bluuebunny/crossref_metadata_2025_split huggingface dataset:
Overcoming cell size escape from tumor treating fields using a varying frequency treatment paradigm in vitro.
Authors: Schneiderman Rosa S. , Giladi Moshe , Porat Yaara , Munster Mijal , Weinberg Uri , Kirson Eilon David , Palti Yoram 
Published: March 2020

e22134 Background: TTFields Therapy at 200 kHz received FDA approval for the treatment of patients with recurrent glioblastoma (GBM) based on the results of a phase III clinical trial. Radiological responses were observed in 14% of the treated patients. The lack of radiological response in the remaining patients suggests that GBM cells may escape the effect of TTFields over time. The goals of the present study were to identify possible routes by which cancer cells can escape the antimitotic effect of TTFields and to explore ways to overcome such escape mechanisms. Methods: Measurements of cell size before and after 72 hours TTFields application revealed a significant volume increase in 10 cancer cell lines (17% - 101%). A2780 ovarian cancer cultures (cell volume 2.0+0.2 pL) exhibit maximal inhibition by TTFields at 200 kHz. Twenty four hour application of 200 kHz TTFields led to a 70+29% increase in average cell volume (p<0.05). FACS analysis showed that TTFields application to A2780 cells led to a 44% increase in the G2 population due to the antimitotic effect of TTFields. However, the increase in the G2 proportion is not sufficient to explain the increase in cell volume seen after TTFields application. Furthermore, after an additional 24 hours of treatment, the fraction of cells in the G2 population returned to its original percentage (30%), while the increase in cell volume remained high (67+26%). Results: Previous publications demonstrated an inverse relationship exists between cell size and optimal TTFields frequency. Thus an increase in cell size may allow cells to avoid the maximal TTFields effect. To attempt to overcome such cell size escape we decreased the frequency of the TTFields from 200 to 150 kHz after 24 hours treatment to coincide with the increase in cell volume. We found that varying the frequency from 200 to 150 kHz increased the inhibitory effect of TTFields compared to continuous treatment at 200Khz (37+10% and 57+16% decrease in cell count, respectively; p=0.043). Conclusions: This is the first evidence of an escape mechanism from the antimitotic effect of TTFields. The results presented provide a simple way to restore treatment efficacy by varying TTFields frequency in parallel to the change in cell volume.
BIM deletion polymorphism and the clinical outcome of patients with advanced NSCLC who received first-line platinum-based chemotherapy: Preliminary results.
Authors: Zhang Jie , Zheng Di 
Published: March 2020

e22137 Background: The up-regulation of the proapoptotic polypeptide BIM was acquired during the cancer cell apoptosis induced either by cytotoxic agents or tyrosine kinase inhibitors (TKIs). Germline polymorphism of BIM deletion was associated with shorter progression free survival (PFS) in patients with EGFR-mutated lung cancer treated with EGFR-TKIs. The aim of this study is to investigate the correlation between the germline polymorphism of BIM deletion and the clinical outcome of patients with advanced NSCLC who received first-line platinum based chemotherapy. Methods: Since June of 2012, 140 consecutive chemo-naive advanced NSCLC patients who completed two cycles of first-line platinum based chemotherapy evaluable for response assessment were eligible for inclusion in Shanghai Pulmonary Hospital. Patients with stable disease were excluded in order to eliminate the indolent disease effect. Each patient’s genomic DNA was extracted from peripheral blood sample, then the genotype of the BIM deletion was performed using a single PCR reaction with the primers 5’-AATACCACAGAGGCCCACAG-3’ and 5’-GCCTGAAGGTGCTGAGAAAG-3’. Results: There are 49 patients who responded to chemotherapy (RE group) while 32 patients developed progressive disease (PD group). The clinical characteristics at baseline and chemotherapy regimens received were balanced between the two groups except for more stage IV patients noted in PD group (57 vs 58 years old of median age, 59% vs 62.5% of male, 63% vs 100% of stage IV in RE group and PD group respectively, and the predominant regimen with gemcitabine plus cisplatin in both groups). BIM deletion polymorphism were detected in 4 of RE group (4/49, 8.2%) and 8 of PD group (8/32, 25%) respectively, (odds ratio=0.267, 95% CI=0.073-0.977, P=0.046). Conclusions: The germline polymorphism of BIM deletion may contribute to the intrinsic resistance to the chemotherapy in advanced NSCLC.
Nuclear localization of the androgen receptor in prostate cancer circulating tumor cells from men who have failed androgen deprivation therapy.
Authors: Lang Joshua Michael , Casavant Benjamin Pun , Berry Scott Michael , Strotman Lindsay Nicole , Thiede Stephanie Margaret , Sperger Jamie , Liu Glenn , Beebe David J. 
Published: March 2020

e22141 Background: Prostate cancer (PRCA) is driven by the Androgen Receptor (AR), a cytoplasmic protein that translocates to the nucleus to activate a transcriptional program critical to PRCA tumorigenesis. The AR is the major therapeutic target in PRCA, with multiple drugs that improve survival and quality of life for men with advanced PRCA. However, few biomarkers can assay for AR activity to identify those most likely to benefit from AR-targeting therapies. Methods: Utilizing a novel microfluidic technology known as the VERSA (Vertical Exclusion-based Rare Sample Analysis) platform, CTCs were captured and purified from men with advanced PRCA who had failed androgen deprivation therapy. CTCs were then stained and imaged via epiflourescent microscopy for EpCAM, AR, Hoechst and CD45. The enhanced optical imaging of the VERSA platform enabled subcellular localization of the AR and calculation of the relative amount of AR localized to the nucleus as a biomarker of resistance to AR-targeting therapies. Results: CTCs were prospectively collected and analyzed for AR nuclear localization. Significant heterogeneity in AR localization was found between patients who had failed androgen deprivation therapy with the mean nuclear AR ranging from 20-57% of total AR. Significant intra-patient heterogeneity was also identified with the percentage of AR localized to the nucleus ranging from 11-77% of total AR. DNA and mRNA have been isolated from these cells to analyze for underlying mechanisms of nuclear localization. Conclusions: The VERSA platform is able to analyze subcellular localization of proteins in CTCs followed by total nucleic acid extraction. We show that among men whose disease has progressed through androgen deprivation, a significant portion of the AR is localized to the nucleus as a possible biomarker of resistance to AR targeting therapies. We also identify significant heterogeneity in AR localization among CTCs, suggesting resistant PRCA clones that have greater proliferative potential.
Long-term stability of RNA from FFPE brian tissue post-sectioning on slides.
Authors: Witt Donald James , Anderson Steven M. , King Briana , Borrego Christina , Eisenberg Marcia , Cai Li 
Published: March 2020

e22142 Background: Analysis of nucleic acids (NA) from formalin fixed paraffin embedded (FFPE) tissue can provide detailed information about gene sequence mutational status, which may be important for oncology treatment decisions. FFPE specimens also have utility for retrospective analyses. Potential degradation of NA during formalin fixation, paraffin embedding processes and possible continued deterioration during subsequent storage may diminish utility of FFPE tissue for these purposes. Using real-time PCR, this study investigated the functional stability of RNA from brain FFPE tissue sections on slides over an extended time period after sectioning. Methods: Brain biopsy specimens obtained from glioblastoma patients with informed consent were used to prepare blocks with standard formalin fixation and paraffin embedding techniques. Slides were made from the FFPE blocks and stored at room temperature until testing. RNA was extracted from sequential slides within one week of sectioning for a zero time and then at 4, 8 and 12 months. Reverse transcription PCR was performed, and real-time PCR was analyzed on the ABI7900 to detect EGFRvIII mutation and cABL gene. RNA Integrity Analysis was performed with an Agilent Bioanalyzer. Results: Consistent qualitative results were obtained with EGFRvIII mutant positive specimens (n =10) and wild type (wt) specimens (n =10) from slides stored up to twelve months at room temperature compared to the initial testing (95% agreement). One wt specimen showed negative results for the first three time points but a low positive result at 12 months, possibly due to tumor content change in the different sections of the FFPE block. Ct values for EGFR (wt and mutant) and cABL genes did not increase during the storage period. RNA integrity number (RIN) indicated the degradation of RNA during FFPE processing, although no further significant degradation occurred during the course of the experiment. Conclusions: The results of this study indicated that although the RNA was impacted by the tissue preparation, fixation, and processing steps, for the brain FFPE slide specimens, target genes with amplicon size up to 124bp could be detected with minimum degradation for up to 12 months when slides were stored at room temperature.
Annexin A1 and HPV in oropharyngeal squamous cell carcinoma.
Authors: Queiroz Cleberson Jean dos Santos , Nakata Cintia Mara de Amorim Gomes , Damazo Amilcar Sabino 
Published: March 2020

e22150 Background: Annexin-A1 (ANXA1) is a protein involved in signal transduction and inflammation. Its expression is increased in some tumors (i.e. hepatocellular carcinomas, gliomas, pancreatic adenocarcinomas), but in head and neck tumors, its expression is usually reduced. Tumor suppressor p53 is frequently expressed in cancers due to mutation. HPV infection is strongly linked to oropharyngeal tumors. Methods: We analyzed 21 formalin-fixed paraffin-embedded samples of oropharyngeal carcinoma. Using immunofluorescence (IF), ANXA1 expression was quantified by measuring mean optical density (MOD). We also evaluated p53 positivity using IF. Presence of HPV DNA (serotypes 16/18 and 31/33) was analyzed through chromogenic in situ hybridization. Interaction between ANXA1, p53 and HPV was performed by Student's t test and ANOVA, with Bonferroni test, using the software Graph Pad Prism. Results: HPV DNA was found in 12 of 21 cases (57%). Serotypes 16/18 were found in all HPV + samples, whereas serotypes 31/33 were found in 2 of the 12. Among HPV+ cases, we noticed a decreased expression of ANXA1 in tumor compared to epithelium, independent of p53 status. With respect to HPV- cases, we have found a reduced expression of ANXA1 in the tumor versus epithelium, but the difference was only statistically significant in p53+ samples. In epithelia, we observed an increased expression of ANXA1 in HPV+ compared with HPV- samples. Conclusions: Our results confirm a decreased expression of ANXA1 in oropharyngeal carcinoma independently of HPV status, suggesting its involvement in the carcinogenesis. There was no difference in the expression of ANXA1 according to p53 status. In normal epithelia, we observed an increased expression of ANXA1 in HPV+ samples, which may suggest the involvement of the protein in the early stages of HPV-driven carcinogenesis. [Table: see text]
Characterization of DNA methylation of the promoters CTCF and BORIS (CTCFL) in breast and ovarian cancer.
Authors: Soto Reyes Solis Ernesto , Morales-Espinosa Daniela , Cantu David , Alvarado-luna Gabriela , Green Dan , Castro Hernandez Clementina , Alcala-Moreno Iliana , Cisneros-Soberanis Fernanda , Motola Daniel , Rodriguez-Cid Jeronimo , Herrera Luis A. 
Published: March 2020

e22151 Background: Genetic and epigenetic alterations may promote the initiation or development of cancer. Global DNA hypomethylation and local hypermethylation have been observed, particularly in cell cycle control-associated genes, such as tumor suppressor genes like CTCF. The dissociation of CTCF is associated with hypermethylation of several promoters; its paralogue gene (BORIS) is normally expressed in testicular tissue during spermatogenesis. BORIS over-expression has been identified in multiple neoplasms such as melanoma, gynecological cancer, glioblastoma and – recently – breast cancer. The aim of this study was to characterize the methylation status of the promoter regions of CTCF and BORIS in samples from breast and ovarian cancer compared to non-neoplastic tissue, and correlate it to its expression. Methods: Tissue samples from breast and ovarian cancer, as well as healthy controls were analyzed by MS-PCR for CTCF and BORIS. BorismRNA expression was also analyzed by RT-PCR. Results: A total of 8 ovarian and 16 breast tumors, as well as 10 tumor-adjacent breast tissue samples were prospectively obtained. In non-neoplastic tissue, BORIS was found to be hypermethylated, while in ovarian tumors a loss of methylation was identified in 75% of the samples. The same phenomenon was observed in 68% of breast cancer samples when compared to non-neoplastic tissue. A correlation between loss of DNA methylation of the promoter and gene over-expression was found by RT-PCR, thus suggesting that methylation is an epigenetic phenomenon associated to the over-expression of the oncogene BORIS. The methylation analysis of CTCF did not show any differences between neoplastic and non-neoplastic tissue, suggesting that epigenetic changes mainly affect BORIS. Conclusions: Loss of methylation of the promoter region of BORIS is associated with the over-expression of the gene. No differences were found in the methylation status between healthy and neoplastic tissue for CTCF.
Relative importance of oncologist access and patient pre-selection in limiting the uptake of companion diagnostic tests for solid cancers in the United States.
Authors: Ilacqua Jackie , De Richter Pieter ,   Ipsos Healthcare, Inc.
Published: March 2020

e22157 Background: The speed of uptake of Companion Diagnostics (CDx) tests in the oncologist community is a product of (1) the number of physicians adopting the test in the months following launch, and (2) the % of patients under those physicians’ care who are being tested. Here, we investigate which one of the two factors was the most significant in terms of limiting the speed of uptake within the initial 3 months after launch of two recently approved CDx tests in the U.S. Methods: A panel of oncologists in the US (N=130) were asked to report on their stated tested levels of EML4-ALK testing for their current stage IIIB and IV NSCLC patients, and BRAF mutation testing for their current stage IV melanoma patients. This data was collected through an online survey between August and November 2012. Results: Of the 130 physicians surveyed, only 5 (4%) did not use EML4-ALK testing for their patients at all, and just under 33% used EML4-ALK testing for all their stage IIIB/IV NSCLC patients. The majority (64%) used EML4-ALK testing for a subset of their stage IIIB/IV NSCLC patients only, indicating they preselected their patients for testing. This compares to as many as 55% of surveyed physicians using BRAF for 100% of their stage IV melanoma patients, with a further 29% using it for a subset of their stage IV melanoma patients and 7% not using the test at all. Conclusions: As the vast majority (96% and 93% respectively) of surveyed physicians used EML4-ALK and BRAF testing for at least a subset of their patients within the first 6 months after launch, a lack of access to the tests did not appear to be a major barrier to uptake. Instead, the % of an individual oncologist’s patients being tested through pre-selection for testing was the primary factor determining the volume of testing. Pre-selection was more common for EML4-ALK testing than for BRAF testing, possibly due to the higher incidence of BRAF mutations than EML4-ALK translocations.
Dynamic real-time imaging of marrow-resident neutrophils interacting with human mesenchymal stem cells during systemic lipopolysaccharide challenge.
Authors: Huang Alex Yee-Chen , Myers Jay T. , Barkauskas Deborah Sim 
Published: March 2020

e22158 Background: Human mesenchymal stem cells (hMSCs) have gained intense interest due to their immune-modulatory, tissue differentiating and homing properties to sites of inflammation and tumor microenvironment, as evidenced by more than 200 ongoing clinical trials using hMSCs in a variety of clinical settings. Despite evidence demonstrating the bio-distribution of infused hMSCs in target organs using static fluorescence imaging or whole-body imaging techniques, there is controversy regarding how hMSCs exert their biological effects, and very little is known about how they behave dynamically within host tissues on a single-cell level in vivo. Methods: We infused fluorescently labeled clinical-grade hMSCs into immune-competent mice in which neutrophils and monocytes express a second fluorescent marker under the Lysozyme M (LysM) promoter. The recipient mice were then subjected to serial 4-D (xyzt) imaging of the bone marrow cavity with intravital two-photon microscopy (TPM) during acute systemic Lipopolysaccharide (LPS) challenges to observe changes in MSC and neutrophil migration behavior. Results: We were able, for the first time, to capture dynamic interactions between and migration pattern of hMSCs and LysM+granulocytes in the bone marrow of live mice during systemic LPS challenge. Contrary to some published reports, many of the infused hMSCs remained intact despite repeated cellular contacts with host neutrophils. However, we also observed the destruction and subsequent phagocytosis of some hMSCs by surrounding granulocytes. Conclusions: Our imaging platform provides opportunities to gain insight into the biology and therapeutic mechanisms of hMSCs in vivo at a single-cell level within live hosts.
The clinical significance of ephrin receptors expression in non-small cell lung cancers.
Authors: Tsoukalas Nikolaos , Giaginis Constantinos , Kavantzas Nicolaos , Alexandrou Paraskevi , Bournakis Evangelos , Patsouris Efstratios , Theocharis Stamatios 
Published: March 2020

e22160 Background: Ephrin receptors (Ephs) are frequently overexpressed in a wide variety of human malignant tumors, being associated with tumor growth, invasion, metastasis and angiogenesis. The present study aimed toevaluate the clinical significance of Eph-A1, -A4, -A5 and -A7 protein expression in non-small cell lung cancers (NSCLC). Methods: Eph-A1, -A4, -A5 and -A7 protein expression was assessed immunohistochemically in tissue microarrays of 88 surgically resected NSCLC cases and was analyzed in relation with clinicopathological characteristics and patients’ survival. Results: Elevated Eph-A4 expression was significantly associated with earlier or lower stages and histopathological presence of inflammation (p=0.047 and p=0.026, respectively). Elevated Eph-A7 expression was significantly associated with patients’ age, presence of fibrosis and smaller tumor size (p=0.036, p=0.029 and p=0.018, respectively). NSCLC patients with elevatedEph-A4, -A5 or -A7 expression presented significantly longer overall survival times compared to those with reduced Eph-A4, -A5 or -A7 expression (log-rank test, 0=0.019, p=0.006 and p=0.012, respectively). Eph-A4, -A5 and -A7 expression were identified as independent prognostic factors of patients’ survival after adjustment for histopathological type and stage and performance status (Cox-regression analysis, p=0.022, p=0.033 and p=0.037, respectively). Conclusions: The present study supported evidence that Ephs may play a role in lung cancer progression, reinforcing their utility as clinical biomarkers for patients’ management and prognosis, as well as targets for potential therapeutic intervention in the future.
Isolation of a novel IgG from a patient with lung cancer using immunoassays and cloning.
Authors: Dessain Scott K. , Borghaei Hossein , Zhang Jiping , Simon Paul , Huang Minzhou , Zhao Huiwu , Puligedda Rama Devudu , Murphy Maureen E. 
Published: March 2020

e22163 Background: Cancer cells express novel antigens that are characteristic of the transformed state and may engender an immune response ( i.e., antibody production) in patients. These antibodies may identify novel immune targets for antibody-based immunotherapies and diagnostics. Furthermore, the presence of these auto-antibodies could be followed as biomarkers of therapeutic response or relapse. Previous studies of the antibody response to tumors have been hampered by inefficient methods of cloning the human antibodies in their native configurations. We collected blood samples from a group of lung cancer patients with diverse clinical and pathologic features and are characterizing the anti-tumor antibody response using a variety of whole-cell binding assays. Our approach also uses an optimized hybridoma method to clone tumor-specific IgG antibodies from primary memory B-cells. Methods: We obtained serum and peripheral blood mononuclear cell samples from 50 patients with advanced lung cancer. We characterized the immune response of these patients to tumor cell lines using fluorescence microscopy with the Operetta (Perkin Elmer) and by whole-cell ELISA. We performed a cell fusion using B-cells from one of the immunoreactive patients, screening the hybridomas for antibodies specific for binding in the whole cell ELISA. We characterized the antibody activity using flow cytometry and immunoblotting, and we identified the antigen using mass spectroscopy analysis of a gel-purified, immunoreactive band. We confirmed the identity of the antigen bound by immunoblotting to recombinant protein. Results: Human IgG antibodies immunoreactive with human cancer cell lines are prevalent in our panel of lung cancer patients. Using our hybridoma method, we isolated a human IgG that binds a tumor-specific surface antigen expressed in a variety of tumors, but only minimally expressed by normal cells. Conclusions: Novel IgG antibodies against specific tumor antigens can be isolated from lung cancer patients’ serum.Using our novel hybridoma method for cloning human antibodies, we have identified an antibody that binds a tumor-specific antigen. The study of these antibodies may lead to novel biomarkers for cancer diagnosis and therapy.