bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of a combination of antiangiogenic and antilymphangiogenic therapies on a death receptor-5 mediated antitumor immunotherapy in mice.
Authors: Tsukita Yoko , Okazaki Tatsuma , Komatsu Riyo , Nihei Mayumi , Kobayashi Makoto , Sato Yasufumi , Yagita Hideo , Ichinose Masakazu 
Published: September 2018

e23001 Background: In angiogenesis, VEGF-A and its receptor VEGFR-2, and in lymphangiogenesis, VEGF-C and VEGFR-3, play central roles. An anti-tumor immunotherapy using an agonistic antibody (Ab) to death receptor (DR)-5, the apoptosis inducing receptor, exhibits antitumor effects. VEGF-A or -C inhibition augmented the effects of immunotherapy. However, little is known about the effects of combining anti-angiogenesis, anti-lymphangiogenesis, and immunotherapy. Here, we combined sunitinib, which mainly inhibits VEGFR1-3 and PDGFRs, as anti-angiogenic and anti-lymphangiogenic therapies, with anti-DR-5 Ab as an immunotherapy in mice. Methods: Mice were injected with breast carcinoma 4T1 cells into the footpad, and treated with sunitinib and/or anti-DR-5 Ab. On day 14, mice were anesthetized, and the popliteal lymph nodes and tumors were isolated for immunohistochemistry, flow cytometry, hypoxia, and lymphatic flow analysis. To evaluate the tumor growth rate, mice were injected with 4T1 cells into the hindflank, treated with the combination or monotherapy and sacrificed on day 21. CD4+ and/or CD8+T cells were depleted by anti-CD4 and/or anti-CD8 Ab, respectively. Results: Sunitinib decreased endothelial cells, pericytes, and hypoxic area densities in the tumors suggesting blood vessel normalization. Sunitinib also decreased the amount of evans blue injected into tumors and hyaluronic acid in tumors, suggesting improved lymphatic flow. In draining lymph nodes, sunitinib and anti-DR-5 Ab combination increased the numbers of activated dendritic cells and CD8+ T cells than controls. In tumors, the combination increased the numbers of infiltrating effector CD4+FOXP3- and CD8+ T cells compared to controls. Sunitinib and anti-DR-5 Ab combination augmented the decrease in the tumor growth rate compared to each mono-therapy. CD4+ and/or CD8+ T cell depletion showed anti-tumor effects of the combination were partially CD8+T cell-dependent. Conclusions: Blood vessel normalization and improved lymphatic flow in tumors may improve circulation between the tumors and draining lymph nodes, which has a potential to augment the effects of immunotherapy.
Analysis of multidrug resistant transporter expression in tumor blood vessels of urothelial carcinoma during chemotherapy.
Authors: Kikuchi Hiroshi , Maishi Nako , Akiyama Kosuke , Morimoto Masahiro , Yanagiya Misa , Miyajima Naoto , Tsuchiya Kunihiko , Maruyama Satoshi , Abe Takashige , Hida Yasuhiro , Harabayashi Toru , Ameda Kaname , Matsumoto Ryuji , Kashiwagi Akira , Matsuno Yoshihiro , Shinohara Nobuo , Hida Kyoko ,   Hokkaido University Oncology Group
Published: September 2018

e23004 Background: ABCB1, a multidrug transporter, is encoded by multidrug resistance (MDR) 1 gene and plays a major role in drug resistance. Recently, we have reported that not only tumor cells, tumor vascular endothelial cells (TECs) also confer cancer drug resistance. We isolated TECs and found that they have various abnormalities such as aneuploidy or stemness characteristics. TECs showed resistance to Paclitaxel (PTX) with ABCB1 up-regulation. Furthermore, their resistance to PTX was abrogated by ABCB1 inhibition of TECs in vivo. In clinical urology, gemcitabine / cisplatin (GC) is standard 1st line chemotherapy for metastatic urothelial carcinoma (mUC). PTX is often selected in 2nd line chemotherapy for GC resistant cases, however the therapeutic outcomes are limited. We hypothesized that ABCB1 inhibitor/PTX combination would be more effective strategy for mUC, if TEC ABCB1 expression is upregulated. In this study, we investigated ABCB1 expression in tumor blood vessels of UC during 1st line chemotherapy. Methods: Paraffin-embedded samples were corrected from 50 patients who were performed tumor resection before and after 1st line chemotherapy. ABCB1 expressions were analyzed by immunohistochemical staining and the ratio of ABCB1 positive (+) vessels in total vessels were quantified. In vitro assays were performed to address how endothelial cells (ECs) are affected by change of tumor microenvironment during chemotherapy. Results: The ABCB1 (+) vessels were 0.00 – 16.80% (Median 0.00%) and 0.00-58.94% (Median 4.49%), before and after chemotherapy, respectively. In 32 of 50 cases (64%), the ratio of ABCB1 (+) vessels increased after 1st line chemotherapy. Additionally, gemcitabine induced MDR1 mRNA expression in ECs via NF-kB activation. Furthermore, gemcitabine and cisplatin induced IL-8 secretion from tumor cells, and MDR1 mRNA expression level was elevated in ECs. Conclusions: It was suggested that conventional chemotherapy may cause inflammatory change in tumor tissues, which causes in ABC transporter induction in tumor blood vessels. It was suggested that inhibition of ABC transporter in TECs is one of important new strategy to overcome cancer drug resistance.
Can 18F-alfatide micro-PET predict the radiotherapy response in Lewis lung carcinoma tumor-bearing C57BL/6 mice?
Authors: Wei Yuchun , Liu Ning , Huang Yong , Hu Xudong , Yuan Shuanghu 
Published: September 2018

e23011 Background: Integrin αvβ3 plays a major role in angiogenesis, tumor growth and progression. A new tracer, 18F-AL-NOTA-PRGD2, denoted as 18F-Alfatide, has been developed for positron emission tomography (PET) imaging of integrin αvβ3. This study aimed to explore the predictive role of integrin αvβ3 imaging with 18F-Alfatide PET in non-small cell lung cancer (NSCLC) for radiation and also to assess changes in tumor uptake during the course of radiotherapy. Methods: A total of 40 mice with lewis lung carcinoma (LLC) xenografts were divided into 4 groups (Ⅰ, Ⅱ, Ⅲ and Ⅳ) randomly. The Ⅰ underwent 18F-Alfatide Micro-PET scanning before and after radiotherapy, and so did the Ⅱ except for radiotherapy. The xenografts of Ⅲ were removed at the time before radiotherapy, while the Ⅳ’s were removed after radiotherapy. Tumor and muscle uptakes before and after radiotherapy of Ⅰ(SUVrad-1, SUVrad-2, SUVM-1, SUVM-2) and Ⅱ(SUVblank-1, SUVblank-2, SUVm-1, SUVm-2) depict 18F-Alfatide Micro-PET uptake. The expression of integrin αvβ3 in Ⅲ and Ⅳ xenografts was analyzed by western blotting. Results: The changes of ΔSUV1 ( SUVrad-2/SUVM-2―SUVrad-1/SUVM-1) was significantly lower than ΔSUV2 (SUVblank-2/ SUVm-2―SUVblank-1/ SUVm-1 ) (2.65±1.72 versus 5.16±2.50, P = 0.019. Overall survival (OS) of Ⅰ(56.70±25.34 day) is significantly longer than Ⅱ(35.00±5.27 day), P = 0.002. Significant negative correlation was found between ΔSUV and OS (R = 0.640, P = 0.002). Significant higher αvβ3 was observed in Ⅲ(0.52±0.09) than Ⅳ(0.41±0.07), P = 0.007. Conclusions: Decrease of ΔSUV and integrin αvβ3 during the course of radiotherapy may predict improved response in LLC xenografts and affirm the predictive role of 18F-Alfatide for radiation in NSCLC.
The novel long non-coding RNA PANCR: A p53 activator and potential breast cancer biomarkers.
Authors: Cao Yu , Cao Deliang , Ling Hongyan 
Published: September 2018

e23016 Background: Long non-coding RNAs (LncRNAs) may serve as a biomarker and potential therapeutic target of cancers. Chromosome 16q22.1 is frequently deleted in breast cancer and may contribute to breast carcinogenesis by inactivation of tumor suppressor genes. This study characterized a new LncRNA tumor suppressor in this region, named p53 activating non-coding RNA (PANCR). This LncRNA consists of 1.5kb in length. Methods: Quantitative real-time PCR was used for examine the PANCR expression in breast cancer tissues. RNA-pull down and RNA-Immunopreicitation were used to analyze PANCR targeted protein. Results: Our data showed that PANCR was downregulated in breast cancer cell lines and tissues. In the breast cancer cell lines, PANCR expression appeared reversely correlated with cell malignancy, and in breast cancer tissues, PANCR was downregulated over 2 times in 31(62.0%) of 50 cases compared to adjacent normal breast tissues. In breast cancer cells MCF7 and immortalized human mammary epithelial cells MCF10A, ectopic expression of PANCR induced marked apoptosis, suppressing cell proliferation in culture and tumor growth in xenografts, but in contrast, shRNA–mediated silencing of PANCR promoted cell growth and proliferation. Mechanistic approaches revealed that in both MCF7 and MCF10A cell, PANCR activated p53 and upregulated pro-apoptotic proteins bid and bim and cell cycle inhibitors p21waf/cip1 and p27Kip1. We further found that the PANCR binds to and activates p53 by dissociating the p53-MDM2 complex. We further characterized the functional domain of PANCR that interacts with p53. Conclusions: The LncRNA PANCR located in the deleted Chromosome 16q22.1 region is a novel intracellular p53 activator and tumor suppressor, which may be used as a target for cancer therapy through mimicking its binding domain and activation of p53.
Immunomonitoring in patients with stage II and III colon cancer.
Authors: Vocka Michal , Spacek Jan , Netikova Irena , Zavadova Eva , Němejcová Kristýna , Konopasek Bohuslav , Dundr Pavel , Geppert Carol-Immanuel , Fucikova Terezie , Hartmann Arndt , Petruzelka Lubos B. 
Published: September 2018

e23038 Background: Colorectal cancer (CRC) is the second leading cause of cancer death worldwide. Immunoscore has been shown to be a very powerful prognositc indicator in patients with clinically localized colorectal cancer, with no detectable tumour spread to lymph nodes or distant organs. These patients are usually treated with only surgical removal of the tumour; however, approximately 25% of these patients will have recurrence of their disease, indicating that occult metastases were already present at the time of curative surgery. VEGF is the factor responsible for neoangiogenesis and it is being considered as a possible prognostic marker of disease progression. Transforming growth factor-beta (TGF-beta) is also neoangiogenic and a highly immunosuppressive factor. The purpose of this study was to monitor the immune response in patients with stage II and III CRC, with a focus on cellular as well as humoral immunity. TGF-beta and VEGF levels were followed. Methods: 25 patients with stage II and III colon cancer included in the research project received routine cancer teratment. Basic parameters - histological type and grade - were established at baseline. Patients were evaluated by a clinical immunooncolgist to exclude any immune disorders of allergic or autoimmune origin. TGF-beta and VEGF were measured using ELISA, and anti-tumour cellular immunity (CD4, CD8, T-reg, B cells ) were measured via flow cytometry. Results: In patients with stage II and III colorectal cancer, predominantly a depression in cellular immunity was seen. Plasma levels of immunglobulins were also reduced, particularly the IgG4 subtype. Most patients showed some clinical symptoms of immunodeficiency, such as frequent respiratory tract infections and/or herpetic infections. TGF-beta and VEGF plasma levels were increased. Conclusions: The correlation of these neoangiogenic and immunosuppressive factors, as well as the state of anticancer immunity with the clinical outcome of the patients, could help in the future as a prognostic marker and contribute to the selection of targeted immune therapy in patients with colorectal cancer. Dedication: Ggrant of Czech Ministery of Health, 15-28188A, League against cancer and PRVOUK-P-27/LF1/1
Dynamic process of circulating tumour DNA (ctDNA) profiling as a predictor of efficacy of pemetrexed combined platinum chemotherapy in advanced lung adenocarcinoma.
Authors: Han Ying , Han Xiaohong , Wang Zhe , Luo Wen , Zheng Wenyuan , Dai Wumin , Yu Huang , Yao Jiarui , Zhang Zhishang , Wu Di , Shi Yuankai 
Published: September 2018

e23048 Background: Pemetrexed combined platinum is recommended as first line chemotherapy regimen for non-squamous non-small cell lung cancer. However, the efficacy is variable for individuals. Methods: We conducted a prospective study with 52 consecutive patients enrolled who were histologically confirmed advanced or metastatic lung adenocarcinoma, of whom, 32 had their plasma samples collected both baseline and follow-up when efficacy evaluation. We used a next-generation sequencing to assess the dynamic change of ctDNA across 87 plasma samples. Results: 34 blood samples from 61.5% of patients demonstrated smoking status is an infavorable factor to drug efficacy ( p= 0.02, fisher’s exact test, two-sided). Simultaneously, we found positive correlation between decreased mutation burden and chemotherapy sensitivity (the mean mutation rates of pre chemotherapy, PD, PR, SD and SDA were 4.5, 6.3, 2.1, 1.6 and 2.4, respectively, p= 0.004, one way ANOVA test, two-sided).The mean mutation rates with T > G substitution mutation of pre and post chemotherapy were 1.5 and 0.6 ( p= 0.014, Mann-Whitney U test, two-sided). 34.4% (11/32) individuals harbor those mutation pre chemotherapy, 6 out of them were characterized by DNA repair signatures, of which 83.3% (5/6) cases were positively related to platinum sensitivity. In addition, T > G substitution was still detected from 5 patients at cycle 2, 60% (3/5) individuals were characterized by DNA repair signatures and 2 out of them had better platinum-based chemotherapy response.Neoplastic cells carrying a specific set of somatic mutations were sensitive to platinum treatment, including USP9X (pre vs post chemotherapy: 10.3% vs 0%), ZRSR2 (pre vs post chemotherapy: 6.9% vs 0%) and PIK3C2G (pre vs post chemotherapy: 10.3% vs 0%). Nonetheless, persistence of gene ROBO2 (pre vs post chemotherapy: 10.3% vs 4.6%) may be correlated with tumorigenesis. Conclusions: Dynamic monitoring of ctDNA mutation status may be contributed to advanced or metastatic lung adenocarcinoma patients stratification and prediction for pemetrexed combined platinum as first line chemotherapy regimen.
Enumeration of circulating tumor cells using a non–EpCAM-dependent device.
Authors: Amato Robert J. , Brobey Reynolds , Wang Yue , Murphy Kent , Smith Jeff 
Published: September 2018

e23064 Background: One component of the metastatic process is the circulation of tumor cells. Identification of these circulating tumor cells (CTCs) provides important insights into cancer biology. There is enormous heterogeneity among CTCs, necessitating capture of both epithelial and non-epithelial subtypes. Detection and monitoring of CTCs will provide a non-invasive biomarker for the detection of early progression and assessing therapeutic response. Methods: The AxonDx nCyte™ System is designed to detect rare circulating cells from peripheral blood without enrichment or selection. An epi-fluorescence microscope scans pathology glass slides on which a sample has been applied, and high-resolution images of potential CTCs are captured at four emission wavelengths. The proprietary Cancer Cell Detection Cocktail is designed to detect CTCs from numerous solid tumors, including various stages (e.g., mesenchymal-epithelial transformation) and stem cells. The software distinguishes cancer-derived material (nucleus+/cytokeratin+/WBC marker-/ > 4 µm); from leukocytes (nucleus+/cytokeratin-/WBC marker+). Vimentin is used to distinguish epithelial from mesenchymal CTCs. We obtained informed consent from 32 genitourinary cancer patients to participate in this IRB-approved study. Red blood cells from 6 mL of whole blood were lysed and removed from 32 patient samples. Nucleated cells from the blood sample were fixed, permeabilized, and stained with AxonDx’s Cancer Cell Detection Cocktail. The prepared cells were dispensed onto two microscope slides, cover-slipped, and analyzed. Results: We detected CTCs in 32 of 32 samples (100%), in a range of 0.6–64.3 CTCs/mL. Number of CTCs correlated with metastatic tumor burden. Conclusions: The AxonDx nCYTE™ system can reliably detect CTCs from genitourinary patient samples. The AxonDx nCyte™ System has also successfully detected CTCs in other tumor types, e.g., breast, lung, CRC, and HCC. Because the system is not EpCAM-dependent, it captures the heterogeneous population of CTCs. Prospective longitudinal studies are ongoing to determine trends in CTC counts.
Effects of hydrogen gas on exhausted CD8+ T cells (PD-1-positive, terminally differentiated CD8+ T cells).
Authors: Akagi Junji , Baba Hideo 
Published: September 2018

e23071 Background: CD8+ T cells progress from an early-differentiated (early-CD8+ T cells) to a terminally differentiated state (terminal-CD8+ T cells). PD-1 expression is rapidly upregulated in antigen-activated T cells, and rapidly downregulated when the antigen is cleared. In the presence of persistent antigen stimulation, such as that in cancer patients, PD-1 expression is not downregulated and terminal-CD8+T cells become exhausted. Methods: Using flow cytometry, we investigated PD-1 expression in CD8+T cells in the peripheral blood of 37 patients with stage IV cancer, before and after they received hydrogen gas treatment. We analyzed associations between treatment and progression-free survival (PFS) using Cox and Kaplan-Meier survival analyses. Results: Treatment lead to partial response (PR) in 12 patients and stable disease (SD) in 14 patients, with response rates of 32.4% and 37.8%, respectively (with an overall clinical response rate of 70.3%), and reduced the number of PD-1-positive, terminal-CD8+ T cells in 9 of 12 patients with PR (75%) and in 8 of 14 patients with SD (57%). However, in 8 of 11 patients with progressive disease (PD) (72.7%), these cell counts increased. Multivariate analyses demonstrated that higher PD-1-positive, terminal-CD8+ T cell counts were associated with shorter PFS periods, both before (hazard ratio [HR] = 3.23; 95% confidence interval [CI], 1.475–7.070; p = 0.003) and after (HR = 2.75; 95% CI, 1.273–5.951; p = 0.01) treatment, although the hazard ratio was decreased by treatment. Additional analysis showed that patients with higher PD-1-positive, terminal-CD8+ T cell counts had shorter survival rates than those with lower counts, both before and after treatment (log rank test, p = 0.001 and p = 0.005, respectively). The 20-month survival rates for patients with higher PD-1-positive, terminal-CD8+T cell counts improved from 0% to 36.7% after treatment. Conclusions: These results suggest that PD-1-positive, terminal-CD8+ T cell counts are highly associated with PFS duration in patients with stage IV carcinomas, and hydrogen gas treatment contributes to clinical outcomes by reducing PD-1-positive, terminal-CD8+ T cell counts.
Role of the inflammasome of tumor cells in modulating the biology of Tumor Infiltrating T lymphocytes (TILs) in colorectal cancer.
Authors: Jarry Anne , Ouairy Adrien , Dansette Delphine , Deleine Cécile , Jouand Nicolas , Oger Romain , Podevin Juliette , Fourquier Pierre , Thibaudeau Emilie , Chetritt Jerome , Mosnier Jean-François , Gervois Nadine , Bossard Celine 
Published: September 2018

e23087 Background: In colorectal cancer (CRC), little is known about mechanisms by which tumor cells can influence the phenotype and biology of Tumor Infiltrating T lymphocytes (TILs) of the tumor microenvironment. One of these mechanisms could be the inflammasome, a molecular platform present in normal intestinal epithelial cells, whose effector protein, caspase-1, can rapidly mature IL18 and generate a mucosal Th1/Tc1 (IFNγ) response. However, the inflammasome status of tumor cells in CRC and its potential role on TILs are unknown yet. Methods: Prospective and retrospective cohort studies aimed to determine in CRC patients: the status of the inflammasome in tumor cells (IL18 immunostaining on tissue microarrays (TMA) and in situ detection of active caspase-1 on frozen sections) and the density of TILs (CD8+, T-bet+) in relation with i) the microsatellite stable (MSS) or unstable (MSI) status of CRC, and ii) the levels of cytokines (IL18, IFNγ) secreted in an ex vivo explant culture model of CRC. Finally, we assessed the effect of recombinant human IL18 (rhIL18) on the IFNγ response of isolated TILs. Results: TMA analysis of the retrospective cohort showed that IL18 was significantly expressed (in more than 50% of tumor cells) in 80% of CRC, especially in MSI CRC, and correlated with a high density of T-bet+ and CD8+ intraepithelial TILs (IEL-TILs). Active caspase-1 was detected in tumor cells in 60% of CRC. In the prospective cohort, the presence of active caspase-1 in tumor cells was associated with high levels of mature IL18 secreted in explant cultures, with high density of T-bet+ TILs and with IFNγ release in most cases. In addition, isolated TILs expressing IL18 receptors (IL18Rα), cultured with rhIL18, were able to secrete IFNγ either unstimulated or stimulated with OKT3. Conclusions: The inflammasome of tumor cells, when maintained and active, can contribute to a Th1/Tc1 antitumor immune response elicited by TILs, that can modulate tumor growth. The inflammasome of tumor cells can thus be considered as a potential new therapeutic target to strengthen the antitumor immune response in CRC, in association with other immunotherapies.
A new method for gene expression analysis of pure lymphocytes recovered from heterogeneous fixed mouse tissue.
Authors: Chow Jennifer , Da Silva Ana Paula Galvão , Medoro Gianni , Manaresi Nicolò , Lira Paul David , Bischoff Farideh Z. 
Published: September 2018

e23089 Background: Tumor infiltrating lymphocytes (TILs) are biomarkers that play a critical role in cancer diseases, including differential diagnosis, determination of prognosis, prediction of response to treatment, and evaluation of disease progression. Gene expression analysis in TILs derived from fresh tissue may not accurately depict the gene profile of the tissue microenvironment as it can change aggressively during lymphocyte isolation and RNA extraction. In addition, tissue sample size can limit the isolation of TILs with current technologies. In this study, we demonstrate the use of the DEPArray™platform to isolate pure populations of lymphocytes from a fixed mouse tissue for RNA analysi. Methods: Mouse splenocytes were activated in vitro with anti-CD3 and -CD28 for 72hs. Cells were harvested, fixed with 2% paraformaldehyde (PFA) for 20 min at RT, and stained for either CD4 or CD8 expression. Gene expression analysis of CD45, ADORA2A, GLS and GAPDH was performed in CD4+ and CD8+ DEPArray™sorted cells using the TaqMan PreAmp Cells-to-Ct kit. Results: The table below summarizes the Ct values for CD45, ADORA2A, GLS and GAPDH expression in 300 fixed unsorted control and DEPArray™sorted lymphocytes. Conclusions: We have demonstrated the feasibility of gene expression analysis on pure populations of CD4+ and CD8+ cells isolated from a fixed tissue using the DEPArray™ platform. The advantage of this approach is the DEPArray’s ability to identify and isolate subpopulations of cells from complex heterogeneous samples and/or specimens that are limited by size or content. This methodology will be applied for isolation of TILs in syngeneic and xenograft models of cancers for downstream RNA applications. [Table: see text]