bluuebunny/crossref_metadata_2025 huggingface dataset:
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.
62cuatsm and 62cuptsm PET is a useful imaging tool for hypoxia and perfusion in lung cancer.
Authors: Zhang Tian , Das Shiva K. , Crawford Jeffrey , Wong Terence , Dewhirst Mark W. , Vlahovic Gordana
Published: March 2020

e22166

Background: Hypoxia is a characteristic of many tumors and portends a worse prognosis in lung, cervical, prostate, and rectal cancers. Unlike the others, lung cancers present a unique challenge in measuring hypoxia, with invasive biopsies and higher rates of complications. Noninvasive imaging studies detecting hypoxia using isotopes of copper-diacetyl-bis (N4-methylthiosemicarbazone), CuATSM, have predicted prognosis and treatment outcomes in some small feasibility trials. These images, however, may not identify all areas of hypoxia. Hence, we hypothesize that the addition of another positron emission tomography (PET) imaging agent, copper-pyruvaldehyde-bis (N4-methylthiosemicarbazone), 62CuPTSM, which can detect areas of perfusion, can augment the information obtained in 62CuATSM PET scans. Methods: In order to characterize tumors based on both perfusion and hypoxia, ten patients were studied using both 62CuATSM and 62CuPTSM PET scans. All patients signed informed consent; the protocol was approved by the Duke University Medical Center institutional review board. 62CuATSM and 62CuPTSM SUV values at steady state were tabulated and the ratios of 62CuATSM to 62CuPTSM calculated. In addition, proteomic arrays looking at specific proangiogenic, survival, and proinflammatory targets were assessed. Results: Six out of ten patients had fully evaluable PET scans. Our initial experience of characterizing lung tumor hypoxia using 62CuATSM/62CuPTSM PET scans showed that visualization of areas with hypoxia normalized for perfusion is feasible. All studied tumors exhibited some hypoxia. Despite the small sample size, a positive relationship was noted between serum epidermal growth factor (EGF) levels and 62CuATSM detected hypoxia. Conclusions: This initial series of 62CuATSM/62CuPTSM PET scans demonstrate that evaluating pulmonary lesions by visualizing hypoxia and perfusion is a feasible and novel technique to provide more information. Further investigation is warranted to assess the potential role of 62CuATSM and 62CuPTSM PET imaging techniques combined with proteomics as alternatives to invasive biopsy techniques in clinical care.
Time for early predictive biosignatures to personalize the clinical approach of advanced tumor patients?
Authors: Yamaguchi Nise Hitomi , Spada Manoela Tavares Arruda , Ribeiro Rachel , Silverio Flavio , Tamura Paula , Moura Fernando , Oliveira Aline Crisitna Conceicao , Cordeiro Eivete , Anciaes Fabiana Sequim , Kobayashi Kleber , Donatangelo Silvio , Oliveira Nivia Fernanda
Published: March 2020

e22176

Background: The Institute of Advances in Medicine is a private referral center in Brazil that treats patients with advanced disease, often after failure to previous lines of treatments. Predictive response assays allow physicians to make better informed choices when establishing the therapeutic strategy for each patient. Methods: 48 patients (21 male; 27 female), 54.5 y old (33-80y) with advanced disease were submitted to molecular diagnostic investigation using paraffin-embedded tumor samples from Brazil, shipped to Caris Target Now-US in accordance with the Brazilian Federal Sanitary Agency regulations. We have considered the samples diversity, different tumor sites, tissue availability and technical conservation. Results: More than 1920 IHC, FISH or CISH, RT-PCR, RFLP, sequencing and mutational analysis by PCR were performed, with a list of drugs with Potential Clinical Benefit (PCB) or Potential Lack of Benefit (PLB) for the 48 primary tumors: 11 colorectal, 9 breast, 6 NSCLC, 22 other sites. Drugs were listed in the PCB column that were usual drugs in first and second line treatments for their cancer types. However, many patients were classified with PLB with drugs that they had used before and were not active. These data were analyzed for Performance Status, Previous Treatments actually given and current commercially available anticancer agents, such as chemotherapeutic and new molecular targeted agents to help the decision for future treatments. Conclusions: When the test was performed, patients had already been treated (and had had disease progression) with drugs that were listed as potentially lacking benefit. Time is ripen for testing tumors prior to beginning clinical treatment, since it may provide a better understanding of the tumor biology and the probable clinical response, making it possible to distinguish which therapeutic schedule is preferable. The present availability of reliable predictive assays for assessing tumor sensitiveness (or lack of it) to anticancer drugs makes it imperative to change guidelines. The ever growing health costs in the public health systems around the world may also benefit from such strategy since only drugs with a potential for response will be chosen.
A novel approach to improve detection of somatic DNA variants in solid tumors by next-generation sequencing from FFPE samples.
Authors: Udar Nitin , Haigis Robert , Gros Thomas , Lucio-Eterovic Agda Karina , Zhang Qingxiu , Zenali Maryam , Ross Mark , Jaeger Erich
Published: March 2020

e22177

Background: Low frequency variant detection by sequencing is a highly desired goal for therapy selection in cancer especially the detection of actionable targets. The lower limit of detection using Sanger sequening is ~20% minor allele frequency (MAF). Deep sequencing of target genes using next generation sequencing (NGS) is gaining popularity. Formalin Fixed Paraffin Embedded (FFPE) tissue is the most common sample type in solid tumor histopathology. However, because the fixation process fragments DNA and damages it at varying frequencies, downstream processes can potentially misclassify modified bases and generate artifacts. We have developed a protocol that addresses both of these issues in a multiplex assay that involves deep sequencing using NGS of targets implicated in lung, gastric, colon, melanoma and ovarian cancers. Methods: The sample set includes 168 FFPE samples and 82 non FFPE samples. TruSeq Custom Amplicon technology was used to generate libraries for targets across 26 genes. Deep sequencing was done on a NGS platform (Illumina-MiSeq). Results: The DNA quality test, which surveys multiple genomic targets by qPCR, was an accurate determinant of DNA amplifiability and yielded a 99% sample success rate. A sensitivity of <5% MAF was achieved by sequencing at a depth of 1,000X for all targets. In order to differentiate true low frequency variants from fixation and other artifacts, our novel approach investigates each of the two DNA strands independently. The information is bioinformatically combined to distinguish true variants from artifacts. Testing of the FFPE samples with a 5% MAF cut off using the two strand approach reduced the potential false positive rate by ~ 40% when compared to information from only one strand of DNA. A comparative analysis of matched FF and FFPE sample showed that a high percentage of false positive calls were present even in the fresh frozen samples at this high level of sensitivity, if only using information from one strand. Conclusions: This protocol, efficiently and accurately detect low frequency variants by NGS in DNA extracted from FFPE tissues.
Development of an automated RNA-based nuclease protection assay for the detection of expressed EML-ALK fusions, EGFR, and KRAS mutations in NSCLC.
Authors: Rounseville Matthew , Maddula Krishna , Seligmann Bruce , Botros Ihab , Modur Vijay
Published: March 2020

e22185

Background: The management of patients for NSCLC requires appropriate molecular pathology assessment to determine treatment. However current methods require the use of more than one technology including sequencing for EGFR and KRAS mutation and in situ hybridization for EML-ALK fusion, resulting in slow turnaround, and technical as well as logistical complexities. Here, we report the development of an automated microarray based quantitative nuclease protection assay that determines gene fusions, deletions and point mutations in target genes such as EML4/ALK, EGFR and KRAS from expressed mRNA directly from FFPE tissue without prior RNA extraction. Methods: We designed mutation and fusion specific probes for nuclease protection and measured the sensitivity, specificity, and reproducibility of gene mutations and fusions using in vitro transcribed RNA and cell lysates known to contain the target mutations. In addition to synthetic samples, we also screened 88 lung cancer specimen for EML-ALK fusions. Results: Mutations in KRAS (G12D, G12C, G12V); EGFR (D761Y, T790M, L858R); EML-ALK fusions (v1 v2, v3a, v3b-3, v4, v5a, v5b-3 and v6) were detected at concentrations of 3 copies/cell in 15,000 cells even when present as low as 5% of wild type IVT RNA. The sensitivity corresponds to an amount of FFPE sample ranging from 0.3 to 0.6 square cm of a 5mm FFPE section per replicate assay making it suitable for needle biopsy specimen. Six samples with probable EML4-ALK fusions were identified in the 88 adenomatous NSCLC samples and will be subjected to further confirmation. Additional clinical validation of all the assays will be presented in our poster. Conclusions: An automated, multiplexed, EGFR and KRAS mutation and EML-ALK gene fusion from expressed mRNA is feasible. The assay can be run directly on FFPE sections, does not require RNA extraction or amplification, and is completed within 24 hours.
Aneuploidy and neoplasia: Case of recurrent polysomy occurring at different times.
Authors: Adiga Giridhar U. , Ramesh K H , Tong Tommy R , Jodlowski Tomasz
Published: March 2020

e22196

Background: Recurring polysomies in diverse tumors at different times may suggest common neoplastic mechanisms. Following case demonstrates similar polysomies in breast, endometrial and cholangiocarcinoma arising at different times. Methods: A 79-year-old African American female was diagnosed poorly differentiated cholangiocarcinoma. Tumor markers (CEA, AFP, CA19-9) were normal. FISH analysis with D3Z1 (centromere 3), D7Z1 (centromere 7), CDKN2A (9p21) and D17Z1 (centromere 17) DNA probes (Abbott Molecular, IL. USA) revealed polysomy of chromosomes 3, 7, 9 and 17. Oncology history includes tumors noted in the table. Medical history includes aortic aneurysm, hypertension, IPF, obesity, CKD, DJD, cholecystectomy and adjuvant tamoxifen. She was never pregnant, non smoker non alcoholic and no prior chemical exposures. Her maternal aunt had unclear pelvic cancer at 70. Her niece had duodenal cancer in 40s. Possible common initiating neoplastic mechanism or predisposition was speculated in view of multiple tumors. Archived specimens of previous tumors were tested using the same FISH probe set. Results: Similar pattern of aneuploidy was noted in cholandiocarcinoma, DCIS and endometrial polyp specimen and no such abnormalities in parathyroid adenoma and in normal tissue. Conclusions: Occurrences of similar polysomy in tumors originating from diverse tissues at different times but not in normal tissue suggest common predisposition and/or similar, nonrandom, neoplastic molecular events. Uncommon and previously not described polysomy pattern was noted in DCIS and in endometrial polyp. [Table: see text]
Clinical significance of cytokines levels in exhaled breath condensate (EBC) and serum of lung cancer patients.
Authors: Lamprodimou Georgia , Athanasiadis Athanasios , Kerenidi Theodora , Doufexis Dimitrios , Lada Martha , Tsilioni Irini , Roussos George , Vavougios George , Gourgoulianis Konstantinos
Published: March 2020

e22202

Background: Cytokines are known to play an important role in carcinogenesis.The aim of this study is to measure pre-treatment concentrations of cytokines IL6, IL10, IL1β and IL8, in EBC and serum of lung cancer patients and to assess any possible association with clinicopathological variables and survival. Methods: From February 2010 to May 2011 we enrolled 73 patients with previously untreated lung cancer, 67 males and 6 females, age 67.7±9.5 years, 46 smokers and 27 ex-smokers, and 30 healthy individuals as controls. Sixty-one patients had NSCLC and 12 SCLC (5 stage I, 3 stage II, 19 stage III and 46 stage IV). EBC was collected using the Ecoscreen device (Viasys, Germany). Serum and EBC levels of cytokines were analyzed by commercially available enzyme-immunosorbent assay kits (ELISA). Results: All cytokines but IL8 were detectable in both materials. IL8 was not detected in EBC. All cytokines were significantly increased in patients with lung cancer in comparison to controls (IL6: serum P<0.0001, EBC P=0.001; IL10: serum P= 0.006, EBC P=0.037; IL1β: serum P= 0.032, EBC P=0.005; IL8 serum P=0.001). Moreover, a statistically significant correlation was found between IL10 levels in EBC and serum (P=0.001, Rho=0.334). In EBC, higher levels of IL6 were found in patients with T3-T4 disease than in those with T1-T2 [4.27 pg/ml (2.46-5.99) vs 3.12 pg/ml (1.35-5.03), P=0.036]. Furthermore, smokers had higher serum levels of IL6 compared to ex-smokers [10.19 pg/ml (4.92-17.38) vs 4.46 pg/ml (1.88-11.53), P=0.009]. Finally, lower serum levels of IL6 < median value (9.55pg/ml) were associated with improved survival (median overall survival 338 days vs 237 days Log Rank test, P= 0.046). Conclusions: Elevated levels of measured cytokines were found in both EBC and serum of lung cancer patients. Particularly, IL6 levels were related to T-stage and smoking status, while lower serum levels of IL6 with survival. These findings suggest that cytokines should be further studied in order to evaluate their diagnostic and prognostic significance. EBC could provide a simple, noninvasive and economic method for the monitoring of biomarkers in patients with lung cancer.
Lapatinib in combination with capecitabine plus oxaliplatin (CapeOx) in HER2 positive advanced or metastatic gastric (A/MGC), esophageal (EAC), or gastroesophageal (GEJ) adenocarcinoma: The LOGiC trial.
Authors: Hecht J. Randolph , Bang Yung-Jue , Qin Shukui , Chung Hyun-Chul , Xu Jian-Ming , Park Joon Oh , Jeziorski Krzysztof , Shparyk Yaroslav , Hoff Paulo M. , Sobrero Alberto F. , Salman Pamela , Li Jin , Protsenko Svetlana , Buyse Marc E. , Afenjar Karen , Kaneko Tomomi , Kemner Allison , Santillana Sergio , Press Michael F. , Slamon Dennis J.
Published: March 2020

LBA4001

The full, final text of this abstract will be available at abstract.asco.org at 7:30 AM (EDT) on Monday, June, 3, 2013, and in the Annual Meeting Proceedings online supplement to the June 20, 2013, issue of Journal of Clinical Oncology. Onsite at the Meeting, this abstract will be printed in the Monday edition of ASCO Daily News.
Correlation of molecular alterations with efficacy of everolimus in hormone-receptor–positive (HR+), HER2-negative advanced breast cancer: Preliminary results from BOLERO-2.
Authors: Hortobagyi Gabriel N. , Piccart-Gebhart Martine J. , Rugo Hope S. , Burris Howard A. , Campone Mario , Noguchi Shinzaburo , Perez Alejandra T. , Deleu Ines , Shtivelband Mikhail , Provencher Louise , Masuda Norikazu , Dakhil Shaker R. , Anderson Ian , Chen David , Damask Amy , Huang Alan , McDonald Robert , Taran Tanya , Sahmoud Tarek , Baselga José
Published: March 2020

LBA509

The full, final text of this abstract will be available at abstract.asco.org at 7:30 AM (EDT) on Monday, June, 3, 2013, and in the Annual Meeting Proceedings online supplement to the June 20, 2013, issue of Journal of Clinical Oncology. Onsite at the Meeting, this abstract will be printed in the Monday edition of ASCO Daily News.
Trial of zoledronic acid and interleukin-2 to expand tumoricidal γδ T cells in vivo in patients with refractory neuroblastoma.
Authors: Pressey Joseph Gerald , Adams Julia M , Kelly David R , Lamb Lawrence S
Published: March 2020

TPS10073

Background: CD3+ γδ+ T cells comprise 2 to 5% of the circulating T cells. Vγ9Vδ2+ cells are the dominant circulating γδ T cell and recognize non-peptide phosphoantigens and stress-associated NKG2D ligands expressed on malignant cells. Strategies that incorporate the tumoricidal properties of γδ T cells represent a promising means of harnessing the innate immune system to treat malignancies including neuroblastoma (NB). Indeed, γδ T cells from both healthy volunteers and NB patients exert a potent cytotoxic effect on NB cell lines and autologous NB in vitro following expansion and activation in culture. Vγ9Vδ2+ cells can also be induced to proliferate in vivo. By blocking farnesyl pyrophosphate synthase in the mevalonate pathway of isoprenoid synthesis in monocytes, the aminobisphosphonate zoledronic acid (ZOL) promotes the accumulation of isopentenyl pyrophosphate which is sensed by γδ T cells. IL-2 is also required for robust expansion γδ T cells. Methods: The trial is a prospective, non-randomized trial that assesses two dose levels of recombinant IL-2 (aldesleukin) in combination with ZOL. To be eligible for the study patients must be 2 to 21 years of age with refractory neuroblastoma with no known curative therapeutic options. Patients must also have adequate organ function and performance status. ZOL is given intravenously on day 1, and aldesleukin is given subcutaneously on days 1 to 5 and 15 to 19 of every 28 day cycle. The single-institution study is being conducted at the University of Alabama at Birmingham. Correlative studies include evaluating the absolute count, phenotype, activation, and effector/memory progression of γδ T cells by flow cytometry and the biological function of autologous expanded/activated γδ T cells by in vitro cytotoxicity assays employing established human NB cell lines. In order to determine the ability of in vivo expanded/activated γδ T cells to infiltrate NB tissue, bone marrow core biopsies obtained before and after the first course of therapy from patients with bone marrow metastasis are assayed for T cell infiltration by immunohistochemistry. The study has currently enrolled 1 of the planned 6 patients. Clinical trial information: NCT01404702.
A randomized, double-blinded, placebo-controlled, multi-institutional, cross-over, phase II.5 study of saracatinib (AZD0530), a selective Src kinase inhibitor, in patients with recurrent osteosarcoma localized to the lung.
Authors: Baird Kristin , Reinke Denise K. , Pressey Joseph Gerald , Mascarenhas Leo , Federman Noah , Marina Neyssa , Chawla Sant P. , Lagmay Joanne Pigues , Goldberg John M. , Milhem Mohammed M. , Loeb David Mark , Butrynski James E. , Janeway Katherine A. , Turpin Brian , Staddon Arthur P. , Spunt Sheri L. , Rodler Eve T. , Schuetze Scott , Okuno Scott H. , Helman Lee J.
Published: March 2020

TPS10591

Background: Osteosarcoma is a rare cancer and 33% of patients who have completed primary treatment will recur. The Src pathway has been implicated in the metastatic behavior of several tumors including osteosarcoma where 95% of samples express Src or have evidence of downstream activation of this pathway. Saracatinib (AZD0530) is a potent and selective Src kinase inhibitor. The recommended phase II dose in adults was found to be 175mg daily. The primary goal of this study is to determine if treatment with Saracatinib can increase progression free survival (PFS) for patients who have undergone complete resection of metastatic osteosarcoma nodules in the lung. Secondary goals are evaluation of overall survival, time to treatment failure, and evaluation of several biological correlatives. Methods: This is a multi-institutional, phase II.5, placebo-controlled study with an accrual goals of 88 randomized patients. Patients between 15 and 75 years, with histological confirmation of recurrent osteosarcoma, localized to the lung, who have potential for complete surgical resection, are eligible for enrollment. After complete resection, patients are randomized to treatment with saracatinib or placebo, of a daily oral dose of 175 mg, continuously for up to 1 year or until progression. Patients who recur in the lung while on-study and who are amenable to complete surgical resection will be un-blinded. Those patients who received placebo may have the option to undergo surgical resection. If fully resected, they will be offered therapy with saracatinib under the same treatment guidelines as above. As of January 2013, 38 patients have enrolled and 32 patients met the criteria to be randomized and began oral therapy with either saracatinib or placebo. An interim analysis is planned after 40 patients have been randomized. Clinical trial information: NCT00752206.
The prospective MammaPrint MINT (Multi-Institutional Neo-adjuvant Therapy) study.
Authors: Cox Charles E. , Blumencranz Peter William , Saez Ruben A. , Wesolowski Robert , Stork Lisette , De Snoo Femke , Gibson Jessica , Howard Nicole , Avisar Eli
Published: March 2020

TPS11122

Background: Patients with locally advanced breast cancer (LABC) are often treated with neo-adjuvant chemotherapy to reduce the size of the tumor before definitive surgery. Complete pathologic Response (pCR) predicts better long term outcome. Genomics assays that measure specific gene expression patterns in a patient's primary tumor have become important prognostic and predictive tools for early breast cancer. This study is designed to test the ability of molecular profiling, as well as traditional pathologic and clinical prognostic factors to predict responsiveness to neo-adjuvant chemotherapy in patients with LABC. Methods: Women ≥ 18 yrs with histologically-proven invasive breast cancer T2(≥3.5cm)-T4,N0M0 or T2-T4N1M0, with measurable disease, adequate bone marrow reserves and normal renal and hepatic function who signed informed consent are enrolled. Axillary lymph nodes will be staged according to protocol. MammaPrint risk profile, BluePrint molecular subtyping profile, TargetPrint ER, PR and HER2 single gene readout, and the 56-gene TheraPrint Research Gene Panel will be analysed using the whole genome expression array. Patients will receive neo-adjuvant chemotherapy treatment according to protocol. Response will be measured by centrally assessed Residual Cancer Burden (RCB). Objectives are: (1) To determine the predictive power of MammaPrint and BluePrint for sensitivity to neo-adjuvant chemotherapy as measured by pCR. (2) To identify and/or validate predictive gene expression profiles of clinical response or resistance to neo-adjuvant chemotherapy. (3) To compare TargetPrint ER, PR and HER2 with local and centralized IHC and/or CISH/FISH assessment. (4) To identify correlations between TheraPrint and response to neo-adjuvant chemotherapy. (5) To compare BluePrint molecular subtype with IHC-based subtype classification. To achieve a difference of 20% in chemotherapy sensitivity for patients stratified by MammaPrint, a total of 226 samples is needed (significance level 0.05 and power of 0.90). So far 45 patients have been enrolled from multiple institutions. Clinical trial information: NCT01501487.
A pilot study utilizing molecular profiling to find potential targets and select individualized treatments for patients with metastatic breast cancer.
Authors: Jameson Gayle S. , Petricoin Emanuel , Sachdev Jasgit C. , Liotta Lance A. , Loesch David , Anthony Stephen Patrick , Chadha Manpreet , Pierobon Mariaelena , Reeder Alex , Fulk Monica , Vocila Linda , Cantafio Nina , Dunetz Bryant , Robert Nicholas J.
Published: March 2020

TPS11123

Background: The objective of this prospective pilot study was to determine if treatment selected by molecular profiling (MP) of metastatic breast cancer (MBC) tumors at time of disease progression provides greater clinical benefit than empiric treatment selection. Methods: Eligible pts had MBC, ≥ 3 prior lines of therapy, and > 6 wks - < 6 months time to progression on last treatment prior to study entry. Fresh core biopsy samples were taken from the metastatic site for MP. Analysis by IHC, FISH, DNA microarray and reverse phase protein microarray (RPMA) a quantitative protein pathway activation mapping technique using laser capture micro dissected tumor cells was done, with results in ≤14 days reviewed by a Treatment Selection Committee. The primary objective was to compare the progression-free survival (PFS) using a MP selected treatment regimen to the preceding PFS. Clinical benefit for the pt is defined by PFS ratio (PFS on MP selected therapy/PFS on prior therapy) is ≥ 1.3, (JCO,28:4877-83:2010). N= 25 was determined (exact single-stage design for phase II studies, type I error rate of 5% [one-sided], power of 90%). MP selected therapy would warrant further investigation if ≥ 35% of the pts demonstrate a PFS ratio of ≥ 1.3. Results: Three U.S. sites enrolled 25 evaluable pts who were treated based on MP results. Pts were heavily pretreated with 4 -11 prior therapies (median 7.24). Ten pts (40%) met or exceeded the PFS ratio of 1.3. The most common targets used in drug treatment selection were TOPO1, TS, ER/PR, TOP2A, HER2. Sixty percent of pts’ samples had activation of drug targets determined by RPMA in 3 major clusters: pan-HER-AKT; EGFR/SRC/ERK/mTOR; IGFR/RAF/MEK/PLK1. Days on MP selected treatments range from 9 - 816+. 3 pts continue on treatment for 199, 254 and 816 days. Conclusions: This study demonstrates the feasibility of a highly multiplexed genomic and proteomic MP study for treatment selection in a timely fashion. Patient-specific target driven treatment selection based on MP of a metastatic lesion provided clinical benefit for 10 of 25 heavily pretreated MBC pts. Thus, this approach merits further investigation in future studies. Funded by The Side-Out Foundation. Clinical trial information: NCT01074814.
Randomized, open-label, phase II study comparing the efficacy and the safety of cabazitaxel versus weekly paclitaxel given as neoadjuvant treatment in patients with operable triple-negative or luminal b/HER2 normal breast cancer (GENEVIEVE).
Authors: Paepke Stefan , Kümmel Sherko , Blohmer Jens U. , Costa Serban Dan , Eidtmann Holger , Eiermann Wolfgang , Gerber Bernd , Hanusch Claus , Hilfrich Joern , Huober Jens Bodo , Jackisch Christian , Schneeweiss Andreas , Denkert Carsten , Mehta Keyur , Loibl Sibylle , Von Minckwitz Gunter ,   German Breast Group
Published: March 2020

TPS1138

Background: Cabazitaxel is a new taxoid promoting the tubulin assembly in vitro and stabilizing microtubules against cold-induced depolymerization as efficiently as docetaxel. It has shown superior survival against mitoxantrone plus prednisone in docetaxel pre-treated hormone refractory metastatic prostate cancer pts leading to its registration. It showed a favorable toxicity profile with a low rate of alopecia. In the GENEVIEVE study it will be compared to weekly paclitaxel, which is currently most widely used in breast cancer (BC) pts. Methods: This is a prospective multicenter, randomized, open label study investigating efficacy and safety of cabazitaxel. Pts with uni- or bilateral primary BC (stage cT3/T2/T1c and cN+/T1c and pNSLN+), tumor lesion ≥ 2cm (palpation) or ≥ 1cm (sonography) and centrally confirmed TNBC or luminal B/HER2- can be included. Pts will be randomized to four q3w cabazitaxel (25mg/m² i.v.) vs. 12 q1w paclitaxel (80mg/m² i.v.). Randomization will be stratified by nodal status and subtype. Treatment will be given until surgery, disease progression, unacceptable toxicity or withdrawal of consent. The primary objective is pathologic complete response (pCR) (ypT0/is ypN0/+). Secondary objectives are pCR in stratified subgroups and by other definitions, objective response rate, pCR and local recurrence free survival in pts with clinical complete response and neg. core biopsy before surgery, breast conservation rate, toxicity, compliance, survival rates, biomarkers predicting response. Assuming 15% pCR in controls and targeting a smallest clinical improvement of 10% (i.e. pCR = 25% in experimental arm), a total of 326 pts (163/arm) are required for the one-sided proportion comparison test (α=0.1) with 80% power. The trial is registered under NCT01779479. Results: Recruitment is planned for 12 mths in 45 (+10 back-up) sites in Germany. 1st pt in is planned for Feb 2013. Conclusion: GENEVIEVE will rapidly and precisely compare efficacy and tolerability of cabacitaxel vs. paclitaxel to decide if further development in BC is reasonable. Clinical trial information: NCT01779479.
Phase I study of VGX-100, an anti-VEGF-C monoclonal antibody, with or without bevacizumab, in patients (pts) with advanced solid tumors.
Authors: Falchook Gerald Steven , Desai Jayesh , Leitch Ian M. , Goldman Jonathan Wade , Wheler Jennifer J. , Fu Siqing , Kurzrock Razelle , Baldwin Megan , Rosen Lee S. ,   UCLA Hematology-Oncology, Santa Monica, CA; University of Texas MD Anderson Cancer Center, Houston, TX; Royal Melbourne Hospital, Melbourne, Australia; Vegenics, Melbourne, Australia
Published: March 2020

TPS2619

Background: The vascular endothelial growth factor C (VEGFEC) induces angiogenesis via activation of both VEGFRE2 and VEGFRE3, as well as lymphangiogenesis via activation of VEGFRE3. VGX-100 is a novel fully human IgG1λ neutralizing monoclonal antibody directed against human VEGFEC. Synergism between the VEGF-A inhibitor, bevacizumab and VGX-100 has been documented pre-clinically. It is thought that tumoral escape and relapse following VEGF-A inhibition, may in part be due to increased VEGF-C that signals through VEGFR-2 and VEGFR-3. Therefore the premise of this study is that administration of VGX-100 in conjunction with bevacizumab would block the two key ligands for VEGFRE2 along with blocking VEGFR-3-mediated tumor angiogenesis and lymphangiogenesis, and thus this drug combination will be clinically synergistic. This study (NCT01514123) is enrolling at MD Anderson and UCLA. Methods: Objectives:establish the safety and toxicity, MTD, pharmacokinetic and pharmacodynamic / biomarker profiles, as well as preliminary anti-tumor activity in refractory pts. Eligibility: pts with advanced solid tumors, good organ function, ECOG PS 0-1, any number of prior therapies, no CNS or cerebrovascular haemorrhage, no MI or reversible posterior leukoencephalopathy syndrome associated with prior anti-VEGF/anti-VEGFR therapy. Design: This is an open-label phase I dose escalation study with a standard “3+3” design. The study has two arms: safety data from the 28 day DLT period from Arm A (VGXE100 monotherapy at 6 cohort dose levels: 1, 2.5, 5, 10, 20 and 30 mg/kg, QW) will be available prior to starting the equivalent dose level in Arm B (VGXE100 at 5 cohort dose levels: 2.5, 5, 10, 20 and 30 mg/kg QW with bevacizumab at doses of either 5 or 10 mg/kg, Q2W). Accrual has completed in Cohorts A1 to A5 and B1 to B2. Accrual is underway for Cohort A6 and B3. Clinical trial information: NCT01514123.