bluuebunny/crossref_metadata_2025 huggingface dataset:
Catumaxomab with and without prednisolone in patients with malignant ascites due to epithelial cancer: Results from the phase IIIb CASIMAS study.
Authors: Sehouli Jalid , Wimberger Pauline , Vergote Ignace B. , Rosenberg Per , Schneeweiss Andreas , Bokemeyer Carsten , Salat Christoph , Scambia Giovanni , Berton-Rigaud Dominique , Siena Salvatore , Santoro Armando , Cervantes-Ruiperez Andres , Guastalla Jean Paul , Tournigand Christophe , Colombo Nicoletta , Dudnitchenko Alexander Serg , Bondar Vladimir , Westermann A.M. , Lordick Florian
Published: March 2020

e13097^

Background: The primary objective of this study was to compare catumaxomab with prednisolone (CP) to catumaxomab without prednisolone (C) as 3-hour intraperitoneal (i.p.) infusion by demonstrating superiority for safety and non-inferiority for efficacy of the CP arm. Methods: 219 patients were randomized to catumaxomab plus premedication of 25 mg prednisolone (111 pts) or to catumaxomab alone (108 pts). The primary endpoint was the composite safety score (CSS) summarizing the worst CTCAE grades for the main TEAEs (pyrexia, nausea, vomiting, and abdominal pain). A potential impact of prednisolone on efficacy was assessed by the co-primary endpoint puncture-free survival (PuFS). Further parameters included overall survival (OS) and time to next therapeutic puncture (TTPu). Results: The superiority of CP for safety was not proven as the mean CSS was comparable in the two groups (CP: 4.1; C: 3.8 for; p= 0.383). The median PuFS was slightly lower in CP (30 days) compared to C (37 days). However the hazard ratio (HR) for PuFS (HR: 1.130, p=0.402) as well as the 75% quartiles (CP: 155 days, C: 92 days) were in favour of CP compared to C. The median TTPu was similar in both groups (CP: 78 days; C: 102 days, p= 0.599). The majority of patients (123 pts) had no therapeutic paracentesis prior to death (CP: 54.8%; C; 61.7%, p=0.297). Median OS was longer for CP (CP: 124 days; C: 86 days, p= 0.186) without statistical significance. Conclusions: The CASIMAS results are in concordance with the data of the pivotal study and thus confirm the robustness of the treatment effect of catumaxomab in malignant ascites. The administration of 25mg prednisolone as premedication prior to catumaxomab infusion did not change the safety profile and did not negatively impact the efficacy of catumaxomab. The composite safety score after 3-hour infusion time was comparable to that seen in the pivotal study using 6 hours.
Results of the first interim analysis of the CARMA study to investigate efficacy and safety profile of catumaxomab in clinical practice.
Authors: Kunzmann Volker , Sehouli Jalid , Schmalfeldt Barbara , Wimberger Pauline , Kurbacher Christian M. , Tempelhoff G.-F. , Vanhoefer Udo J. , Ebeling P. , Breuer F. , Schulz Holger , Welslau Manfred , Finas Dominique , Sagasser Jaqueline , Kiehl M. , Fruehauf S.
Published: March 2020

e13100^

Background: The trifunctional antibody catumaxomab is approved in the EU for intraperitoneal (i.p.) treatment of malignant ascites in patients (pts) with EpCAM-positive carcinomas. Clinical data for catumaxomab are based on the pivotal trial and several phase I/II trials. So far, the routine use of catumaxomab in clinical practice has not been evaluated systematically. Therefore, a large prospective observational study was started in 2010. The study investigates the administration of catumaxomab in a total of 160 pts with malignant ascites under routine conditions in daily clinical practice. Here we show the results of the first pre-planned interim analysis. Methods: The analysis included 49 pts with malignant ascites due to EpCAM-positive carcinomas treated with i.p. catumaxomab under routine conditions in clinical practice. Participating centres were hospitals and practices of oncologists in Germany. Primary endpoint was puncture-free interval (PFI), secondary endpoints included safety and overall survival (OS). Results: At inclusion into the study pts already had undergone a median number of 9 ascites punctures. Malignant ascites had been diagnosed for the first time 3.3 months before (median). Patients suffered from typical ascites symptoms as follows: swelling (79.6%), pain (44.9%), dyspnea (26.5%), anorexia (26.5%), obstipation (12.2%). 49 pts were treated with catumaxomab, of whom 30 pts received all 4 infusions (61.2%). Median PFI was 108 days (d), the median OS was 102 d. Most frequent adverse events were fever, nausea and diarrhoe. Conclusions: This is the first systematic report on routine use of catumaxomab in clinical practice.. Although The study population mainly comprised patients with advanced tumor diseaset a clinical benefit after catumaxomab therapy could be demonstrated. The treatment showed an acceptable safety profile Theseresults are consistent with the data of the pivotal trial. The data will be evaluated in further analyses including larger patient numbers.
A phase Ib study of CUDC-101, a multitargeted inhibitor of EGFR, HER2, and HDAC, in patients with advanced head and neck, gastric, breast, liver, and non-small cell lung cancer.
Authors: Fu Siqing , Nemunaitis John J. , Bessudo Alberto , Bauman Julie E. , Hamid Omid , Witta Samir E. , Dy Grace K. , Lai Cheng , Laliberte Robert , Voi Maurizio
Published: March 2020

e13101

Background: CUDC-101 is a small molecule EGFR/HER2 receptor tyrosine kinase and HDAC inhibitor that has shown promising activity in cancer cell lines and tumor xenograft models. A previous Phase 1 study showed 275 mg/m2 to be the maximum tolerated dose when given daily for 5 days every 2 weeks (QDx5). Dose limiting toxicity included transient creatinine elevation. This expansion Phase 1b study was conducted to further evaluate safety and tolerability of the QDx5 and of a three dose per week (3x/week) schedule for 3 weeks every 4 weeks. Methods: Eligible patients (pts) ≥18 years with advanced breast, gastric, head and neck, NSCLC or liver cancer (n = 10/tumor type); adequate marrow and organ function; and an ECOG PS ≤2 were assigned to receive CUDC-101 (275 mg/m2) on QDx5 or 3x/week schedule. NSCLC patients required documented EGFR exon 19 or 21 mutation/deletion with recent acquired resistance to erlotinib. Study parameters included safety (CTCAE v.3), antitumor response (RECIST v1.1), PK (blood and urine) and PD (tumor biopsy and CTC) assessments. Results: A total of 46 pts were treated (23/dose schedule), including 25 (54%) males, mean age 62 yrs, PS 0/1/2 (33/56/11%), and a median of 3 prior regimens (range, 0-7). The overall incidence and severity of treatment-related AEs were similar between the 2 dose schedules, including fatigue, transiently increased creatinine, nausea, vomiting, decreased appetite, anemia and asthenia, each occurring in <20% of all pts and mostly Grade ≤ 2. Grade 3 AEs were reported in 4 pts. Overall, 15 (33%) pts had SD as their best response: 4 pts had SD for >14 weeks, including 1 with head and neck (QDx5), and 2 with liver (1 lasting 47 weeks) and 1 gastric cancer (all 3 on 3x/week). Paired tumor biopsies were available for one pt showing target inhibition. CTC and PK analyses are ongoing. Conclusions: CUDC-101 was well tolerated using either dosing schedule, with preliminary evidence of antitumor activity. The more convenient 3x/week schedule was selected for an ongoing Phase 1 combination study.
Designing a web application for personalized medicine trials.
Authors: Watt Stuart , Jiao Wei , Brown Andrew M. K. , Petrocelli Teresa , Tran Ben , Zhang Tong , Dancey Janet , Siu Lillian L. , Stein Lincoln D. , Ferretti Vincent
Published: March 2020

e13107

Background: Clinical genomics uses information from a patient's genome in clinical decision-making, an example of personalized medicine. The OICR/UHN Genomics Cohort Study is assessing the feasibility and developing standard operating procedures for clinical genomics in late stage cancer patients to enable larger trials. Tumor DNA from consenting patients is sequenced across 19 genes to identify actionable mutations and inform use of targeted therapeutic agents. Informatics systems are critical as the study involves 40 staff in 5 cancer centres, 2 laboratories, 3 genomics technologies, and spans screening, consent, obtaining/processing biopsy samples, genomic analysis, clinical laboratory verification, and reporting for decision-making. Methods: We used a process-centered method to develop a web system to manage study activities. Initially it tracked patients, samples, genomic results, decisions and reports across the cohort, monitored progress and sent reminders, working alongside an electronic data capture (EDC) system for the trial's clinical and genomic results. We later added a system to read, store, and analyze the genomics data, and a knowledge base of mutations’ tumor frequency (from the COSMIC database) annotated with clinical significance and drug sensitivity to generate reports for clinicians. Results: The web tracker proved highly adaptive. The design method allowed procedural refinements mid-study, including changes in sample preparation, sample sources, and differences in nomenclature across technologies. As the study procedures stabilized, the system provided deeper support for clinical decision making, enabling the generation of draft reports for verification by an expert panel prior to forwarding to the treating physician. The web tracker complemented the EDC system with its fixed modules for collection of clinical data and genomic results. Conclusions: The system effectively complemented clinical trial software. An agile development process enabled procedures to be refined as feasibility issues were found and resolved, and enabled flexible analysis of mutation data. Our design approach helped stabilize effective procedures for a clinical genomics service, and establish means to assess its performance.
Catumaxomab: Pharmacodynamic and pharmacokinetic data from phase IIIb CASIMAS study in patients with malignant ascites.
Authors: Rosenberg Per , Dudnitchenko Alexander Serg , Bondar Vladimir , Salat Christoph , Havsteen Hanne , Bokemeyer Carsten , Berton-Rigaud Dominique , Durando Xavier , Vergote Ignace B. , Wimberger Pauline , Lordick Florian , Lindhofer Horst , Martinius Holger , Sehouli Jalid
Published: March 2020

e13108^

Background: Catumaxomab (anti-EpCAM x anti-CD3) is approved in the EU for the i.p. locoregional treatment of malignant ascites (MA). The peritoneal cavity invaded by epithelial tumor cells provides an environment for tumor-specific, EpCAM-targeted catumaxomab immunotherapy. In order to monitor the local and systemic pharmacodynamic/ pharmacokinetic (PD/PK) profile of catumaxomab, plasma and ascites samples from patients in the CASIMAS study were investigated. Methods: Plasma and ascites samples were collected in patients prior, during and after catumaxomab i.p. infusions. Samples were analysed for a) Selected cytokines (IL-2, -4, -6, -8, -10, and -12, IFN-gamma, TNF-alpha, and GM-CSF) in 20 patients, b) catumaxomab drug levels in 31 patients (14 patients with MA who received all 4 infusions of 10, 20, 50, and 150 µg were evaluable for systemic PK analysis), c) number of EpCAM+ tumor cells in ascites in 11 patients. Results: Drug levels were highest in ascites fluid ranging from 0 to 219.4 ng/mL at day 10 (n=31/31 patients). Low systemic plasma levels were detected in 7/14 patients (median cmax: 623 pg/mL). In 7/14 subjects no catumaxomab was detected systemically. Furthermore, cytokine concentrations in ascites were higher compared to plasma. Mainly IL-6, -8 & -10 and IFN-gamma were detected; in few cases TNF-alpha and IL-2. Whereas IL-4, -12, and GM-CSF were not detected at all. Anti-tumor activity was proven by i.p. tumor cell elimination. Tumor cell count in ascites dropped markedly below baseline. In the majority (7/11) of patients no EpCAM+ tumor cells were detectable after therapy. Before therapy, in 165/166 evaluable patient samples EpCAM+ tumor cells were detected in the ascites fluid. Conclusions: Catumaxomab represents a locoregional immunological i.p. treatment eliminating tumor cells in the peritoneal cavity and thereby effectively controlling MA. Catumaxomab primarily remains in the peritoneal cavity with low systemic exposure. In line with this, cytokines are mainly released locally, at the site of action. These PD/PK results confirm the results obtained in the pivotal phase II/III study.
Folate-targeted iron oxide nanoparticles loaded with cisplatin for imaging and therapy in head and neck squamous cell carcinoma (HNSCC).
Authors: Kang Hyunseok , Huang Jing , Mao Hui , Chen Zhuo Georgia , Peng Xianghong , Shin Dong Moon
Published: March 2020

e13110

Background: Cisplatin (DDP) is a widely used anticancer drug but its application has been limited by side effects and drug resistance. We have previously showed that selective delivery of DDP to tumors using novel nanoparticles could significantly enhance antitumor effects with reduced toxicity. As our group has reported that 45% of 105 HNSCC primary tumors were positive for folate receptor (FR) in Head Neck 31:475-81, 2009, we further developed DDP-loaded folate-targeted iron oxide nanoparticle (FA-IO-DDP) for HNSCC imaging and therapy. Methods: HNSCC cell lines were screened for FR expression by Western blot and surface expression was confirmed by FACS. Intracellular platinum (Pt) and Pt-DNA adducts were quantified by ICP-mass spectrometry. MTT assay was used for cytotoxicity test. T2 weighted MRI was used to detect signal changes induced by FA-IO-DDP. Biodistribution of the nanoparticles was studied using nude mice bearing high FR expressing KB3-1 xenograft. Results: Five cell lines expressing surface FR were identified among sixteen cell lines screened. Three cell lines, PCI15B, SQCCY1 and UMSCC11B with high, intermediate, and negative FR expression respectively, were selected for further investigation. FA-IO-DDP showed significantly enhanced cytotoxicity. Intracellular Pt and Pt-DNA adducts were increased in FR-positive PCI15B cells treated with FA-IO-DDP by 4 fold and 3.2 fold, respectively, compared to cells treated with non-targeted IO-DDP. Similar degree of enhanced uptake was observed in SQCCY1 but not in UMSCC11B. A significant change in T2 signal induced by increased internalization of IO in FA-IO-DDP treated PCI15B cells was detected by MRI. In KB3-1 xenograft model, FA-IO-DDP showed 5.4 fold higher accumulation in tumor compared to free DDP. Conclusions: Our study showed that novel FA-IO-DDP targeting FR remarkably enhanced antitumor effects of DDP in FR-positive HNSCC cells by increasing intracellular Pt concentration and Pt-DNA adducts. MRI may be used to monitor DDP delivery by IO nanoparticles in vivo. Ongoing studies are focused on antitumor effect and toxicities of the FA-IO-DDP using nude mice bearing PCI15B, SQCCY1 and UMSCC11B cells.
Association of clofarabine (Clo)-associated GFR decline (GFRd) with dose and AUC.
Authors: O'Donnell Peter H. , Innocenti Federico , Wickrema Amittha , Stock Wendy , Hardj Marjie , Artz Andrew S. , van Besien Koen
Published: March 2020

e13120^

Background: Clo, a nucleoside analog, is approved for pediatric ALL, but also has activity in AML. We conducted a ph I-II trial of clo-melphalan-alemtuzumab for allo transplant and found grade 3-5 renal toxicity in 16 of 74 patients, despite prolonging infusion time to reduce Cmax (BBMT 2011, Epub). Methods: Clo pharmacokinetics were analyzed in 65 pts (Clo doses 10-40 mg/m2, infusion time 1 or 3 hrs) of 74 pts; baseline GFR estimated by modified MDRD equation. Clo levels were drawn at 1, 1.5, 2, 3, 5, 7, 9 and 24 hrs following the 1st and 3rd (or 4th) dose. GFRd (in ml/min/m2) between start of Clo (d-7) and day of transplant constituted the outcome of interest. Results: Median age 53 (21-73), median baseline GFR 94 ml/min/m2 (46-120). Mean (range) Cmax, clearance (CL) and AUC were 531 ng/ml (119-1010), 42002 ml/min (13979-88020) and 2052 ng*hr/ml (471-5835), respectively. Mean AUC per dose was 32.0 ng*hr/ml/mg (14-32). Systemic exposure was similar between d1 and d3. Further analyses used the highest AUC and Cmax. Decreases in GFR ranging from 10-100 ml/min/m2 (average 44 ml/min/m2) occurred in 30 of 65 patients. There was no correlation between Clo dose and GFRd (r=-0.02,p=0.87). There was a significant correlation between Clo AUC and GFRd (r=0.53, p <0.0001), Clo AUC per dose and GFRd (r=0.62, p<0.001) and between Clo CL and GRFd (r=-0.38, p=0.0001). There was a weaker correlation between Clo Cmax and GFRd (r=0.22, p=0.07). Clo CL was highly correlated with baseline GFR (r=0.4, p=0.0009). Of interest baseline GFR was inversely correlated with pt age (r=-0.46, p=0.0001) and age was significantly correlated with AUC (r=0.4, p=0.001), AUC per dose (r=0.36, p=0.003) and with GFRd (r=0.34, p=0.005). Conclusions: Decline in GFR is associated with inter-patient variability in exposure to Clo. Baseline GFR affects Clo CL and thus AUC, and potentially increases risk for renal toxicity. Older age is associated with decreased baseline GFR and risk for toxicity. Clo may require dose adjustment based upon renal function and/or age. These data require confirmation in further studies.
Targeting the upregulation of the AKT pathway and homologous recombination repair as a therapeutic strategy for epithelial ovarian cancer.
Authors: Ratner Elena , Lin Z-Ping , Rutherford Thomas J. , Azodi Masoud , Santin Alessandro , Schwartz Peter E. , Sartorelli Alan
Published: March 2020

e13125

Background: Epithelial ovarian cancer (EOC) is the second most common gynecologic cancer in the United States, and carries the highest mortality in this category in the West. The progression free survival and overall survival depend greatly on tumor sensitivity to a platinum chemotherapy. Once platinum resistance is encountered, response rates of only 6–30% are achieved. A relatively new modality in EOC that would allow targeted treatments is a PARP inhibitor, a drug that inhibits the enzyme poly (ADP-ribose) polymerase (PARP), which is showing promise for the treatment of EOC with mutations in the BRCA1 or BRCA2 tumor suppressors. Triapine, a novel small-molecule drug developed in our laboratory, potently inhibits the activity of ribonucleotide reductase involved in the key step of DNA synthesis and replication. Methods: 1. Cell sensitivity to varying ratios of treating drug combinations (Triapine with cisplatin; PARP inhibitor, olaparib) was carried out by clonogenic survival assays using multiple EOC cell lines (A2780, Caov-3, EFO, IGROV-1, BG-1, PEO1, SKOV3). 2. AKT level was measured in the cell lines before and after treatment. 3. BRCA1 wild-type and BRCA1-knockdown EOC cells were treated with cisplatin or PARP alone and in combination with Triapine. Drug-induced DNA damage was assessed by the levels of g-H2AX, the marker of double stranded breaks (DSBs), and of Rad51 foci, a marker of HR repair of DSBs. Results: Treatment with Triapine leads to synergistic sensitization of BRCA1 wild-type EOC cells to platinum drugs and to olaparib. Both platinum drugs and olaparib induce DNA damage that is repaired by HR. This suggests that Triapine inhibits HR and sensitizes EOC cells to these drugs. Triapine attenuates olaparib-induced Rad51 foci in BRCA1-wild type cells, which resembles the impairment of such foci formation in BRCA1-knockdown cells. Triapine causes down-regulation of AKT activity in EOC cells. Conclusions: Triapine produces synthetic lethality by inhibiting both DNA repair and pro-survival pathways. Combination of Triapine and platinum drugs/PARP inhibitors represents a rational and innovative therapy that targets EOC with a high level of AKT activity.
ApoStream, a novel device for antibody-independent capture of circulating cancercells (CCCs) from blood of patients with various types of cancer.
Authors: Gupta Vishal , Jafferji Insiya , Garza Miguel , Sukumaran Sujita , Woo Jacky , Hasegawa Dave , Melnikova Vladislava O. , Davis Darren W.
Published: March 2020

e13500

Background: The detection of circulating tumor cells (CTCs) using immunomagnetic EpCAM-based capture methods has been conceptually accepted as a “liquid tumor biopsy”. However, these methods have limited the recovery of CTCs for molecular profiling applications. We developed a novel continuous flow dielectrophoresis field-flow fractionation (DEP-FFF) device, ApoStream for antibody-independent capture of circulating cancer cells (CCCs), with improved recovery across multiple cancer types and preserved viability of CCCs for downstream characterization. Methods: The performance of ApoStream was demonstrated using a low EpCAM expressing cell line, SKOV3. ApoStream was further used to enrich CCCs from various cancer patient blood. Prostate, breast and NSCLC CCCs were stained for cytokeratin (CK), CD45, and DAPI; melanoma CCCs were stained with S100, CD45 and DAPI. CCC enumeration was performed using laser scanning cytometry. Results: In system precision performance studies, average inter-day recovery on the ApoStream was 80.3 ± 3.5%, CV = 4.3% when cancer cells were spiked into buffer, and 78.5 ± 3.0%, CV = 3.3% when cancer cells spiked into ~10 million healthy peripheral blood mononuclear cells (PBMCs). Linearity performance was demonstrated with spiking 5-2600 SKOV3 cells into 10 million PBMCs (R2=1). Cell viability was not affected by processing through ApoStream device. High CCC recovery from metastatic cancer patient blood samples was obtained with counts ranging from 0 - 2630 (NSCLC, n=66), 0 - 3490 (prostate, n=29), 10 - 968 (breast, n=11), and 3 - 3120 (melanoma, n=13) CCCs per 7.5 mL blood. Positive CCC counts were obtained in 90% of NSCLC samples, 93% of prostate cancer samples, 100% breast cancer and melanoma specimens. There were no CK+ cells detected in healthy donor blood controls. Conclusions: Improved CCC recovery from various cancer types was demonstrated with the ApoStream device. ApoStream provides an antibody-independent method for capture of viable CCCs that enables further downstream molecular characterization of rare cells for use in clinical applications. Acknowledgements: Funded by NCI Contract No. HHSN261200800001E.
Effect of microRNA-124 on homologous recombination protein RAD51 and glioblastoma cells undergoing radiation.
Authors: Schäfer Henning S , Agami Reuven , van Haaften Gijs
Published: March 2020

e13515

Background: The DNA damage repair (DDR) pathways are an essential cell survival mechanism to preserve genomic integrity. In tumor cells, DDR pathways have a huge impact in cell survival, response to therapy and thus prognosis. Our objective is to investigate whether microRNAs (miRs) can influence specific DNA repair pathways. For our studies we focussed on double-strand break (DSB) repair by homologous - (HR) or non-homologous recombination (NHEJ). DSBs are the main lethal lesions induced by ionizing radiation. Methods: A retroviral miR expression library of ~500 miRs was transduced into a HR reporter cell line. For less abundant miRs found in this analysis in silico analysis was performed to detect potential targets. Regulation of HR protein RAD51 by miR-124 could be shown by luciferase assays and immunoblotting. Overexpression of miR-124 in glioblastoma cell line U87 was done by lentiviral constructs. Effect of miR-124 overexpression was revealed by colony – and competetive growth assays. Results: In an unbiased screen we found that miR-124 is less abundantly expressed in a reporter cell population sufficient for HR, suggesting a negative role for these miRs in DSB repair via HR. Reporter assays revealed a negative regulation of RAD51, a key protein of HR pathway, by miR-124. This regulation was specific to a conserved binding motif at the 3` UTR of RAD51. This regulation could also be detected in vivo whereas overexpression of miR-124 leads to downregulation of RAD51 protein levels. Functional experiments in glioblastoma cell line (U87), that express hardly no miR-124, revealed increased sensitivity to ionizing radiation if miR-124 is (re)-expressed. Conclusions: In this work we uncover miRs influencing the cellular DNA-repair machinery. We show that miR-124 influences the repair of DSBs by HR. This effect is specific and mediated by the regulation of RAD51 protein level, an essential component of the HR pathway. Re-expression of miR-124 sensitizes glioblastoma cell line (U87) to radiation in functional experiments. We propose that (re)-expressing miR-124 in glioblastomas could be a new treatment strategy to sensitize this tumors to radiation therapy.
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.