bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of expression of TGF-β1 receptor (ALK-1) in normal and tumor tissues by tissue microarrays
Authors: Erbersdobler A. , Simon R. , Hellwinkel O. J. , Bokemeyer C. , Sauter G. , Hu-Lowe D. , Levin W. , Gallo-Stampino C. , Fiedler W. 
Published: August 2020

e22044 Background: TGF-β1 is an important angiogenic factor involved in different aspects of angiogenesis and vessel maintenance. TGF-β1 receptors consist of class I, class II and accessory receptors. Activin-like kinase I (ALK-1) is a class I TGF- β1 receptor which is almost exclusively expressed on endothelial cells. Hereditary hemorrhagic telangiectasia (HHT) is caused by mutations in the ALK-1 gene. Knock-out mice for ALK-1 die during gestation due to vascular malformation. ALK-1 may thus represent an attractive target for anti-angiogenic therapy. A therapeutic monoclonal antibody (PF-03446962) against ALK-1 has been developed which is currently in phase I testing. Here we describe target validation in normal and tumor tissue using tissue micro-arrays (TMAs). Methods: Normal tissue arrays comprised of 609 individual histological samples representing 76 human tissues. Multi-tumor arrays consisted of 3923 individual tumor samples from 87 tumor types. Consecutive sections of tissue arrays were stained for ALK-1 and CD31. CD31 immunohistology (IHC) was performed to ensure presence of vessels within tissue samples. ALK-1 staining was graded 0=absent, 1=weak, 2= moderate and 3=strong by an experienced pathologist blinded to tissue type. Results: ALK-1 staining of vessels in normal tissues was generally weak (mostly grade 1 and rarely grade 2) and was detectable in lymphatic tissues including tonsil, lymph nodes, thymus and spleen, lung, the entire GI tract including parotid, submandibular and sublingual glands as well as pancreas. ALK-1 positive vessels were also found within the female genital tract including placenta, uterus and ovary. In human tumor vessels ALK-1 expression showed high variability between tumor types. The highest ALK-1 expression rate was found in lung cancer (NSCLC 49%, SCLC 83%), neuroendocrine pancreas tumor (71%), colon cancer (50%), chondrosarcoma 50%, angiosarcoma 40% and NHL (44%). Conclusions: TMAs are an excellent tool to verify target expression in normal and tumor tissues. Results may be used to predict side effects of targeted therapy and to direct phase II testing of innovative agents in patients likely to respond. [Table: see text]
Treatment of VHL patients with sunitinib: Clinical observations and translational studies
Authors: Matin S. F. , McCutcheon I. E. , Gombos D. S. , Waguespack S. G. , Wen S. , Smith L. A. , Zhang Y. , Davis D. W. , Fuller G. , Jonasch E. 
Published: August 2020

e22047 Background: Von Hippel Lindau (VHL) disease induces vascular lesions in multiple organs. Surgical intervention is the treatment of choice. Effective, safe systemic therapy will greatly improve the quality of life of these individuals. Methods: Patients with genetically confirmed VHL were enrolled on an IRB approved trial. Eligibility criteria included retinal angiomas, hemangioblastomas (HBs) measuring at least 5mm, renal cell carcinoma (RCC) 1 to 3 cm and pancreatic neuroendocrine tumors (NETs) 1 to 3 cm. Patients received sunitinib 50mg/day for 28 days/14 days off for four cycles. Scans were performed at baseline, and after cycles two and four. A separate set of 20 formalin-fixed paraffin embedded HBs and 20 RCCs were used for laser scanning cytometry (LSC) analysis of total and phospho vascular endothelial growth factor receptor (pVEGFR) and phospho platelet derived growth factor receptor (pPDGFR) levels in tumor endothelium. Results: Twelve patients were evaluable for response and toxicity. Eight had RCCs, nine had CNS lesions, and three had pancreatic NETs. Tumor size reduction was seen in 16/19 evaluable RCC lesions (95%CI 0.60, 0.97), 3/5 NETs (95%CI 0.15, 0.95) and 6/19 HBs (95%CI 0.13, 0.57). Two patients came off study for treatment related side effects, one patient progressed on study, and two patients did not complete the scheduled course of therapy. LSC analysis demonstrated significantly lower phospho VEGFR levels in HBs when compared with RCC. The mean (SD) of pVEGFR levels in log-transformation were 11.27 (0.49) and 11.75 (0.37) for HBs and RCC respectively (p = 0.003). The mean (SD) of pVEGFR to VEGFR ratio was .21 (0.12) versus 0.37 (0.43), for HBs and RCC respsectively (p = 0.043). Conclusions: Sunitinib treatment of patients with VHL resulted in consistent decrease of RCC and NET tumor size. The discrepant response to sunitinib in RCC and HBs may be explained by differential dependence on VEGFR activation in tumor endothelium. [Table: see text]
Expression of stem cell markers in circulating melanoma cells
Authors: Fusi A. , Busse A. , Ochsenreither S. , Rietz A. , Keilholz U. 
Published: August 2020

e22056 Background: Within circulating tumor cells there may be a subset of cell with stem cell (tumor initiating) characteristics able to develop distant metastasis. Several markers including nestin and CD133 have been found to be possible candidates to identify such a kind of subpopulation in other experimental models. We evaluated the presence of melanoma cells bearing stem cell phenotype in the bloodstream of patients with cutaneous or uveal melanoma after depletion of the leukocytes fraction. Methods: Between 50 and 100 ml of peripheral blood were collected from 12 melanoma patients with various tumor burden as well as three healthy volunteers. Blood samples were enriched for tumour cells by CD45 depletion of the leukocyte fraction using magnetic beads separation (EasySep, Stem Cell Technologies. Inc.). The remaining material was stained with antibodies for the markers Melan-A/Mart-1 (Dako) and HMB45 (Dako), CD133 (Miltenyi Biotec) and nestin (R&D System) and analysed by flow cytometry (BD FACSCalibur). Ten ml of blood were further processed and CD133, nestin, Melan-A/Mart-1 transcripts were quantified by Real Time RT-PCR (LightCycler, Roche Diagnostic). Results: CD45-depleted fractions in healthy controls were negative for melanoma markers. Melan-A/Mart-1 and/or HMB45 positive cells were detectable in 11 out of 12 melanoma patients. The absolute number of melanoma cells identified ranged from 6 to 176 per 10 ml of blood. Nestin expressing cells were more represented compared to CD133 expressing cells (median 27.4%, range 0.3% to 65.1% vs. median 9.3%, range 0.1% to 16.8%) within the melanoma fraction of cells positive for Melan-A/Mart-1 and/or HMB45. In one patient two different melanoma cell populations were detectable. The population of cells with lower expression of the melanoma markers showed at the same time higher expression of nestin and CD133 (5.9% vs. 1.3% and 10.2% vs. 6.7% respectively). Nestin results were in good accordance to the FACS data (nestin: r=0.55; CD133: r=0.23; Pearson test). Conclusions: The novel negative separation technique allows reliable isolation of melanoma cells from peripheral blood of patients with metastatic disease. A significant fraction of melanoma cells in peripheral blood bears a stem cell phenotype. No significant financial relationships to disclose.
Predictors of survival in patients with metastatic breast cancer (MBC) treated with high-dose chemotherapy and autologous stem cell transplantation (HD-ASCT)
Authors: Von Drygalski A. , Tran T. B. , Messer K. , Pu M. , Corringham S. , Nelson C. , Ball E. D. , Ball E. D. 
Published: August 2020

e22086 Background: Individualized care in MBC requires predictors of survival for tailored treatment. Although HD-ASCT has not resulted in improved overall survival (OS), retrospective analyses may identify patients who benefited. We reviewed records of all patients (n=96) in the bone marrow transplant registry at UCSD treated with HD-ASCT for MBC between 1989 and 2000. Methods: Age, race, stage at diagnosis, histology, estrogen receptor (ER) and menopausal status, body mass index (BMI) in kg/m2, time to transplant and death, sites of metastasis, disease status prior to and after transplant, and days in hospital were extracted. Brookmeyer & Crowley's 95% confidence intervals, Cox models for predictors of a time-to-event variable and Schoenfeld tests for proportional hazard assumptions were applied. Results: Median OS was 5.6 ys (CI 4.1–7.4) after initial diagnosis and 1.7 ys (CI 1.36–2.07) after transplant. OS after HD-ASCT at 12 ys was 8.2% and, although not statistically significant, 18.5% in ER- and 2.6% in ER+ patients, respectively. Stratified by ER status, stage at diagnosis was an independent predictor of OS. Patients with stage I at diagnosis were at lowest risk of death when compared to stage II-IV patients with HRs of 2.7 (II vs I CI 1.4–5.2), 4.6 (III vs I CI 2.1–10) and 17 (IV vs I CI 6.1- 47.8). Death risk was increased with BMI ≥ 30 (HR 3.1; CI 1.8–5.4), infiltrating lobular carcinoma (HR 2.5; CI 1.1–5.38) and visceral metastasis (HR 2.3; CI 1.3–4.1). Conclusions: The study highlights that ∼10% of patients experience ≥10 ys survival with HD- ASCT. Obesity, late stage at diagnosis, lobular infiltrating histology and visceral metastasis were independent negative predictors of OS. Surprisingly, ER- patients tended to live longer. Although survival was influenced by various disease characteristics, obesity was the only significant patient derived factor. How obesity negatively affects outcomes - whether affecting cancer growth or through associated morbidity - needs further study. These data may be useful stratification tools for future trials employing HD-ASCT as treatment modality in MBC. No significant financial relationships to disclose.
18F-Fluorothymidine (FLT) as tumor response predictor for anti-epidermal growth factor receptor (EGFR) antibody in human lung cancer xenograft
Authors: Takeuchi S. , Zhao S. , Kuge Y. , Zhao Y. , Jin Y. , Hatano T. , Shimizu Y. , Kinoshita I. , Dosaka-Akita H. , Tamaki N. 
Published: August 2020

e22088 Background: Specific EGFR inhibition therapies are currently promising and advanced in the clinical setting. It is revealed patients with a colorectal cancer bearing mutated K-ras do not benefit from cetuximab, a chimeric mouse-human antibody to EGFR. However, appropriate evaluation of the therapeutic effects has not been clarified yet, because of the lack of reliable tumor response predictors. Measurement of tumor proliferative activity by PET using FLT may serve to assess early response of the therapy. The purpose of this study is to determine whether 18F- FLT is useful to evaluate very early response to cetuximab treatment in human lung cancer cell line xenograft. Methods: Human tumor xenograft model was established in male BALB/c athymic mice by human lung cancer cell line NCI-H1975, which expresses L858R/T790M-EGFR. These mice were assigned to two groups (n=5, each group): control and cetuximab-treated groups. Cetuximab was intra-peritoneally given 1.0mg/body once on the first day. Biodistribution of 3H-FLT (%Inject Dose/g/kg) were studied two days after the treatment. Immunohistochemical staining of EGFR, phosphorylated EGFR (pEGFR), and nuclear antigen Ki-67 as a tumor cell proliferation marker in tumor tissues were also studied to assess early effects of cetuximab. Results: FLT uptake was high in tumors compared with any other organs. Two days after the cetuximab therapy, the mean value of FLT uptake in tumor was significantly decreased to 55% of control value (0.38 ± 0.03 in control group vs. 0.21 ± 0.07 in cetuximab-treated group; p < 0.05). There were no statistical differences in the tumor volume and body weight of mice between each group. High expression of EGFR and Ki-67 were observed in this xenograft. As compared with control, Ki-67 expression was remarkably decreased in cetuximab-treated group, while there were no definite changes in expression of EGFR and pEGFR. Conclusions: In this animal model, FLT uptake and Ki-67 expression were decreased in cetuximab- treated group just two days after the treatment start. These results indicate FLT-PET can be a useful predictor to evaluate very early response to molecular targeted medicine such as cetuximab before significant change of tumor size. No significant financial relationships to disclose.
Does hormone receptor (HR) positivity affect the prognosis in breast cancers with human epidermal growth factor receptor 2 (HER2) overexpression?
Authors: Lee Y. , Sohn J. , Park B. , Chung H. , Suh C. , Kim S. , Koo J. , Kim J. , Choi H. , Kim Y. 
Published: August 2020

e22091 Background: Biologically, there is an unclear issue about the role of HR positivity in HER2 positive breast cancer. These HER2(+)/ HR(+) pts were grouped into luminal B type apart from HER2(+)/ HR(-) pts in molecular profiling. However, from the clinical point of view, these pts have been categorized and been treated as either the only HER2(+) disease regardless of HR status or vice versa. Thus, we investigated the impact of HR status on clinical outcomes in HER2-overexpressed breast cancers. Methods: We retrospectively reviewed medical charts of HER2-positive breast cancer pts who underwent curative surgical resection from 1996 to 2001 in the Severance hospital, Korea. Demographic comparisons were performed by Chi-square tests. Tumor size, nodal stage, TNM stage, HR status, and adjuvant tamoxifen use were included in the Cox proportional hazards model. Results: Among the total 174 HER2-positive pts, HR (n=93) was positive in 53.5% (n = 93) and HR-positive tumors were more likely to be premenopausal (73% v 52%; P=0.01) and well- differentiated (grade 1or 2; 77% v 62%; P=0.04). There were no significant differences according to HR status in terms of tumor size, nodal stage, TNM stage, operation methods, and chemotherapy regimen. In these HER2-positive pts, the 5-year disease free survival (DFS) was longer in HR(+) pts than in HR(-) pts (DFS; 82.9% v 61.5%; P= 0.01). In a subset analysis, the 5-year DFS of HER2(+)/ER(+) pts without adjuvant tamoxifen (n=26) was not different from that of HER2(+)/ ER(-) pts (DFS; 57.7% v 61.5%; P= 0.32). However, the 5-year DFS of HER2(+)/ ER(+) pts with adjuvant tamoxifen was significantly prolonged compared with that of HER2(+)/ ER(-) pts (DFS; 91.5% v 61.5%; P< 0.001). In a multivariate analysis of DFS, tumor size and adjuvant tamoxifen use significantly affected DFS with an adjusted hazard ratio of 2.56 (95% CI, 1.2–4.9; P= 0.01) and 6.58 (95% CI, 2.8–20.3; P< 0.001), respectively. Conclusions: In an analysis of HER2-overexpressed breast cancer, the presence of HR itself did not affect the prognosis. However, most of the survival benefit seems to be driven from adjuvant tamoxifen therapy not the HR status itself. No significant financial relationships to disclose.
The role of EGFR MAbs C225 in breast cancer stem cells
Authors: Shi Y. , Zhang F. , Cao M. , Zhang Y. , Xu Y. , Huang M. , Wang H. , Ma Y. 
Published: August 2020

e22093 Background: The epithermal growth factor receptor(EGFR) signaling pathways has been implicated in self-renewal of breast cancer stem cells. It has been proved that the EGFR tyrosine kinase inhibitor-gefitinib significantly decreased the mammosphere- forming efficiency(MFE) in ductal carcinoma in situ (DCIS) derived cancer cells. We investigated the response of breast cancer stem cells in breast cancer cell line MCF-7 to EGFR MAbs C225(cetuximab). Methods: Cells were used for nonadherent (i.e.,mammosphere) culture and were divided into four groups according to the different culture medium whether or not including exogenous EGF and MAbs C225. MFE was calculated as the number of mammospheres (≥60um) and was expressed as a percentage. We Compared the percentage of CD44+/CD24- cells from the four groups, and measured the levels of the Wnt and Notch-3 gene expression of the four groups. Results: There was no significant difference of MFE between groups. In the presence of exogenous EGF, MAbs C225 significantly decreased the MFE and the percentage of CD44+/CD24- cells of mammospheres compared with exogenous EGF alone(0.7% versus 1.5%, P < 0.001 and 4% versus 11%P < 0.001;respectively). In the absence of exogenous EGF, MAbs C225 aslo statistically decreased the MFE and the percentage of CD44+/CD24- cells of mammospheres compared with that of lacking exogenous EGF and MAbs C225(0.53% versus 1.3%,P = 0.003 and 9% versus 3%, P < 0.01;respectively). And we observed that the Notch-3 gene expression was compensativly increased in the two groups of addition of MAb C225. No significant differences of Wnt gene expression were observed between groups. Conclusions: EGFR MAbs C225 could disrupt mammoshpere formation and decrease the percentage of CD44+/CD24- cells of mammoshpere cells. The expression of Notch-3 is increased with the EGFR antagonistics in breast cancer stem cells. No significant financial relationships to disclose.
Molecular analysis of tamoxifen-resistant pathways in breast cancer cell-lines and breast cancer tissues
Authors: Yeoh C. , Chelala C. 
Published: August 2020

e22094 Background: Oestrogen receptor positive (ER+) breast cancer (BC) is heterogeneous with regard to their clinical behaviour and response. The ER and Progesterone receptors (PgR) are currently the best predictors of response to the anti-oestrogen agent tamoxifen, yet up to 30–40% of ER+ breast cancers will relapse despite tamoxifen treatment. New prognostic biomarkers and further biological understanding of tamoxifen resistance is required. There has been an explosion of greater understanding since the arrival of new cutting-edge genomic profiling technology. Methods: RNA from the BC cell lines and frozen tissue was extracted and qualified using Qiagen Mini Kit and BioAgilent respectivelly. Probe preparation and scanning of the chips were carried out according to Affymetrix protocol. The normalisation and analysis of data (Limma) was performed using Bioconductor software. RT- PCR probes were purchased from Applied Biosystem and performed on ABI7500 instrument. The IHC-P was done using Ventana. TMA prepared with TMABooster, Alphelys (Plaisir, France). Results: Gene expression changes were studied in the TR breast cell lines MCF7 (tenovus), T47D and ZR by using the Affymetrix Hu133 plus 2.0 platform. Common genes of significant expression from the 3 TR cell lines were validated by real-time PCR (RT-PCR) and immuno-histochemistry (IHC-P) staining. 7 (MAGEA2, AKR1C3, VGLL1, THRAP5, GREB1, PDZK1, MYBL1, HnRNPA2B1, PGR) out of the 9 chosen genes were successfully validated. Proliferation studies on MageA2 overexpressing clones were performed. Stable overexpression of MageA2 in T47D and MCF-7 confers a proliferation advantage in tamoxifen-containing media as shown in proliferation and cell cycle analysis. MageA2 has been shown to alter the p53 pathway. The expression of KAP1, which interacts with p53 and inactivates it, was enhanced by MageA2 overexpression. In addition, overexpression of MageA2 is seen to increases histone deacetylase (HDAC3), and p53 expression as seen by immunoblotting. Validation of MageA2 on TMA of 100 patients is underway. Conclusions: MageA2 is a possible selective molecular target for the reversal of tamoxifen resistance. No significant financial relationships to disclose.
Triple-negative breast cancer in Hong Kong Chinese patients
Authors: Tsang J. , Lai T. L. , Lau D. H. , Au G. K. , Chua D. T. 
Published: August 2020

e22127 Background: There is increasing data showing that breast cancer is a heterogeneous disease which should be assessed separately in different populations, as it differs substantially between Chinese and Caucasian women. Triple-negative breast tumours which are negative for ER, PR and HER-2 neu receptors are associated with younger age at presentation, tumour of higher grade with larger size and a poorer prognosis. There is recent suggestion that the prognostic outlook of Chinese triple-negative breast cancers might be somewhat different from those in the Western population, but few studies have attempted to understand the role of ethnic factor in the triple-negative entity. Methods: We conducted a preliminary retrospective comparison of 170 Hong Kong Chinese primary breast cancer patients seen as new cases during January 2004 and December 2004 in a teaching hospital. Clinico-pathological features of triple-negative tumours were compared to their non-triple-negative counterpart. Results: Triple negative breast cancer accounted for 12.4% of all breast cancer patients seen in the year of 2004 (n = 21). It is associated with more cancers with grade 3 tumour (68.4% vs 36.8%; p = 0.02) but there was no statistically difference between the age of presentation, tumour size, extensive intraductal component, lymph node status and rate of local relapse or metastasis after adjuvant therapy. Subset analysis further revealed that when triple negative breast cancer patients (n = 21) were compared to the HER-2 positive patients (n = 40) in the studied population, HER-2 positive patients were still associated with higher proportion of node positive disease (57.5% vs 30.0%; p = 0.04). Disease free survival and overall survival were not studied due to limited follow-up time. Conclusions: Our preliminary findings suggested that Hong Kong Chinese triple-negative breast cancers are associated with a more favourable outlook and might behave differently when compared to their Western counterpart. Further large-scale study of the ethnic factor with long-term follow-up is warranted. No significant financial relationships to disclose.
Role of E1B protein-dificient oncolytic virus in breast cancer stem cells
Authors: Zhang F. , Ma Y. , Xu Y. , Huang M. , Song C. , Lin S. , Zhang Y. 
Published: August 2020

e22131 Background: Cancer stem cells have been indicated in the initiation of tumors and are even found to be responsible for relapses after apparently curative therapies have been undertaken. In breast cancer, they may reside in the CD44+CD24−/low population. Oncolytic adenoviruses enter cells through infection and can kill both proliferating and quiescent cells. We investigated the role of E1B protein- dificient oncolytic adenovirus in breast cancer stem cells. Methods: MCF-7 cells were infected by E1B protein-dificient oncolytic adenovirus as infected group (MOI=100) and cultured routinely as control group simultaneously. The proportion of CD44+CD24- cells was assessed by flow cytometry (FCM) in two groups respectively. Meanwhile, mammosphere culture was done in two groups' cells to observe the size and number of mammospheres, calculate the mammosphere- forming efficiency (MFE). The proportion of CD44+CD24- cells in two groups' mammospheres was assessed by FCM. Results: The percentages of CD24-,CD44+, CD44+CD24- in the infected gruop were 43.9%, 63.26%, 22.19%, respectively. While in the control group, the percentages were 6.74%, 88.30%, 2.30%. In the infected group, the time of mammosphere's formation was earlier, the volumes of mammospheres were bigger and the MFE was higher than the control group (1.26%:0.9%). In two mammospheres' groups, the proportion of CD44+CD24- cells in experiment group and control group was 38.08% and 23.35%, respectively. Conclusions: E1B protein-dificient oncolytic adenovirus can kill MCF-7 cells in short time, mainly breast cancer differentiated cells. It maybe promote the growth of the breast cancer stem cells. It maybe accelerate the speed of self-renewed and differentiation of the breast cancer stem cells. No significant financial relationships to disclose.