). The role of PTPRF as an interesting prognostic marker in NSCLC and potential predictive marker for treatment with erlotinib is reported here. Methods: MERIT was a single arm phase II clinical trial (Tan et al. 2010) where all patients had biopsies performed by bronchoscopy before starting treatment with erlotinib (150 mg/day p.o.). Biopsies were snap frozen and RNA was extracted using laser capture microdissection to enrich for tumor content. Affymetrix Human U133A microarrays were used to analyse gene expression profiles. Exploratory statistical methods identified several gene candidates by comparing benefitting and nonbenefitting patients including PTPRF. These findings were confirmed by qRT-PCR in both fresh and FFPE tissue. Here we describe analysis of PTPRF in FFPE tissue from the randomized phase III SATURN trial (Cappuzzo et al. 2010) Results: PTPRF expression was found to be greater in benefitting patients than in nonbenefitting patients in MERIT HR 0.50 (0.34 - 0.73), p = 0.0004 on a continuous log2 scale. Results from SATURN further suggested predictive characteristics for PTPRF, with a larger treatment effect observed in patients with high levels as defined above median. Conclusions: Data from MERIT and SATURN suggest PTPRF to be a possibly prognostic marker for OS in NSCLC and potential predictive marker for treatment with erlotinib. This gene family is very interesting and further research, especially MoA is warranted. [Table: see text]
Response rates in patients with relapsed/refractory acute myeloid leukemia with FLT3-ITD mutation using 5-azacitadine plus sorafenib.
Authors: Alattar Mona Lisa , Kantarjian Hagop , Levis Mark J. , Richie Mary Ann , Daver Naval Guastad , Garcia-Manero Guillermo , Faderl Stefan , Borthakur Gautam , Burger Jan Andreas , Kadia Tapan M. , Grunwald Michael Richard , Dellasala Sara , Cortes Jorge E. , Ravandi Farhad 
Published: February 2017

24 Background: Outcome of patients with relapsed acute myeloid leukemia (AML) with FLT3-ITD mutation is poor. Elevated FLT3 ligand levels (FL) secondary to cytotoxic chemotherapy is a putative mechanism of resistance to the FLT3 kinase inhibitors. We hypothesized that FL levels will be lower when combining sorafenib with less intensive therapy such as 5-Azacytidine (5-Aza). This study was conducted to evaluate the efficacy and tolerability of the combination of sorafenib and 5-Aza in patients with refractory or relapsed AML with FLT3-ITD mutation. Methods: Patients were eligible if they had relapsed or refractory AML and were 18 years of age or older. They received 5-Aza 75 mg/m2 IV daily x 7 days and sorafenib 400 mg PO BID continuously; cycles were repeated in approximately one month intervals. Results: 38 patients with AML with a median age of 60 years (range, 24-87) were enrolled. 4 were inevaluable as they never received therapy or discontinued it before response assessment. FLT-3-ITD was detected in 30 (88%) patients. They had received a median of 2 prior treatments (range, 0-6); 13 (38%) had received ≥ 3 prior regimens and 8 had failed prior FLT3 kinase inhibitor. Response rate was 41%, including 6 (17%) with CRi, 4 (12) with CRp, 3 (9%) with CR, and 1(3%) PR. The median time to achieving CR/CRi was 2 cycles (Range, 1-6) and the median duration of CR/CRi was 3 months (Range, 1–12). Conclusions: Combination of 5-Aza and sorafenib is effective for patients with relapsed AML with FLT-3-ITD mutation.
Gene expression analysis of Notch signaling pathway receptors in colon cancer stem cells.
Authors: Papasotiriou Ioannis , Apostolou Panagiotis , Toloudi Maria , Chatziioannou Marina , Ioannou Eleni 
Published: February 2017

29 Background: The Notch signaling pathway is not only involved in cell differentiation, but also in carcinogenesis. Four different receptors are included in this pathway (Notch-1, Notch-2, Notch-3, and Notch-4) and each is encoded by a different gene. Recent data suggest that the activation of Notch-1 induces epithelial to mesenchymal transition (EMT) consistent with cancer stem cell (CSC) phenotype. It raises therefore the question of whether there is an interaction between the receptors. The present study aims to find out the gene expression patterns of Notch receptors in Colon CSCs, under the suppression of Notch-1. Methods: The experiments were performed in Colon CSCs provided by CELPOGEN. The stemness of these cells was tested both in gene and protein level with qPCR and flow cytometry assays for the stemness markers Oct4-CD44. The gene suppressed with RNAi assays and the knockdown was evaluated with molecular biology methods. The gene expression analysis was performed after RT-qPCR assays, by using the comparative Ct method (ΔΔCt) for the calculation of relative quantitation. Results: The Colon CSCs expressed Oct4-CD44 both in gene and protein level, indicating the stemness status. The knockdown of Notch-1 up to 75%, led to a reduction of gene expression in Notch-2 and Notch-3 receptors, while no significant difference was observed in Notch-4. The Notch-2 expression was decreased up to 90% while that of Notch-3 was at the same level with Notch-1. Conclusions: This research study attempts to find out any possible correlation between the Notch receptors in Colon CSCS. The experimental data indicate that Notch-1 interacts with Notch-2 and Notch-3 by affecting heavily their gene expression. It is therefore necessary to study if the other receptors are capable for EMT and then to find out which factors are involved. Further studies to a greater range of samples need to be performed, in order to use these data at clinical level.
Effect of targeted therapy on circulating tumor cells in patients with metastatic breast cancer.
Authors: Giordano Antonio , Barnato Sara E. , Giuliano Mario , Alvarez Ricardo H. , Cristofanilli Massimo , Hortobagyi Gabriel N. , Valero Vicente , Reuben James M. 
Published: February 2017

3 Background: Detection of ≥ 5 circulating tumor cells (CTCs) per 7.5 mL of blood before starting systemic treatment is a strong and independent predictor of worse progression-free survival (PFS) and overall survival (OS) in metastatic breast cancer (MBC) patients. It has been demonstrated that trastuzumab and bevacizumab can target CTCs in the peripheral blood of patients with breast cancer and modify the predictive value of the CTC count. We investigated time-related clinical trend of CTCs according to types of targeted therapy in MBC patients. Methods: Enumeration of CTCs was performed by the CellSearch technology as standard of care for 162 MBC patients receiving targeted therapy (trastuzumab, lapatinib, or bevacizumab) for MBC. The evaluation of CTCs was performed as follows: baseline CTC value defined as assay taken within 30 days before starting therapy, follow-up CTC value defined as the lowest CTC value after starting therapy. We analyzed the effect of treatments on CTC count by Wilcoxon matched pair test and estimated PFS and OS by Kaplan-Meier curves. Results: After a median follow-up of 26 months, 143 of 162 (88%) showed progression of disease and 96 (59%) had expired. Sixty-two (38%) patients have received treatment with trastuzumab, 31 (19%) patients with lapatinib, and 69 (43%) patients with bevacizumab. Among patients receiving anti-HER2 targeted treatments, trastuzumab and lapatinib, and those receiving bevacizumab, no differences in terms of median PFS and OS were observed between patients with CTCs greater or equal than 5 and less than 5. Analyzing separate anti-HER2 targeted treatments, we found trastuzumab decreased the number of CTCs at follow-up (P = 0.0003), whereas lapatinib did not (P = 0.14). However, bevacizumab also decreased the number of CTCs at follow-up (P < 0.0001). Conclusions: In MBC patients treated with targeted therapies, basal CTC count was not predictive of PFS and OS. Trastuzumab and bevacizumab significantly decreased the number of CTCs after starting treatment. Moreover, measuring CTCs during the follow-up of MBC patients may represent a predictive tool for targeted therapy failure.
Computed aided classification of circulating tumor cell and CK+ subclasses.
Authors: Gusyatin Oleg , Tims David , Milutinovic Aladin , Jiang Chunsheng 
Published: February 2017

54 Background: Fluorescence microscopy imaging system (OnQView, On-Q-ity, Waltham, MA) in combination with advanced cell capture techniques (OnQChip, On-Q-ity, Waltham, MA) provides necessary sensitivity to detect circulating tumor cells (CTCs) in a blood sample. The detection process involves automatic identification of CTC candidates from the collected imagery followed by CTC subclass identification. Subclass identification process is manual and usually leads to increased sample processing time. Methods: We have developed a fully automated CTC detection and classification system allowing for substantial increase in throughput while maintaining high sensitivity and specificity. Detection is accomplished by a robust segmentation technique. A set of 25 image-based features is automatically computed for each detected candidate. Features include texture measurements, morphology measurements, multichannel intensity and contextual characteristics. All CTC subclasses as well as artifact classes are manually labeled and verified by trained imaging technologists.A hierarchy of Multi-Layer Perceptron Neural Network (MLPNN) classifiers is then trained and used to identify and reject artifacts and to identify CTC subclasses. Results: A total of 27 prostate cancer patients and 33 normal controls with two 3.75ml blood samples per patient were used to validate techniques. Probability of successful artifact rejection was achieved to be 0.78 and probabilities of subsequent successful CTC subclass identification ranged between 0.79 and 0.98 (intact CTCs = 95%; irregular CTCs = 98%; fragmented CTCs = 82%). Conclusions: A fully automated CTC detection and classification system was developed. Testing was conducted with 27 prostate cancer patients and 33 normal controls to yield an artifact rejection probability of 0.78 and CTC subclass identification probabilities of 0.79 to 0.98.