bluuebunny/crossref_metadata_2025_split huggingface dataset:
Metabolic intervention targeting 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) using a structure-based design.
Authors: Ährlund-Richter Lars , Färnegårdh Katarina , Wiita Elisee , Jönsson Mattias , Norström Carina , Martinsson Jessica , Hallberg Kenth , Ringom Rune , Mellstedt Hakan 
Published: March 2020

e13518 Background: By producing fructose-2,6-bisphosphate, PFKFB3 functions as an activator of anaerobic glycolysis. PFKFB3 is both over expressed and over activated in many of the types of human cancer. Specific inhibition of the PFKFB3 enzyme results in a reduction in metabolism and cell growth in oxygen-deficient cancer environments. Methods: High-throughput screening. Medicinal Chemistry. Structure-Based Drug Design, X-ray Crystallography. NMR. Isothermal Calorimetry. Dynamic Light Scatttering. ADME. Results: A high-throughput screening of 50.000 selected compounds, by means of a biochemical assay, generated 105 hits including both ATP-and non-ATP competitive hits as identified by NMR binding experiments. The latter type was prioritized and two hits with a similar “ring-linker-ring structure” were selected for further expansions. Interestingly, although structurally similar, the two hits were found by means of X-ray crystallography to exhibit different binding modes within the fructose pocket. Based on their respective binding mode, two chemical series were developed displaying different ADME properties and PFKFB isoenzyme selectivity. Calorimetry verified a reversible strong enthalpy driven, direct binding for both chemical series. A third chemical series was developed towards yet another unoccupied binding pocket within the fructose-site, yielding a 5-fold increase in potency. Strong interactions within the new pocket were confirmed using X-ray crystallography. Our PFKFB3 inhibitors were shown to reduce tumor cell growth in vitro and to exhibit combinatory effects with Cisplatin. Conclusions: We have targeted the fructose-binding pocket of PFKFB3, developed compounds with nM binding potency and have gained a detailed understanding of SAR via structural information. The structure-based analysis has provided a good understanding of the molecular interactions, which is important for further biological/clinical positioning: e.g., combination with chemotherapy, optimization of PK properties and proof of principle in vivo.
FDG-PET based dose escalated adjuvant tomotherapy (image-guided intensity-modulated radiotherapy) utilizing 4D-CT planning for high-risk adrenocortical carcinoma (ACC): A case study.
Authors: Mallick Ujjal , Ramamurty S , Carr Ruth Elizabeth , Willis Nick , Ball S , Bliss R , Mott Judith H. , Wilsdon John B. , Johnson S 
Published: March 2020

e13522 Background: ACC is a rare endocrine malignancy with an annual incidence of 0.5-2/million/year. Surgery is the mainstay of treatment but relapse rate is high and the prognosis is dismal (20-50% 5 year survival). Adjuvant Radiotherapy improves local control in cases with high risk of local failure and dose escalation may achieve better results. Dose Escalation with 3-Dimensional computed tomography (CT) based radiotherapy is difficult for ACC due to unacceptable toxicity to several adjacent organs at risk (OARs) e.g., the kidneys, stomach bowel, liver, spinal cord. FDG-PET helps delineation of areas of higher metabolic activity and density of clonogenic cells within the target (Biological Target Volume or BTV) where focal dose escalation could be directed safely for improving local control as in other solid tumours. Tomotherapy (IMRT with daily image guidance) delivers precision radiotherapy with integrated boost to the BTV, lower dose to OARs, improved coverage of the targets, and better dose delivery across the targets which are essential for tumour control. 4-Dimensional CT for planning allows adequate Clinical Target Volume(CTV) coverage despite respiratory movements. Methods: A 63 year old lady had left adrenalectomy for an adrenocortical carcinoma measuring 29 x 18 x 25mm (TNM pT1NXMX) thought to be microscopically incompletely excised (R1) with a proliferation index ( Ki-67) of 40% for which adjuvant radiotherapy was advised. PET-CT, 4D-CT and Tomotherapy were used for planning. A total dose of 50.4Gy to PTV with a simultaneous integrated boost to BTV taking it up to 59.40Gy (1.98Gy/ fraction) in 30 daily fractions was prescribed. Results: Target dose and coverage were very satisfactory. The doses to OARs were well within tolerance despite dose escalation; she completed treatment with only grade- fatigue. She had metastatic disease at 11 months follow-up without local relapse. Conclusions: To our knowledge this is the first report of using FDG –PET based dose escalation, 4D CT and Tomotherapy in high risk ACC to improve local control and needs further study.
Inhibition of tumor cell growth, invasion, and metastasis by SIM-89, a novel inhibitor of HGF receptor tyrosine kinases.
Authors: Pei Jun , Han Baohui , Shao Minhua , Sha Huifang 
Published: March 2020

e13528 Background: The Met receptor tyrosine kinase and its ligand, hepatocyte growth factor (HGF), are overexpressed and/or activated in a wide variety of human malignancies. SIM-89, developed by Simcere Bio-Pharmaceutical Co, Ltd, is a small-molecule kinase inhibitor that targets members of the HGF receptor tyrosine kinase families. Methods: Kinase inhibition was investigated using Z-lyte detection technique(Invitrogen), LanthaScreen Tb-activity technique and LanthaScreen Binding technique. Inhibition of p-Met and Met is validated through Westernblot. IC50 of the compound is determined by CCK-8. HGF level in culture medium is determined by ELISA.Migration and invasion assayare done with Transwell system. A549 and H460(2×106)are implanted subcutaneous in right upper extremity of nude mice.Once daily oral administration of SIM-89/ placebo are given to nude mice for 35d.Tumor growth curve is calculated. B16F10 tumor cells (2 × 105) were implanted via i.v. tail vein injection into mice on day 0. SIM-89/ placebo administration was initiated 3 days after implantation for 10 days followed by assessment of lung tumor burden. Lung nodule diameters were morphometrically measured on digitally captured images. The results for each treatment group (n = 10 animals) were averaged, and statistical t test analysis was done comparing each treatment group to the placebo treated control. Results: SIM-89 is a small-molecule kinase inhibitor that targets members of the HGF receptor tyrosine kinase families,with additional inhibitory activity toward AMPK (A1/B1/G1) and TRKA. SIM-89 significantly decreases HGF level in culture medium of nsclc cell lines at the IC50 of 625nmol/l. In vivo, these effects produce significant dose-dependent inhibition of tumor burden in an experimental model of lung metastasis. Once daily oral gavage administration of SIM-89 resulted in a dose-dependent reduction in tumor burden, as determined by a reduction in lung wet weights. Conclusions: These data indicate that SIM-89 may prevent tumor growth through a direct effect on tumor cell proliferation and by inhibition of invasion mediated by HGF receptors.
Single nucleotide polymorphisms analysis of BRAC and ERCC1 as predictor of recurrence after chemoradiation for cervical cancer patients.
Authors: Féliz Luis Roberto , Pereira Verónica , Monzo Mariano , Munoz Carmen , Fuste Pere , Gascon Pedro , Torrents A. , Vidal Boixader Laura 
Published: March 2020

e13540 Background: A number of patients with locally advanced cervical cancer will recur despite presenting initial response to chemoradiation. Single-nucleotide polymorphisms (SNP) of DNA repair genes have been found to be predictors of efficacy of chemotherapy and radiotherapy. Methods: We evaluated the presence of SNP in ERCC1, BRAC 1 and 2 genes. We analysed paraffin-embedded biopsies from patients who had relapsed after receiving treatment with chemoradiation, for SNP of the mentioned genes. The status of the alleles wild type (wt) or at least 1 SNP was compared with time to progression (TTP) and toxicity. Results: 90 patients who experience recurrence of their cervical cancer were included in the analysis. Of those, we only could obtain evaluable tumour from 43 patients. Median age: 52.5 yrs (31-81). Histology: 32 squamous cell, 8 adenocarcinoma, 3 adenosquamous. One SNP in BRAC1 (rs12516) was found to be significantly associated with better TTP for the mutant variant compared to the wild-type (124 m. vs. 14 m.). Furthermore, all six patients who presented with severe (grade 3-4) toxicity had this wild-type SNP (rs12516) in BRAC1 gene. Conclusions: We have identified a SNP which confers better outcome in patients with cervical cancer. Further analysis need to be done to determine its relation to radiation-induced toxicity in this group of patients.
Correlation of lactate dehydrogenase (LDH) isoenzyme profile with outcome in advanced colorectal cancer (CRC) patients (pts) treated with chemotherapy and bevacizumab (BEV) or cediranib (CED).
Authors: Bar Jair , Spencer Stuart , Morgan Shethah , Pike Laura , Cunningham David , Robertson Jane D , Jürgensmeier Juliane M , Goss Glenwood D. 
Published: March 2020

e13541 Background: There is a clinical need for predictive biomarkers of efficacy of VEGF signaling inhibitors (VSIs). LDH is a tetramer that can include any combination of the M and H subunits. LDH4 and 5 isoenzymes are composed mostly of the M subunit and are more abundant in hypoxic conditions. We speculated that the levels of LDH isoenzymes in pts serum may correlate with outcome of pts treated with VSIs. HORIZON I trial enrolled pts that were randomized to mFOLFOX6 with BEV or CED. A retrospective exploratory analysis was performed on baseline serum samples of these pts. Methods: Total serum LDH was tested on fresh samples during the conduct of the trial. About 2 years after the trial, frozen samples stored at -70 degrees were tested for total serum LDH and LDH isoenzymes, measured by agarose electrophoresis and a colorimetric enzymatic assay. Relative levels of M and H subunits were derived based on the known structure of each isoenzyme. Progression free survival (PFS) and overall survival (OS) were compared by subgroups of total LDH and LDH isoenzyme levels. P values were not calculated for these exploratory analyses. Results: Baseline total LDH levels from fresh serum were available for 207 pts. Total and isoenzyme LDH levels were available for frozen serum samples of 189 pts. Total LDH in the frozen and fresh samples correlated (R=0.9). Distant isoenzymes (e.g. LDH1 and 5) were negatively correlated. High M/H subunits ratio correlated with poor OS (HR=1.804, 95%CI 1.24-2.620). A non-significant trend for better OS to CED-treated vs. BEV-treated pts was seen in pts with high M/H ratio (e.g., CED 30mg vs BEV HR=0.685, 95%CI 0.382-1.23). Conclusions: Evaluation of LDH isoenzymes is feasible using serum samples kept frozen for 2 years. A negative correlation is seen between hypoxic-related and oxic-related isoenzymes. In CRC pts treated with a VSI, LDH isoenzyme hypoxia-associated profile correlates with poorer outcome. LDH isoenzyme profile as a possible predictive biomarker for benefit from CED vs. BEV requires further investigation.
The effect of BMI-1 inhibition on brain cancer stem cells.
Authors: Mehta Monal , Khan Atif J. , Sabaawy Hatem E. , Haffty Bruce George 
Published: March 2020

e13543 Background: Glioblastoma (GBM) is the most frequent and deadly brain cancer. Despite tolerance doses of radiation, control of tumor growth within the brain remains a formidable failure. Since the identification of brain cancer stem cells (BCSCs), efforts are underway to target the pathways regulating these cells. The role of Bmi-1 (B-cell specific MMLV insertion site-1), a polycomb member of chromatin-remodeling complex, in BCSCs self-renewal was elucidated. Here we utilize shRNA targeting or pharmacological inhibition of Bmi-1 in GBM cell lines and primary cells as a radiosensitizer to examine the effects of combination therapy on cell death and BCSCs differentiation. Methods: Cells were pre-treated with a Bmi-1 inhibitor before being irradiated. Serial neurosphere assay, a measure of self-renewal potential, was employed to study the effects of radiation, Bmi-1 inhibition, or the combination on BCSCs. The efficacy of this combination on cell death was assessed with MTT and clonogenic assays. Next, the abilities of the inhibitor and radiation to induce differentiation in GBM cell lines and primary cells were quantified. Further, by utilizing a novel zebrafish orthotropic xenograft model, small molecules targeting Bmi-1 and other BCSC pathways can be identified, and used to predict response to combination therapies. Results: Targeting of Bmi-1 in combination with radiation, specifically as a radiosensitizer, induced significant cell death in GBM cells, and was five-fold more effective than radiation only. Importantly, the neurosphere forming ability of BCSCs was severely compromised when the cells were treated with the combination, indicating a potent effect on the stem cell constituency. These effects may be due to loss of BCSC self-renewal potential, increased differentiation, and/or apoptosis as cells treated with the combination exhibited decreased expression of neural stem cell markers and abnormal phenotypes compared to single treatment. Conclusions: Targeting of Bmi-1 may eliminate the subpopulation of radioresistant BCSCs. Bmi-1 inhibition when combined with radiotherapy might provide an effective therapy for GBM patients specifically through its effect on BCSCs by affecting their survival, proliferation, and stem cell features.
Effect of small inhibitors of nuclear export (SINE) on growth inhibition and apoptosis of human melanoma cells.
Authors: Lesinski Gregory B. , Yang Jennifer , Bill Matthew A , Landesman Yosef , Shacham Sharon , Kauffman Michael , Senapedis William , Kashyap Trinayan , Saint-Martin Jean-Richard , Kendra Kari Lynn 
Published: March 2020

e13549 Background: Inhibition of nuclear export can promote re-activation of tumor suppressor pathways. CRM1 (chromosomal regional maintenance 1) or XPO1 (exportin 1) is the major protein that mediates nuclear export. We hypothesized that CRM1 mediated nuclear export represents a novel therapeutic target that can be manipulated to inhibit melanoma cell survival. Methods: The growth inhibitory and pro-apoptotic effects of KPT-185, KPT-276 and KPT-330, small molecules selective inhibitor of nuclear export (SINE) were evaluated in human melanoma cell lines using an MTT assay and Annexin V/PI staining, respectively. Fluorescence microscopy and immunoblots were used to assess nuclear accumulation of tumor suppressor proteins. The trans-isomer of KPT-185 and DMSO (vehicle) were used as a negative controls in all assays. The pharmacokinetic (PK) profile of all compounds was evaluated in mice. Results: CRM1 protein was highly expressed in human melanoma cell lines with diverse molecular profiles (i.e., B-Raf, NRAS, p53). KPT-SINE inhibited melanoma cell growth in a concentration-dependent manner and induced apoptosis at nanomolar concentrations. Importantly, there was no evidence that B-Raf V600 mutational status influenced melanoma cell response to these agents. Nuclear accumulation and/or induction of p53, p21, FOXO3a, STAT1 and BAD, and reduction of MCL-1 occurred in melanoma cells at time points prior to apoptosis as shown by increase in cleaved PARP and caspase 3 levels. PK studies were conducted in mice following oral administration of 10 mg/kg, to guide drug selection for our ongoing efficacy studies in murine melanoma models. KPT-185 showed limited bioavailability and systemic exposure, while KPT-276 and KPT-330 showed >50% bioavailability reaching Cmax >5µM. Conclusions: This study represents the first report of CRM1 inhibition in melanoma. These data indicate that the novel SINE compounds can effectively inhibit CRM1-mediated nuclear export and induce apoptosis in melanoma cells. KPT-330 is currently under development as orally bioavailable, small molecule inhibitors for a human clinical trial.
The regulation of stem-like property in gastric cancer cells mediated by tumor associated macrophage.
Authors: Ishimoto Takatsugu , Watanabe Masayuki , Iwatsuki Masaaki , Ida Satoshi , Nagai Yohei , Iwagami Shiro , Baba Yoshifumi , Sakamoto Yasuo , Miyamoto Yuji , Baba Hideo 
Published: March 2020

e13553 Background: CD44 has recently been identified as one of the cell surface markers associated with cancer stem cells (CSCs) in several types of tumor. We have reported the functional role of CD44 variant isoform in the maintenance of low reactive oxygen species (ROS) levels in gastrointestinal cancer cells. Previous studies demonstrated that tumor associated macrophage promotes tumor progression, but the functional role for macrophage in the regulation of CSC marker expression is not known. Methods: We analysed the relationship of CSCs and macrophage infiltration in gastric cancer cell lines and human gastric cancer samples. Results: Here, we showed activated macrophage by LPS and IFNγ produces inflammatory cytokines and chemokines, and these macrophage express CD68 and CD163, which are known as specific markers for pan-macrophage and M2 macrophage. we found that co-culture with activated macrophage triggers the CD44 and Bmi1 expression in gastric cancer cells and promote stem-like property that possess sphere-forming ability. Furthermore, we investigated the relation between CD44 expression and infiltrated macrophage in human gastric cancer. The staining patterns of CD44, Bmi1 and macrophage specific markers (CD68 and CD163) were significantly correlated in tumorous tissues of human gastric adenocarcinoma. Conclusions: These findings establish inflammatory cytokines, which producted from activated macrophage, induce the CD44 and Bmi1 expression in gastric cancer cells. These findings revealed that a role for tumor associated macrophage in the expansion of gastric cancer stem-like cells and subsequent malignant progression.
The intraductal injection of 5-fluorouracil for breast cancer: Preventive effect and immune response.
Authors: Mori Tsuyoshi , Wada Satoshi , Olino Kelly , Edil Barish H , Pardoll Drew M. , Schulick Richard D. , Sukumar Saraswati , Yoshimura Kiyoshi 
Published: March 2020

e13559 Background: Chemotherapy has been shown to act as an immune response modifier. Many groups are using chemotherapy to induce immune responses such as by using low doses of cyclophosphamide. Building up on our previous work (Murata et al., 2006, Stearns et al., 2011), we demonstrate that intraductal (i.duc), but not the intravenous (IV) route of injection of chemotherapy to HER2/neu transgenic mouse mammary glands (MMG) alters the tumor environment to effectively induce immune effector cells. Methods: 1.Tumor-free survival: 5-fluorouracil (5FU) was administered intraductally to the MMGs on the left side of mice; the MMG on right side were untreated. 2. FACS analysis: 20w mice were administered 5FU 2 times in a 4 week interval i.duc only to the left side, IV, or no treatment (NT). At week 5, the mice were sacrificed and the regional lymph nodes (RLN) and spleen were removed. Lymphocytes from RLN and spleen were analyzed. 3. Tumor re-challenge: GT8 cancer cells were injected into the one side of 4th mammary gland fat pad of 12 week mice. The mice were then injected i.duc with 5-FU to all teats in the tumor implanted side, or IV or given NT. The surviving mice were re-challenged with GT8 cancer cells into the ipsilateral 3rd mammary gland fat pad, 4 weeks after the first inoculation. Results: 1. The tumor incidence in the 5FU treated side after 16 weeks was significantly lower (13% of 70 glands) compared to mice that received NT (34% of 150, p=0.001), and IV (28% of 70, p=0.03). The tumor incidence in the untreated side of 5FU-treated mice was also significantly lower (18% of 70) compared to the NT (p=0.01). 2. The number of CD8 T cell showed no change among the groups in the RLN but was significantly lower (p=0.01) in the spleen of the i.duc. The number of CD62LLOW+ T cell in treated side of RLN was significantly higher compared to the IV (p=0.02) and NT (p=0.02). The number of CD62LLOW+ was low in the spleen. 3. The survivors in the group previously treated with i.duc 5-FU were able to achieve a lasting rejection of the tumor re-challenge when compared to mice who previously received 5-FU IV. Conclusions: I.duc administration of 5FU into MMG effectively induced immune effector cells and prevented mammary tumor growth in HER2/neu transgenic mice.
The effect of the CXCR7 inhibitor CCX662 on survival in the ENU rat model of glioblastoma.
Authors: Ebsworth Karen , Walters Matthew J. , Ertl Linda S , Wang Yu , Zhang Penglie , Powers Jay P , Sullivan Timothy J. , Jaen Juan C. , Brown Martin , Schall Thomas J. 
Published: March 2020

e13580 Background: Glioblastoma (GBM) is the most common form of malignant brain cancer. Despite aggressive therapy, consisting of radiotherapy, surgical resection, and chemotherapeutic treatment, mean survival from time of diagnosis remains little more than one year. The chemokine receptor CXCR7 is highly expressed on both human glioma cells and tumor-associated vasculature, and may play a key role in tumor growth and survival. Methods: The in vitro activity of the CXCR7 antagonist CCX662 was determined using radioligand binding assays and trans-endothelial migration assays. The therapeutic effects of CCX662 were investigated in the ENU-induced rat model of GBM in combination with radiotherapy. Results: CCX662 is a highly potent and selective small molecule inhibitor of CXCR7. CCX662 inhibits the binding of 125I-CXCL12 to CXCR7 with an IC50 of 9 nM in buffer, and displays minimal serum shift with an IC50 of 18 nM in 100% human serum. CCX662 inhibits the CXCR4-directed trans-endothelial migration of CXCR4+/CXCR7+ NC37 cells towards CXCL12 (SDF1) with an IC50 of 106 nM. CCX662 also potently inhibits binding of 125I-CXCL12 to rat CXCR7 in the presence of 100% rat serum with an IC50 of 14 nM. In vivo inhibition of CXCR7 with CCX662, in concert with radiotherapy, results in a significant extension of survival time in the ENU-induced rat model of GBM. The median survival time for rats (reflective of GBM tumor progression) treated with the combination of irradiation and CCX662 was 234 days compared with 160 days for untreated rats (p<0.01) or compared with 174 days for rats given irradiation plus vehicle (p<0.01). As expected for a non-cytotoxic drug, dose-range finding 14-day toxicology studies in rats and dogs have demonstrated large (> 10 and 25-fold, respectively) safety margin relative to highly effective levels of the drug. Conclusions: CCX662 is a highly selective and potent small molecule inhibitor of CXCR7 with profound, therapeutic benefit in an aggressive rodent model of GBM. These data indicate that inhibition of CXCR7, using CCX662, may be a promising strategy for the treatment of glioblastoma.