bluuebunny/crossref_metadata_2025_split huggingface dataset:
In vivo real-time imaging of tumor microcirculatory alterations in colorectal liver metastatic xenografts by chemotherapy using multiphoton microscopy.
Authors: Tanaka Koji , Okigami Masato , Toiyama Yuji , Morimoto Yuhki , Matsushita Kohei , Kawamura Mikio , Hashimoto Kiyoshi , Saigusa Susumu , Okugawa Yoshinaga , Hiro Junichiro , Inoue Yasuhiro , Uchida Keiichi , Araki Toshimitsu , Mohri Yasuhiko , Mizoguchi Akira , Kusunoki Masato 
Published: March 2020

e21053 Background: Tumor endothelium is a distinct target for anticancer treatments of metastatic colorectal cancer. Various chemotherapeutics themselves have anti-angiogenic effects on tumor microenvironment by targeting proliferative endothelial cells during tumor angiogenesis. In this study, we imaged the dynamics of tumor microcirculation of colorectal liver metastasis in living mice in vivo real-time using two-photon laser scanning microscopy (TPLSM), and evaluated the microcirculatory alterations in tumor microenvironment by chemotherapy. Methods: Red fluorescent protein expressing human colorectal cancer cell line (HT29) was inoculated to the spleen of green fluorescent protein expressing nude mice. 5-fluorouracil or irinotecan was administered three times a week for more than three weeks for metronomic scheduling. Intravital TPLSM was performed at multiple time points for time-series imaging of liver metastatic xenografts in the same mice. The alterations in tumor microcirculation during chemotherapy was evaluated by measuring blood flows at tumor vessels of colorectal liver metastasis and hepatic sinusoids of adjacent normal liver. Results: At the first TPLSM imaging, the blood flow, as determined from the movement of platelets, was heterogeneous and non-directional in the tumor vessels of liver metastatic xenografts. The blood flow was relatively slower in tumor vessels than normal hepatic sinusoids. At the second TPLSM imaging after chemotherapy, platelet aggregation was observed in tumor vessels of the same mice. Aggregated platelets were frequently adhered to the tumor endothelium, suggesting tumor vessel damage or intratumoral coagulation abnormality by chemotherapy. There was no difference in chemotherapeutics with regard to these findings. Conclusions: Intravital TPLSM imaging of tumor microcirculation at metastatic tumor xenografts is a useful tool to evaluate anti-angiogenic drugs in preclinical models.
Expressions of KLF2 and chemokine receptor CCR7 in pulmonary adenocarcinoma.
Authors: Itakura Meiji , Shingoji Masato , Iizasa Toshihiko 
Published: March 2020

e21054 Background: Chemokines and chemokine receptors not only have the powerful ability in cancer metastasis and tumorigenesis, but also act as anti-tumorgenic ability. Lung Krueppel-like factor (LKLF, KLF2) is a member of the family of the Krueppel-like factors (KLFs). KLF2 was initially described as a lung-specific transcription factor. KLF2 is reported to regulate some malignant cells. We examined and evaluated the effect of KLF2 on pulmonary adenocarcinoma and the relationship of their mRNA expression with CCR7, EGFR and p53 genetical mutations in pulmonary adenocarcinoma. Methods: 120 patients of stage I to IV with pulmonary adenocarcinoma were included in this retrospective analysis. The expression of CCR7 and KLF2 mRNA expression in surgically resected pulmonary adenocarcinoma specimens were examined and evaluated the relation to prognosis, the effect of EGFR and p53 genetical mutations. Results: High mRNA expression of KLF2 in lung cancer patients indicated significantly good prognosis than the groups of low expressions (p= 0.0066, HR= 2.008, 95% CI of ratio 1.215 to 3.319). The expression of KLF2 mRNA had relationships with CCR7 mRNA expression in pulmonary adenocarcinoma. Moreover the mRNA expression of KLF2 in pulmonary adenocarcinoma specimens was influenced by the mutation of p53 mutation in lung cancer specimens. There was no significant difference within KLF2 mRNA expression quantities of EGFR mutation positive and negative group. Conclusions: We propose KLF2 as clinical good prognostic factors and that KLF2 has strong relation with CCR7 and p53 genetical mutation mRNA expression in pulmonary adenocarcinoma.
ApoStream, an antibody-independent platform, compared to CellSearch for enumeration of circulating tumor cells (CTCs) in patients with metastatic prostate cancer.
Authors: Poklepovic Andrew Stewart , Wan Wen , Wu Weiguo , Woo Jacky , Melnikova Vladislava , Jouravleva Elena , Anderes Kenna Lynn , Davis Darren 
Published: March 2020

e21058 Background: Development of CTC detection platforms is a rapidly advancing field. The CellSearch technique relies on cell surface expression of EpCAM to select for rare tumor cells in whole blood specimens. Consequently, the use of EpCAM-based enrichment platforms to recover CTCs is limited to EpCAM expressing cells and is poorly suited for recovery of EpCAM negative cells like those having undergone epithelial mesenchymal transition (EMT). ApoStream is an antibody independent enrichment platform which utilizes the principle of continuous flow dielectrophoresis field-flow fractionation (DEP-FFF) to isolate and enrich for CTCs. A head to head comparison of CellSearch to ApoStream for recovery of CTCs in patients with metastatic prostate cancer was performed. Methods: Two 7.5 mL bloodsamples were collected at a single time point for each patient with stage IV prostate cancer. One sample was analyzed by CellSearch CTC enumeration kit, and one sample was analyzed by ApoStream. CTCs recovered by both devices were immunophenotyped using antibodies against cytokeratin (CK), CD45 and DAPI. CTCs were defined as CK+/CD45-/DAPI+ intact cells. CTCs recovered by ApoStream were further analyzed by quantitative laser scanning cytometry (LSC). A paired t-test was used to compare the cell counts in the two devices. Results: The ApoStream CTC enrichment platform isolated a greater number of CTCs from blood of eight patients compared to CellSearch (p= 0.0027). All cell counts obtained by the ApoStream technique were higher than CellSearch, and all patients had detectable CTCs by ApoStream, while only 75% of patients had detectable CTCs with CellSearch. Conclusions: The ApoStream platform is differentiated from EpCAM dependent platforms and is well suited for detection and recovery of CTCs in advanced stage disease where tumor cell heterogeneity is common and expression of EpCAM may be low or lost. [Table: see text]
The novel receptor targeted (CD206) 99mTc-labeled tilmanocept versus the currently employed Tc99m-sulfur colloid in intraoperative lymphatic mapping (ILM) on key performance metrics breast cancer.
Authors: Cope Frederick Oliver , Metz Wendy , Potter Beth , Abbruzzese Bonnie , Blue Michael , Shuping Jo , Christman Lori , King Dennis 
Published: March 2020

e21066 Background: ILM is a key diagnostic component for solid tumor excision. ILM utilizes the injection of a color agent and/or a radiopharmaceutical agent (RPA) which may be tracked intraoperatively w/ a γ-probe. ILM is designed to aid the surgeon in tracing lymph nodes that may have a connection to the tumor bed. ILM in the U.S. employs a non-standard RPA, S-colloid (TcSC). This agent was recently approved by the FDA for breast cancer ILM using a literature review. In contrast, CD206-targeted Tc99m-tilmanocept (TCP) was studied in 2 prospective Ph-3 trials. These trials compared TCP to blue dye, the same dye utilized in the FDA review of TcSC. Methods: For a statistical review and contrast of TCP vs TcSC, we used a meta-analysis and pooled analysis based on the FDA-review of TcSC. We focused on the functional endpoints: 1] Localization rate (% of pts w/ a found hot node) and 2] degree of localization (# of found hot nodes/pt). Both metrics define the potential for agent performance for ILM and nodal metastasis. Results: The contrast of the localization rate/pt for TcSC at 94% was based on TcSC’s reviewed value. The results for the proportion of TCP patients were localization rates of 99.91% by meta-analysis and 98.65% by pooled analysis; p<0.0001 and p<0.008, repsectively (1-sided test, H0 p=/< 0.94). The contrast of the # hot nodes/pt. This value was benchmarked at 1.6 nodes/pt based on TcSC’s review value. The results for the nodes/pt of TCP patients were 2.08 by meta-analysis and 2.16 by pooled analysis; p<0.0001 and p<0.0001, repsectively (1-sided test, H0 p=/< 1.6). Conclusions: TCP provided significantly greater performance in key metrics, node localization/pt, and degree of localization (nodes found/pt) in breast cancer. All values are highly statistically significant for the key clinical performance parameters. These data suggest that receptor targeted TCP may provide improved diagnostic ILM over the currently employed nonspecific RPA lymphatic mapping agent TcSC.
Effect of matrix metalloproteinase-2 on CD8+ T cell and NK cell responses.
Authors: Gallois Anne , Godefroy Emmanuelle , Manches Olivier , Bhardwaj Nina 
Published: March 2020

e21081 Background: Matrix metalloproteinase 2 (MMP-2) cleaves many components of the extracellular matrix and is over-expressed in several cancers including melanoma. High levels of MMP-2 expressed by tumors are associated with later tumor stages, increased dissemination and poorer survival/prognosis. We recently identified a pathway whereby MMP-2 functions as a human endogenous “conditioner” that skews CD4+ T cells towards a detrimental/inefficient TH2 phenotype through OX40L expression and inhibition of IL-12p70 production. We unraveled the underlying mechanism: MMP-2 degrades the type I IFN receptor (IFNAR1), thereby preventing STAT1 phosphorylation necessary for IL-12 production. Here, we described IFNAR1 as a novel MMP-2 substrate. This finding opens the way to characterize a possible new immunosuppressive effect of MMP-2 on other IFNAR1+ immune cells such as NK cells and CD8+ T cells. Methods: CD8+ T cells and NK cells were purified from PBMCs and incubated with MMP-2 overnight. IFNAR1 expression was assessed by FACS and Western Blot. Granzyme B, TNF alpha and IFN gamma expression was determined by intracellular staining of memory T cells. NK cells cytotoxicity was measured by Lamp-1 expression after 4h of coculture with K562 cells. Results: We first found that IFNAR1 is expressed by NK and CD8+ T cells and confirmed that MMP-2 degraded it. Memory CD8+ T cells significantly down-regulate granzyme B expression in the presence of MMP-2. Furthermore, memory CD8+ T cells pre-incubated with MMP-2 were significantly impaired with respect to IFN-gamma and TNF-alpha production. Finally, our data suggest that NK cells’ cytotoxicity might also be affected by MMP-2 through modulation of Granzyme B production. Conclusions: Our data strongly suggest that MMP-2 regulates memory CD8+ T cells not only by down regulating cytolytic activity through inhibition of granzyme B, but also by inhibiting cytokine production such as IFN-gamma and TNF-alpha. Elucidating the underlying mechanisms of MMP-2-dependent immune regulation should lead to the development of innovative immune therapies bypassing tumor escape to treat cancer patients.
Enhancement of human cancer cell migration and invasion by anaphylatoxin C5a viaaberrantly expressed C5a-receptor.
Authors: Nitta Hidetoshi , Takamori Hiroshi , Imamura Takahisa , Baba Hideo 
Published: March 2020

e21084 Background: Accumulated evidence has revealed implications of chemokines and their receptors in the pathophysiology of cancer. Anaphylatoxin C5a, a fragment of the complement 5th component (C5), is a potent chemokine for leukocytes that express the C5a receptor (C5aR). Recent reports indicate production of C5a by proteases of various origins, besides a by-product of activation of the complement system, and expression of C5aR in non-myeloid cells, suggesting an involvement of C5a in these cell biology. However, little is known about roles of C5aR in human cancer. Aim of our study was to investigate the expression and the role of C5aR on cancer cells in proliferation and invasion. Methods: Surgically resected cancer tissues and cancer cell lines were investigated C5aR expression. The effect of C5a on cancer cell proliferation, cell shape and invasion activity were examined in vitro and in vivo. Results: By immunohistochemical analysis, C5aR was strongly expressed in human cholangiocarcinoma samples. Using RT-PCR, immunoblotting and flow cytometric analysis human cholangiocarcinoma cell line MEC but not HuCCT1 were found to express C5aR. Incubation of both cancer cells with C5a did not increase these cells, indicating no mitogenic effect of C5a on these cells. Interestingly, C5a stimulated invasion of MEC and C5aR-transgenic HuCCT1 when examined by Matrigel invasion assay, which was suppressed by anti C5aR antibody. F-actin staining and human MMPs array revealed that C5a induced actin polymerization and MMPs production, respectively, in MEC and C5aR-transgenic HuCCT1. In contrast, these C5a effects were not seen in HuCCT1. Conclusions: These results illustrate a novel activity of C5a-C5aR axis that promotes cancer cell invasion through cytoskeletal rearrangement and MMP release. Thus, targeting this signaling pathway may provide a useful therapeutic option for cancer treatment.
Identification of new liver tumor biomarkers using proteomics.
Authors: Maimonis Peter J. , Zen Yoh , Britton David J. , Brand Andrew , Ward Malcolm , Pike Ian , Quaglia Alberto 
Published: March 2020

e21091 Background: It is crucial to identify new biomarkers that help diagnose cancers at an early stage and contribute to the development of new anti-cancer drugs. Here, we identify new liver tumor biomarkers using proteomics. Methods: We evaluated 9 types of liver tissue from 55 patients: normal liver parenchyma (n=7), hepatocellular carcinoma (HCC) (n=7), normal bile duct (n=6), peripheral cholangiocarcinoma (CC) (n=7), hilar CC (n=7), CC associated with primary sclerosing cholangitis (PSC) (n=7), metastatic colorectal cancer (n=7), and mixed HCC/CC after trans-arterial chemoembolization (n=7; areas of HCC and areas of CC separately examined). Protein extracted from microdissected, formalin-fixed paraffin-embedded tissue (0.15 mm3 in total) was reduced, alkylated and digested with trypsin in a stacking gel. Peptides were analyzed using nano-liquid chromatography-Mass spectrometry. The raw data were searched using Mascot. Normalized spectral counts for each protein among each tissue type were compared. For each comparison, an unrelated t-test was computed to obtain the p value. q values (adjusted p values) were computed using a direct False Discovery Rate approach (q < 0.05 considered statistically significant). Results: The mean number of proteins identified per sample was 762 +/- 119 S.D, resulting in a total of 2643 proteins identified. Numbers of proteins with significantly different expression levels among different tissue types are shown below. Some of these proteins are being reported here for the first time in the context of liver carcinogenesis. Conclusions: This study revealed proteins that are significantly over/underexpressed in particular types of liver tumors. Clinically useful new biomarkers may be present among these proteins and are now undergoing validation using immunohistochemisty. [Table: see text]
18F-fluorothymidine (FLT) PET-CT for early response assessment in advanced pancreatic cancer.
Authors: Challapalli Amarnath , Wasan Harpreet S , Al-Nahhas Adil , Aboagye Eric , Coombes Charles , Sharma Rohini 
Published: March 2020

e21093 Background: Advanced pancreatic cancer has a poor prognosis with a median survival of 6-10 months. There is a need for early non-invasive assessment of treatment response. We evaluated FLT PET-CT combined with a kinetic spatial filtering method (FLT-PETKSF) for detecting response to gemcitabine-based chemotherapy in advanced pancreatic cancer. Methods: Dynamic FLT PET-CT data were collected from patients with confirmed locally advanced or metastatic pancreatic cancer before and 2 weeks after the first cycle of chemotherapy. Changes in tumor FLT-PET variables with treatment were determined. Standardized uptake value (SUV) reduction of 18% was taken as cut-off for response. Voxel quantification of each tumor volume was performed on the filtered data. Each voxel-intensity was normalised by injected dose, body weight and decay corrected to obtain the SUV for the voxel. Changes in high intensity voxels (HiVox: SUV ≥ 2) - were computed. Results: Results of the first 5 patients are discussed. There were 4 primary and 9 metastatic tumors. FLT-PETKSF improved tumor-to-background ratio and enabled visualisation of all the primary and metastatic tumors. The mean (± SD) average and maximum SUV at 60 min (SUV60, av & SUV60, max) of the primary lesions was 2.10 (±0.38) & 4.85 (±1.55) and that of the metastatic lesions was 3.74 (±1.49) & 6.90 (±2.05) respectively. The mean (± SD) percentage reduction in the SUV60, av & SUV60, max, HIVox was 26 (±44), 18 (±38) and 23 (±50) respectively. The changes in the voxel occurrences correlated strongly with the changes in both SUV60, av & SUV60, max (Pearson r-0.9, p=0.001) .Overall, there were 2 partial responders and 3 with stable disease. These responses concurred with response evaluation on mid-treatment CT scan. Conclusions: FLT-PET and FLT-PETKSF enables visualisation of the pancreatic tumors and the liver metastases, and could be used to monitor response to therapy.
Clinical significances of ErbB family receptors, Myc, and mitogen-activated protein kinase (MAPK) in patients with early-stage non-small cell lung cancer.
Authors: Ludovini Vienna , Bellezza Guido , Bianconi Fortunato , Ferri Ivana , Pistola Lorenza , Flacco Antonella , Baldelli Elisa , Chiari Rita , Minotti Vincenzo , Tofanetti Francesca Romana , Siggillino Annamaria , Ragusa Mark , Vannucci Jacopo , Puma Francesco , Sidoni Angelo , Crinò Lucio 
Published: March 2020

e21094 Background: EGFR deregulation has been extensively studied in non small-cell lung cancer (NSCLC), but less is known about the expression and role of other ErbB receptors and their downstream signal transductions. Myc and MAPK are key downstream components of the EGFR pathway and have significant roles in cell survival, proliferation, and growth. The purpose of this study is to determine the prognostic value of EGFR, ErbB2, ErbB3, ErbB4, Myc and MAPK by immunohistochemistry (IHC) in early stage NSCLC. Methods: 109 NSCLC were evaluated: median age was 67 years (range 40–84); Male/Female:93/16; squamous (SCC)/adenocarcinoma (ADC)/BAC/other: 52/36/3/18; smoker/never smoker:100/9, and stage I/II/III:67/17/25. The tumors with ≥10% positive cells were classified positive, further confirmed by Receiver Operating Characteristic (ROC) analysis. Kaplan-Meier estimates of survival and time to recurrence were calculated for clinical and biologic variables using Cox model for multivariate analysis Results: EGFR and EbrB3 were associated with SCC (p<0.0001 and p=0.004, respectively) whereas ErbB2 and Myc with ADC (p=0.004 and p<0.0001, respectively) . EGFR and ErbB3 were significantly associated (p=0.003), as well as MAPK and ErbB4 (p=0.02). At a median follow-up of 75 months the contemporary overexpression of EGFR, ErbB2 and MAPK was associated with shorter disease free survival (DFS) (HR=5.4, p=0.002) and overall survival (OS) (HR=8.9, p<0.0001). At multivariate analysis adjusting for stage, the co-expression of EGFR, ErbB2 and MAPK was an independent predictor for worse DFS and OS (HR=5.7, p=0.004; HR=8.67, p<0.001, respectively). Conclusions: Our results suggest that in early stage NSCLC, the co-expression of EGFR, ErbB2 and MAPK predicted a worse prognosis. ErbB3, ErbB4 and Myc were not prognostic factors. Such features may have important implications for future targeted therapies. We thank Italian Association for Cancer Research (AIRC) for supporting the study.
The value of RCAS1 as a potential biomarker in non-small cell lung cancers.
Authors: Tsoukalas Nikolaos , Kostakis Ioannis , Siakavellas Spiros , Tolia Maria , Papakostidi Aristoula , Karameris Andreas , Tzovaras Alexandros , Sfiniadakis Ioannis , Tsiambas Evangelos , Ardavanis Alexandros , Manolis Evangelos , Kittas Christos , Theocharis Stamatios 
Published: March 2020

e21098 Background: RCAS1 (Receptor-binding Cancer Antigen expressed on SiSo cells) is a membrane protein that is expressed in different types of cancer. It halts the cell cycle and/or induces the apoptosis of the immune system cells within the tumour microenvironment. Hence, it is possible that this molecule is involved in the mechanism of the tumour cells’ escape from the immune system surveillance (immunoescape). Methods: Patients with primary non small cell lung cancer, initially eligible for surgical treatment, were included. The tissue samples (paraffin cubes) were processed using the Tissue Micro-arrays Method. Immunohistochemical study was done with specific monoclonal antibodies for RCAS1 and Ki-67. The image analysis was feasible due to a special program (image analysis software). In addition, a database was created that included the clinical and pathological characteristics of the patients. After the surgery patients received chemotherapy and/or radiotherapy according to the guidelines. Associations between variables were analyzed with non-parametric tests (Mann-Whitney and Kruskal-Wallis test) and the survival analysis with the Cox proportional hazards model. Two tailed p values ≤ 0.05 were considered to be statistically significant. Results: In total, 112 patients were examined (93 men and 19 women), mean age 63,6 years old. Almost 53% of the cases were adenocarcinoma, 33% squamous cell, 9% large cell, and 5% other types. Statistical significance was identified correlating RCAS1 over-expression to lower overall survival (p=0.032), to grade III (p=0.006) and in a positive correlation between RCAS1 and Ki-67 (p<0.001). Moreover, there is a trend of RCAS1 over-expression in advanced or metastatic stages and in tumours with lymphovascular invasion. In contrast, protein expression was not strongly associated to tumour size, to histological type, to patient age or to gender. Conclusions: The most important conclusions of this study are the negative correlation between RCAS1 and overall survival, the over-expression of RCAS1 in grade III tumors and the positive correlation between RCAS1 and Ki-67 which means that when Ki-67 increases RCAS1 is higher. Consequently, this molecule RCAS1 probably could be considered as a marker of the tumor’s aggressiveness.