bluuebunny/crossref_metadata_2025 huggingface dataset:
Association of hypoxia inducible factor-1 alpha and carbonic anhydrase IX overexpression and survival in HER2/neu positive hormone-unresponsive breast cancer patients
Authors: Kaya A. O. , Gunel N. , Benekli M. , Akyurek N. , Buyukberber S. , Tatli H. , Coskun U. , Yildiz R. , Yaman E. , Ozturk B.
Published: August 2020

e22078

Background: Hypoxia is common in solid tumors with overexpression of hypoxia inducible factor-1 alpha (HIF-1 alpha) and carbonic anhydrase IX (CA IX). In our study, we evaluated HIF-1alpha and CA IX expression in Her-2/neu positive hormone- unresponsive breast cancer patients. Methods: One hundred and eleven patients with operable breast cancer were identified who had paraffin-embedded tissue blocks available. Estrogen and progesterone receptors and Her-2/neu expression were evaluated by immunohistochemistry (IHC). FISH was used for IHC 2+ cases. We assigned two groups with 56 patients who have ER and PR receptor positive, Her-2/neu negative (group 1) and with 55 patients who have ER and PR negative, Her-2/neu positive (group 2). HIF 1 alpha and CA IX expressions were investigated and results were compared for both groups. Additionally, we investigated the association between HIF 1 alpha and CA IX expressions with stage, grade, lymph node metastasis, tumor dimension and menopause status. Results: Median age of the patients was 52 years (range, 34–77) in group 1 and 47 years (range, 27–83) in group 2. Nuclear HIF-1alpha expression was detected in 26 (46.4%) and 46 patients (83.6%) in group 1 and 2, respectively (p<0.0001). Membranous CA IX expression was detected in 25 (46.4%) and 37 (67.3%) patients in group 1 and 2, respectively (p<0.013). In group 1, median disease free survival (DFS) has not been reached. In group 2, median DFS was 46 months (95%CI: 42.98–49.02) (p= 0.0308). In group 1, median overall survival (OS) was 108 months (95%CI: 90.65–125.35) whereas in group 2, median OS was 75 months (95 % CI: 64.94- 85.06) (p= 0.0339). Conclusions: HIF-1 alpha and CA IX overexpressions were more frequently observed in ER and PR negative, Her-2/neu positive breast cancer. This was associated with shorter disease-free and overall survival.

No significant financial relationships to disclose.
Profiling of receptor tyrosine kinases (RTK) activation in metastatic breast cancer lesions via proximity mediated microarray immunoassay
Authors: Kim P. , Liu L. , Liu X. , Harvie G. , Zhang L. , Kirkland R. , Magonova K. , Ybarrondo B. , Singh S.
Published: August 2020

e22095

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

[Table: see text]
Treatment related changes of the serum epidermal growth factor receptor in advanced colorectal cancer
Authors: Spindler K. G. , Olsen D. , Brandslund I. , Jakobsen A.
Published: August 2020

e22096

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

No significant financial relationships to disclose.
Expression of CD133, a possible marker for cancer stem cells (CSCs), in small cell lung cancer (SCLC) cell lines and non- small cell lung cancer (NSCLC) cell lines
Authors: Hayashi T. , Tao H. , Jida M. , Kubo T. , Yamamoto H. , Otani H. , Sano Y. , Toyooka S.
Published: August 2020

e22100

Background: Cancer stem cell (CSCs) are believed to play important roles in tumor development, recurrence or metastasis. Identification of CSCs may have a therapeutic significance. CD133 expression has been shown on a minority of various human cancer cells with high capability of self-renewal and proliferation. Therefore, CD133 is thought to be one of possible markers for CSCs. Regarding human lung cancers, the existence, prevalence or roles of CD133 positive cells has not been fully understood. Methods: We examined CD133 mRNA by quantitative real-time PCR and sorted CD133-positive cells by fluorescence-activated cell sorting (FACS) using human small cell lung cancer(SCLC) and non-small cell lung cancer (NSCLC) cell lines. We evaluated differences of cell proliferation between CD133-positive and -negative cells by MTS assay in vitro and by subcutaneous injection for non- obese diabetic/severe combined immunodeficiency (NOD/SCID) mice in vivo. Results: CD133 expression was almost restricted in SCLC cell lines. CD133 mRNA expression or CD133-positive cell population was scarcely observed in NSCLC cell lines. In two SCLC cell lines examined (NCI-H82 and NCI-H69), CD133 positive cells had higher tumorgenicity both in vivo and in vitro than NSCLC cell lines. Conclusions: The expression status of CD133 is totally different between NSCLCs and SCLCs, probably reflecting the difference of these progenitor cells. Our results indicate that CD133-positive cells in SCLC cell are responsible for tumor growth. However, in view of their wide prevalence, CD133-positive cells do not seem to be a candidate for CSCs, at least in cell lines. To investigate the molecular and functional characteristics of CD133-positive cells may lead to a new therapeutic strategy for human lung cancers, especially for SCLCs.

No significant financial relationships to disclose.
Soluble vascular endothelial growth factor receptor 2 (VEGFR2): New biomarker in advanced non-small cell lung cancer (NSCLC)?
Authors: Sanmartin E. , Jantus Lewintre E. , Sirera R. , Miñana M. , Navarro A. , Cabrera A. , Blasco A. , Pla A. , Rosell R. , Camps C.
Published: August 2020

e22108

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

No significant financial relationships to disclose.
Integrating microRNA and mRNA expression profiling using a novel algorithm identified a small set of unique genes upregulated in malignant pleural mesothelioma (MPM)
Authors: Suraokar M. , Corvalan A. , Chow C. , Gazdar A. , Moran C. , Raso G. , Mehran R. , Tsao A. , Wistuba I.
Published: August 2020

e22111

Background: We employed a global profiling strategy using miRNA microarrays in MPM cell lines and archival tumor tissue. Methods: We isolated total RNA from 4 MPM cell lines, 2 control cell lines, and 16 tissue specimens from patients with resected MPM (n=8) and normal counterpart (n=8) patients as controls. Total RNA was labeled with Cyanine 3, then hybridized with Agilent human miRNA microarray v1 slides. Results: Preliminary miRNA profiles show up-regulation of 44 versus down-regulation of 29 miRNA's in MSTO-211H cancer cells compared to HCT-4012 (pleural telomerase-transformed control). Profiling of 16 tissue specimens (8 normal vs 8 MPM) revealed down-regulation of 11 miRNA's in MPM tumor tissue. To focus on relevant miRNA that regulate genes involved in carcinogenesis and progression, we identified > 1000 unique genes using the online targetscan 4.2 program ( http://www.targetscan.org ), which predicts biological targets of miRNAs by identifying the presence of conserved 8-mer and 7-mer sites that match the seed region of each miRNA. We then explored a novel screening strategy, which combines mRNA expression dataset with the miRNA dataset, to narrow the list of relevant miRNA's. We conducted gene expression profiling on the cell lines and MPM tissue samples with Affymetrix U133 plus 2.0 chips. Bioinformatic analysis was conducted with MeV: MultiExperiment Viewer software, data reduction techniques (Correspondance Analysis), hierarchical clustering methods, and Serial Analysis for Microarray (SAM), and showed up-regulation of ∼300 genes in MPM compared to normal tissues. We then computed of the ∼300 mRNA's up-regulated in MPM only 32 are recognized by the 11 down-regulated miRNA's using the targetscan 4.2 algorithm. Most of the miRNA's regulate single messages while ∼20 % of the messages are regulated by more than 1 miRNA's. Some of these targets include Ets variant 1 and Protein kinase C - epsilon. Conclusions: This innovative approach of selecting highly relevant miRNA is feasible and enables discovery of novel genes based on their ability to be bound by single or multiple miRNA's. Validation of our profiling studies using real-time PCR and protein analysis methods will be presented.

No significant financial relationships to disclose.
Cancer incidence in HIV-positive patients in Germany: A nation-wide survey from 2000 to 2007
Authors: Hensel M. , Goetzenich A. , Hanhoff N. , Wolf E. , Knechten H. , Mosthaf F.
Published: August 2020

e22115

Background: Malignancies are an essential feature of acquired immunodeficiency syndrome and human immunodeficiency virus (HIV) infection. The purpose of this study was to gather data on the epidemiology of AIDS-defining (AD) and non-AIDS-defining (NAD) malignancies in HIV-positive patients (pts) in Germany in the past decade. Methods: Study centers (all HIV-specialty clinics and ambulatory care centers in Germany, all members of the German association of medical oncologists in private practice) were contacted annually between 2000 and 2007 and asked to respond to a structured questionnaire. The questionnaire requested information on all malignancies in HIV-positive pts, tumor stage, CDC (Center for Disease Control)-stage of the HIV infection, sex, treatment and clinical course. Results: 111 centers participated in the evaluation and provided 552 evaluable data sets from 542 pts. 89% of cases were male. The majority of pts had advanced HIV-disease (CDC stage C3), but the proportion of pts with stage C3 decreased from 58% in 2000 to 36.8% in 2007. 253 (45.8%) were AD as follows: 132 Kaposi Sarcomas, 109 aggressive B-cell lymphomas, 12 invasive cervix carcinomas. The B-cell lymphomas further included 28 Burkitt's lymphomas, 30 DLBCL, 9 Castleman diseases, 8 primary cerebral lymphomas. Among the 299 cases (54.2%) of NAD malignomas were 213 solid tumors including 71 anal carcinomas (= 33.5% of all NAD malignancies) and 85 hemoblastoses including 29 Hodgkin lymphomas (= 9.6% of all NAD malignancies). The high proportion of NAD malignancies has remained constant over all observation periods, as well as the relative incidence of most of the different subentities. Interestingly, only 1 of 8 primary cerebral lymphomas has been reported after 2001. The number of pts with Hodgkin's lymphoma has increased constantly from 2000 to 2007. Conclusions: Our observations show a high incidence of NAD malignomas over the past 8 years in Germany. Anal carcinomas and Hodgkin's lymphomas in particular were markedly more prevalent in our HIV-positive cohort compared to published reports of the general population. The incidence of primary cerebral lymphomas seems to decrease, whereas the incidence of Hodgkin's lymphoma is increasing.

No significant financial relationships to disclose.
Circulating tumor cell detection: A direct comparison between the CellSearch System, the AdnaTest, and CK-19/mammaglobin RT-PCR in patients with metastatic breast cancer
Authors: Dirix L. Y. , Elst H. , Benoy I. , Van der Auwera I. , Prové A. , Wuyts H. , Maes H. , van Dam P. , Huget P. , Vermeulen P.
Published: August 2020

e22117

Background: The detection, enumeration and isolation of circulating tumor cells (CTC) has considerable potential to influence the clinical management of patients with breast cancer. There is however a substantial variability in the rates of positive samples using existing detection techniques. Methods: This study was designed to compare three techniques for detecting CTC in blood of 80 patients with metastatic breast cancer (MBC) and 20 healthy controls: the CellSearch System, which is an automated, standardized and regulatory-approved system for the immunocytochemical detection and quantification of CTC in blood; ii) the AdnaTest Breast Cancer Select/Detect, which involves the detection of tumor-associated transcripts by RT-PCR after an immunomagnetically enrichment of tumor cells; iii) an in-house developed multimarker qRT-PCR assay, which involves the quantification of tumor-associated transcripts (CK-19 and MAM) by qRT- PCR. Results: As a result, 23% of patients with MBC were positive by the CellSearch System (≥5 CTC), 22% by the AdnaTest (>0.30 ng/μl for any of the amplicons), 31% by qRT-PCR for CK-19 and 49% by qRT-PCR for MAM. Samples were more likely to be positive by qRT-PCR for at least one mRNA marker than by the CellSearch System (P<0.001) or the AdnaTest (P <0.001). The concordance between samples analyzed by the CellSearch System and the AdnaTest was substantial (κ = 0.667, P <0.001). Agreement between both detection techniques was observed in 88% of blood samples. When the CellSearch System was compared with the qRT-PCR assays for CK-19 and MAM, we observed agreement percentages of 78% and 58%, respectively (κ = 0.462, P <0.001 and κ = 0.159, P = 0.09). Agreement between the AdnaTest and the qRT-PCR assays for CK-19 and MAM was observed in 78% and 53% of blood samples, respectively (κ = 0.443, P <0.001 and κ = 0.05, P = 0.607). Conclusions: We observed a substantial variation in the detection rates of CTC in blood from MBC patients using three different techniques. A higher rate of positive samples was observed using a combined qRT-PCR approach for CK-19 and MAM, which suggests that this is currently the most sensitive technique for detecting CTC.

No significant financial relationships to disclose.
Neuroendocrine tumors in children and young adults: Incidence, survival, and prevalence in the United States
Authors: O'Dorisio M. S. , Navalkele P. , O'Dorisio T. M. , Lynch C. F.
Published: August 2020

e22120

Background: Neuroendocrine tumors (NET) arising from the diffuse endocrine system are thought to be quite rare in children and young adults. However, a surprising number of young people have been referred to our neuroendocrine tumor clinic and the NCI has targeted NET as a high priority for development of new diagnostic and therapeutic options. This analysis of the SEER database was undertaken to determine the incidence, prevalence, and survival of NET in young people. Their incidence, prevalence, and survival were compared with neuroblastoma, a related pediatric malignancy arising in the neural crest. Methods: The SEER data were obtained from 9 standard SEER registries for the diagnosis years of 1975 to 2004 using SEER*Stat version 6.4.4. ICD-9 codes related to neuroendocrine tumors and to neuroblastoma were characterized as to patient age, gender, racial and ethnic background, stage, grade, histology, incidence, survival, and prevalence. Results: Neuroendocrine tumors occur more often in females among children and young adults with the most common sites being bronchial, ovarian, and breast. The overall incidence of neuroendocrine tumors was lower than for neuroblastoma in the age range 0–30 years. However, the 30 year limited prevalence of neuroendocrine tumors in the 9 SEER registries was 698 compared to 881 for neuroblastoma. This extrapolated to over 7000 children and young adults with neuroendocrine tumors across the United States. Survival rate of young people with neuroendocrine tumors declined from 84% in 1975–1986 to 80% in the 1987–2004 era. Conclusions: These results indicate that neuroendocrine tumors constitute an unrecognized cancer threat to children and young adults. Survival of children and young adults with neuroendocrine tumors has decreased over the past 30 years in the United States. We recommend the establishment of centers of care for children and young adults diagnosed with neuroendocrine tumors with the expectation that earlier diagnosis coupled with targeted therapies will decrease the incidence of metastatic disease and improve survival.

No significant financial relationships to disclose.
Relationship of systemic immune dysregulation in advanced tumors to vascular growth factor overproduction
Authors: Block M. S. , Markovic S. N.
Published: August 2020

e22124

Background: Therapeutic anti-cancer vaccine strategies have been effective in a broad range of preclinical tumor models, but clinical trials of vaccines in metastatic cancer patients have failed to show clinical benefit. We have previously shown that patients with stage IV melanoma have systemic immunosuppression including elevated plasma levels of immunosuppressive cytokines and impaired DTH responses to recall antigens. Patients with stage IV melanoma also have elevated circulating levels of the vascular growth factors vascular endothelial growth factor A (VEGF A) and placental growth factor (PlGF). Based on these observations, we have hypothesized that vascular growth factors such as VEGF and PlGF mediate systemic immunosuppression. Methods: FVB mice with established ANV6 transplantable breast tumors were bled, and plasma samples were assayed for VEGF, PlGF, and interleukin (IL)-10. Splenocytes taken from tumor-bearing mice were stimulated via the T cell receptor (TCR), and interferon gamma (IFN) production was measured. Splenocytes from normal mice were stimulated in the presence or absence of VEGF; IFN production was measured. Results: Compared with age- and sex-matched controls, tumor-bearing mice had elevated plasma levels of VEGF, PlGF, and the immunosuppressive cytokine IL-10 (see table ). Furthermore, splenocytes from tumor-bearing mice produced less IFN upon TCR stimulation than did splenocytes from control mice. Finally, normal mouse splenocytes produced less IFN when stimulated in the presence of VEGF than in its absence (70.2 ± 37.2 vs. 260.0 ± 130.9 pg/ml). Conclusions: We have shown that mice with advanced tumors exhibit both elevated levels of vascular growth factors and systemic immune inhibition. We have also shown that VEGF leads to impairment of immune function in normal splenocytes. Taken together, these results suggest that excess vascular growth factors may mediate immunosuppression in animals with advanced tumors, which may in turn impair anti-tumor vaccine efficacy.

[Table: see text]

No significant financial relationships to disclose.
Are more genes better in clinical-genomic studies? A mathematical model to define identification power for clinically relevant genes
Authors: Crager M. R. , Shak S.
Published: August 2020

e22136

Background: Modern molecular technologies that drive personalized medicine can generate expression data for thousands of candidate genes, or indeed, data for the “whole genome”. Clinical-genomic studies aim to identify genes that are truly associated with clinical outcome. We investigated the impact of large numbers of genes with little or no association with clinical outcome on the statistical power of studies to identify individual genes with strong association. Methods: We adapted Efron's (Ann Stat 2007) empirical Bayes approach to develop a method to calculate the identification power for individual genes in analyses that control the false discovery rate (FDR), the expected proportion of false positives. The identification power is the probability that a gene with a given true magnitude of association with clinical outcome will be identified when we control the FDR at a specified level. The identification power also depends on the proportion of genes studied that are not associated with outcome and the distribution of the degree of association among genes that are. Results: The identification power for clinically relevant genes decreases dramatically as the proportion of genes having no association with clinical outcome increases. For example, in a scenario in which 100 genes have some association with clinical outcome [median hazard ratio (HR) 1.125], increasing the number of genes having no association from 400 to 4000 decreases the identification power an individual gene having strong association (HR 1.42) from 80% to 36%. Similarly, when the number of genes having no association is 400 and the median HR among the 100 genes that have an association is decreased from 1.125 to 1.06, identification power for a gene with an association HR of 1.42 decreases from 80% to 62%. Conclusions: Identification power can be used to optimize strategies for gene finding and enable personalized medicine. Although technology allows the assay of ever-increasing numbers of genes, inclusion in a single study of many genes unrelated to clinical outcome is detrimental to the identification power for clinically relevant genes. Even with increased power, replication of results in independent studies will continue to be critically important.

[Table: see text]
Selection of lung cancer-associated scFv antibodies from cultured cells
Authors: Massion P. P. , Pedchenko T. V. , Parekh D. V. , Mernaugh R.
Published: August 2020

e22137

Background: Lung cancer is the most common cause of cancer-related deaths in the world. There is a critical need for new strategies of early lung cancer detection. The identification of tumor-associated antigens and corresponding antibodies is one approach to discovery of diagnostic biomarkers. We used a large phage-displayed recombinant antibody library and normal human lung epithelial and non-small cell lung cancer cell lines to select for and identify recombinant antibodies specific for proteins expressed, or over-expressed, in lung cancer. Methods: The antibody library was used to select for recombinant scFv antibodies reactive with proteins present, or aberrantly expressed, in non-small cell lung cancer cell lines (A549, H549, H157, H23) in comparison to normal lung cell lines (BEAS-2B, 16-HBE, KT). Soluble scFv antibodies were obtained through 2 rounds of phage antibody cross-absorption (on normal cell lines) and selection (on non-small lung cancer cell lines). Soluble scFv were assayed by a high-throughput fluorometric microvolume assay technology (FMAT) against normal and cancer lung cell line proteins. ScFv antibodies selected by FMAT were evaluated further with Western blot-based assays. Results: More than 100 scFv antibodies identified by FMAT bound preferentially to proteins in lung cancer. Of these, 46 scFv were assayed by a high throughput Western slot blot immunoassay against pooled normal lung and lung cancer cell lysates. Eight scFv were assayed in Western blot against individual lung normal and non-small lung cancer cell line lysates. Four of these demonstrated differential binding to normal and cancer cell lysates. Conclusions: In summary, we were able to detect cancer-associated antigens in lung cancer cell lines using a phage display antibody library. In combination with high-throughput fluorescent and Western blot assays, four unique scFv antibodies were selected that differentially bound to normal and lung cancer cell lysates. These scFv will be tested as candidate biomarkers of lung cancer in independent tissue and serum samples from patient with and without lung cancer to determine utility for use in lung cancer diagnosis.

No significant financial relationships to disclose.
The oncologist as genetic consultant: Two-year results in a large community-based practice
Authors: Geier L. J. , Sheehan M. , Elia M. , Ethirajan S. , Kent E.
Published: August 2020

e22140

Background: Oncologists are in a unique position to provide effective genetic cancer risk assessment (GCRA). However, in most communities they defer this to genetic counselors (GC). At KC Cancer Center, a 35-MD community oncology group, the difficulty obtaining timely and effective GCRA became a quality of care issue. Methods: In December 2006 a new program was launched with 4 designated oncologists providing GCRA at 4 separate cancer centers. Each oncologist was responsible for gaining expertise in the common hereditary syndromes and appropriate interpretation of gene testing results prior to starting GCRA. Two-year results were analyzed. For those pts found to have deleterious mutations, charts were reviewed to confirm the gene affected, and the length of time between cancer dx and GCRA. Pts more than one year out from dx were designated as cancer survivors. Results: After 2 years, 490 pts had gone through gene testing after GCRA, and 85 were found to carry deleterious mutations. These data were compared to the previous three years when pts had been referred to GC ( Table ). Both the number of pts tested and the number with mutations in yrs 2007–8 were approximately 400% of the numbers for yrs 2005–06. Of the 85 pts with mutations, 51 (60%) were cancer survivors. Their median time from diagnosis was 5.5 yrs (range 1–25). 75/85 (88%) mutations were in a BRCA gene. Conclusions: This oncologist-centered model proved to be very effective in identifying mutation carriers, particularly among cancer survivors in whom the hereditary syndrome had been previously overlooked. Acceptance of this approach by pts, physicians, and payers has been extremely high. This model should be considered by oncology practices wanting to add GCRA to their service lines.

[Table: see text]

[Table: see text]
Novel de novo BRCA1 mutation in a woman with early onset breast cancer
Authors: Tang E. , Kwong A. , Wong C. , Law F. , Wong C. , Ng E. , Ma E. , Ford J. M.
Published: August 2020

e22143

Background: Germline mutations in BRCA1/2 account for a significant portion of hereditary breast/ovarian cancer. Mutation carriers usually have a family history of breast/ovarian cancer or early onset disease. Rarely, germline mutations are found only in the probands but not in any family members. Such de novo mutations have been reported in diseases such as hemophilia A, thalassaemia and familial adenomatous polyposis. De novo mutations in the BRCA1 or BRCA2 genes are rare and the few reported have been in BRCA2. Here, we describe de novo as well as novel mutation of the BRCA1 gene in a breast cancer patient. Methods: Blood DNA samples from a 30 year old Chinese woman with breast cancer and no family history of cancer was tested for a BRCA1/2 mutation by full gene sequencing and Multiple Ligation-dependent Probe Amplification (MLPA). Family members were analyzed for the same mutation. Paternity was determined by a set of highly polymorphic short tandem repeat (STR) markers. Results: Full gene sequencing found no deleterious mutation. MLPA revealed a large deletion of exons 1 to 12 of BRCA1 in the proband. MLPA performed on 5 family members: proband's mother and father (who were 1st degree relative- cousins), stepmother (mother's biological sister), 2 sisters (1, same parents; 1, same father and stepmother) found no similar deletion. By using a set of highly polymorphic STR markers, the proband's father and mother were confirmed to be her biological parents. Conclusions: We report a novel de novo BRCA1 deletion mutation encompassing exons 1 - 12 in a Chinese breast cancer patient of early onset with no family history. Identification of this large deletion confirms the importance of pursuing rearrangement testing if full gene sequencing fails to detect a point mutation or short insertion deletion. The mutation found in this study is de novo. This may simply be a random mutation event which occurred in the parents' germ cells during their lifetime which passed onto one of their offspring or maybe a result of gene inversion or splicing deficiency. The relations of such mutations with consanguineous marriage cannot be ruled out. Mutation screening is important in early onset breast cancer patients even if there is no family history.

No significant financial relationships to disclose.
Survival analysis in breast cancer's women in Santa Catarina, south of Brazil
Authors: Schneider I. J. , Orsi E. D. , Furtado C. M. , Vianna A. , Silva T. S.
Published: August 2020

e22149

Background: Overall survival after breast cancer diagnosis is influenced by the characteristics study's group, the study's place and biological factors. The prognostic factors are directly related to the survival of women with breast cancer. The studies indicate that the incidence and survival of breast cancer are influenced by socioeconomic factors and demographics. Methods: Define overall survival in five-years and the associated factors with the prognosis of breast cancer's women in Santa Catarina, Brazil. This is a historic cohort study, with data from diagnoses from January 2000 to December 2002 from Registro Hospitalar de Câncer of the Centro de Pesquisas Oncológicas, SC and the Hospital de Caridade, both in south Brazil, and complemented with data from the Mortality Information System until December 2007. The time of survival was calculated as the range of the date of diagnosis (biopsy or surgery) until the date of death or end of monitoring (5 years). The survival curves were estimated by the Kaplan-Meier method, using the log-rank test for comparison of the curves. The effect of co-variable model was estimated by the semi- parametric risk proportionate, Model Cox, with the Schoenfelder analysis of residuals to assess whether the co-variables of the final model respect the principle of proportionality over time. The analysis was made using R 2.5.0. Results: 1002 women were included, and 262 deaths occurred, this 235 by breast cancer. The overall survival after 5 years was 76,2% (73,6–78,9%). Variables were associated with survival by the Kaplan-Meier curves: age, race/color, education, clinical stage, treatments received. This includes these variables in the model of Cox, remaining in the final model, as independent factors: age until 30 years (HR: 3,09) illiterate (HR 3,70), staging III (HR 5,27) and staging IV (HR 14,07).The Cox model variables respect the principle of proportionality over time (p=0,25). Conclusions: The overall survival in breast cancer, proved to be better than in other Brazilian states. The instruction presents itself as a strong influence on survival, where illiterate women have worse prognosis. The diagnosis in advanced stages is one of the most important factors for worse survival, and modifiable through campaigns of early detection and mammography screening.

No significant financial relationships to disclose.
Effect of panitumumab on autophagy in colon cancer
Authors: Kalofonos H. P. , Antonacopoulou A. , Matsouka P. , Giannopoulou E.
Published: August 2020

e22151

Background: Panitumumab, a human monoclonal antibody raised against EGFR, has been approved by the Food and Drug Administration (FDA) and the European Medicines Agency (EMEA) for the treatment of patients with EGFR-expressing mCRC and wild type kras. The ratio of reduced/oxidised form of glutathione (GSH/GSSG) is an indicator of the redox status in cells. The aim of the current study was to investigate the effect of panitumumab on the redox status of colon cancer cell lines Caco-2, DLD-1 and HT-29 regarding proliferation, apoptosis, necrosis, cell cycle arrest and autophagy. Methods: Cell proliferation was measured by MTT assay. Apoptosis and necrosis were detected by annexin v/propidium iodide assay. Cell cycle arrest was estimated by propidium iodide assay. Autophagy was detected by immunobloting and GSH levels were measured by spectrophotometrical analysis. kras mutations were detected by sequencing analysis. Results: Caco-2, DLD-1 and HT-29 cell lines differ in the expression levels of EGFR and HER-2. Kras mutation analysis in previous studies and in the current study showed that DLD-1 cells express mutated kras while Caco-2 and HT-29 cells express wild type of kras. Panitumumab decreased proliferation only in DLD-1 cells 48 h after its application besides the mutated kras. However, panitumumab did not affect DLD-1 cell apoptosis, necrosis or cell cycle progression 24 and 48 h after cells treatment. Interestingly, panitumumab increased protein levels of beclin 1, an indicator of autophagy, 24 h after its addition in cells. Moreover, an increase in GSH levels was noted 48 h after cells treatment with panitumumab. Conclusions: This is the first study to show that panitumumab, an EGFR inhibitor, affects colon cancer cell proliferation independently of kras mutations and EGFR protein levels through the induction of autophagy. The inhibition in cell proliferation was followed by an increase in GSH levels reflecting an imbalance on the redox status of cells.

No significant financial relationships to disclose.
N-cadherin, E-cadherin, ERCC1, and c-kit expression in small cell lung cancer (SCLC) and potential for new therapeutic targets
Authors: Batus M. , Myint R. , Coon J. , Basu S. , Kaiser K. , Fidler M. , Bonomi P.
Published: August 2020

e22157

Background: Minimal advances have been made in the treatment of SCLC. Molecular markers may allow us to better stratify patients (pts) for new treatment options and drug combinations. The objective of our study was to determine the frequency and potential prognostic significance of N-cadherin (N-cad), E-cadherin (E-cad), ERCC1, and c-kit (CD117) expression in SCLC. Methods: Tissue from 132 pts with SCLC was retrospectively stained for N-cad, E-cad, ERCC1, and c-kit. Frequency of expression (% of tumor cells staining positive) was measured on a scale of 0–4 (freq 0=no expression (<1%), freq 1=1–10%, freq 2=11–35%, freq 3=36–70%, freq 4=71–100%). Charts were reviewed for stage, performance status, date of diagnosis/death, survival, and treatment (type, dates, response). The frequency of molecular markers was correlated with clinical data and overall survival. Results: Age range 42 to 97 years, 65 male:67 female, and 64 had limited and 68 had extensive stage. Of the 132 pts, 75% had tumors that expressed (frequency ≥ 1) N-cad, 58% E-cad, 70% ERCC1, and 55% c-kit. Comparing tumor marker expression with survival using either the Log-Rank Test or the Wilcoxon Test, there was no significant association for N-cad, E-cad, or ERCC1. However, tumors that expressed c-kit with frequency ≥ 3 had a trend toward superior survival compared with frequency < 3. Median survival for c-kit frequency ≥ 3 was 496 days compared to 312 days for frequency < 3 (p = 0.09, Log-Rank Test). Conclusions: In our retrospective study of 132 SCLC pts, we found that all 4 markers were expressed in greater than 50% of specimens, and that higher c-kit expression was associated with marginally significant increase in overall survival. Though previous experience with imatinib alone or with chemotherapy showed limited clinical activity in unselected SCLC pts, given preclinical synergy with cisplatin, it seems reasonable to consider combination therapy with cisplatin/etoposide and imatinib in pts selected for high c-kit expression.

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No significant financial relationships to disclose.
Evaluation of estrogen receptor expression as a prognostic factor for very young women (below 35 years) with breast cancer in Taiwan
Authors: Lin C. , Huang C. , Lu Y. , Laiu J. , Huang S. , Kuo W. , Kuo W. , Chang K. , Cheng A.
Published: August 2020

e22163

Background: In the past two decades, the incidence of very young (<35 years) breast cancer in Taiwanese has been rapidly increasing to quite close to that of Caucasian Americans. However, the clinicopathological features of this group of patients appear to be different from their Western counterpart. Here, we sought to determine the prognostic value of ER expression for this newly emerging population in Taiwan. Methods: We retrospectively collected information on all consecutive very young patients (age <35 years) with stage I-III breast cancer treated at National Taiwan University Hospital between 1997 and 2005. ER expression was evaluated by immunohistochemistry on formalin-fixed paraffin-embedded tissue sections. Tumor with more than 10% positive nuclei was considered ER positive. Results: A total of 178 patients with a median age of 32 (range: 21–35) years were included. The median follow-up time was 62.5 (95% confidence interval, 42.0–64.2) months. Univariate analysis showed that ER expression was significantly associated with favorable OS (HR=0.48, P=0.03), but not with DFS (hazard ratio [HR]=0.79, P=0.42). Multivariate analysis showed that stage and ER were independent prognostic factors. Conclusions: In contrast to the findings in Caucasians, ER expression was an independent prognostic factor for favorable overall survival in Taiwanese patients with very young breast cancer.

No significant financial relationships to disclose.
Breast cancer ErbB2 status in Asian women: A review of local literature
Authors: Tan Y. , Han S. , Lu Y. , Yip C. H. , Sunpaweravong P. , Jeong J. , Caguioa P. B. , Aggarwal S. , Yeoh E. M. , Moon H.
Published: August 2020

e22164

Background: Breast cancer incidence is increasing throughout Asia. However, the characteristics of Asian breast cancer are not always well understood due to the limited availability of local data and their inadequate inclusion in global literature databases. Effective breast cancer treatment in this region requires a better understanding of Asian breast cancer biology, including ErbB2 status. Methods: We performed a literature search in seven Asian countries on breast cancer studies in which tumour ErbB2 overexpression was assessed. The keywords erbB2 OR HER2 OR ErbB-2 OR HER-2 AND breast cancer AND (country) were used to search PubMed, international and local conference abstracts and local-language journals from the year 2000 onwards. Where available, we selected up to five representative studies from each country on the basis of population size, multi-institution patient populations, institution reputation and journal impact factor. We excluded studies containing only mRNA, serum, or cell line data or those conducted on biased populations or in western countries on Asian populations. Results: ErbB2 data from 26 Asian breast cancer studies are summarized in the table . The mean or median age of the study populations ranged from 46–56 years, with one exception. In most studies that evaluated tumour ErbB2 and hormone status, ErbB2 over-expression correlated negatively with ER positivity. Conclusions: The larger studies in particular confirm that the proportion of ErbB2-positive breast cancer in Asia is generally similar to the 20–30% reported for western women. Definitions of ErbB2 positivity using IHC vary between institutions and FISH is not routinely performed in several Asian countries. This may contribute to the fact that reliable, representative data on breast tumour ErbB2 status is scarce in Malaysia, the Philippines, India and Thailand. The increased availability of accurate ErbB2 testing and data would aid improved treatment of ErbB2-positive breast cancer in these countries.

[Table: see text]

[Table: see text]
COX-2 expression and clinical outcome in early-stage breast cancer patients treated with adjuvant chemotherapy: A prospective study
Authors: Kargi A. , Ozdogan M. , Bozcuk H. , Pestereli E. , Artac M. , Coskun H. S. , Mutlu H. , Uysal M. , Karaveli S. , Savas B.
Published: August 2020

e22165

Background: To evaluate the association of cox-2 expression with the outcome after adjuvant chemotherapy in patients with early breast cancer. Methods: This was planned as a prospective study recruiting consecutive patients receiving adjuvant anthracycline based chemotherapy and with available tissue blocks permitting all immunohistochemical analyses. Cox-2 expression, in addition to other classical biological factors, was evaluated with immunohistochemistry. Disease and patient related, and biological predictors of both overall survival (OAS) and relapse free survival (RFS) were analyzed by Cox regression analysis. Median and mean survival times were calculated according to the Kaplan Meier method. Results: A total of 88 patients were recruited over a period of 24 months. Median age was 45 (29 to 70), and 60% of subjects were premenapausal. Median tumour diameter and number of axillary lymph nodes involved were 2 cm (1 to 6 cm), and 2 (0 to 15), respectively. Median follow up is 74.2 months. Univariate analysis revealed menopausal status and estrogen receptor expression as predictors of OAS, and menopausal status as the correlate of RFS. Multivariate analysis confirmed the independent predictive value of both menopausal status and estrogen receptor expression for OAS (P=0.009, HR=4.18, and P=0.014, HR=0.20, respectively). No multivariate analysis could be performed for RFS. Cox-2 expression was not associated with OAS or RFS (P=0.208, HR=1.92, and P=0.132, HR=1.89, respectively). Interestingly, Cox-2 expression was correlated with Estrogen receptor (ER) and Progesteron receptor (PR) expression (P=0.006, R=-0.303, and P=0.004, R=-0.312, respectively). Conclusions: Cox-2 expression fails to predict clinical outcome of early breast cancer patients treated with adjuvant chemotherapy. However, Cox-2 expression seems to negatively correlate with ER and PR expression. It should be tested in this patient population whether Cox-2 may play a part in hormonal resistance.

No significant financial relationships to disclose.