bluuebunny/crossref_metadata_2025_split huggingface dataset:
Relation between lactate dehydrogenase (LDH) during two first cycles in patients with metastatic renal cell carcinoma (mRCC) treated with sunitinib (SU)
Authors: Medioni J. , Arakelian N. , Fournier L. , Helley D. , Thiam R. , Banu E. , Cuenod C. , Oudard S. 
Published: March 2020

14079 Background: LDH is a biomarker of cellular turn over. We evaluated LDH as a surrogate marker of tumor response and tumor necrosis during first cycles of SU. Methods: This is a single center study of immunorefractory patients with mRCC treated in second line with SU (50 mg, 4 weeks on, 2 weeks off). All patients performed CT scan and had serially measured tumor necrosis. Tumor response using the RECIST criteria was evaluated at the end of cycle 2 (C2). Tumor necrosis index was calculated at baseline, at the end of cycle 1 (C1) and C2 by measuring the percent area of the tumor with no enhancement after injection using an electronic calliper. LDH was measured at baseline (D1) and at the end (D28) of C1 and C2 using LD 200 kit on Synchron LX20 (Beckman Coulter). Relations between LDH and tumor response were studied using multivariate analysis of variance (MANOVA) with repeated measures and between LDH and tumor necrosis using Pearson correlation test. Results: Between Feb 2005 and Aug 2006, 56 patients were analyzed (75% men,), 91% Clear Cell Carcinoma. Mean age was 59 years (range: 30–81) and 62% has a performance status ECOG of 0. Median number of metastatic sites was 2 and mean hemoglobin level was 12.7 gr/dl. After C2, 12 (21%) partial responses, 35 (63%) stable diseases and 9 (16%) progressive diseases were noted. Results of LDH are summarized in the table . Mean LDH was statistically different between along time (p < 0.0001). LDH levels were statistically different between responders-stables and progressors patients (p = 0.004). There is a trend for a statistical relation between the index of necrosis and the tumor response measured by RECIST criteria (p = 0.08). No correlation was found between LDH and tumor necrosis. Conclusions: LDH level is higher in progressors compared to responders-stables patients. LDH level is not correlated to tumor necrosis measured by CT scan. [Table: see text] No significant financial relationships to disclose.
CD40 ligand gene therapy directly induces apoptosis in carcinomas
Authors: Palmer D. H. , El-Metwali T. , Milner A. E. , Hodgkin E. , Young L. S. 
Published: March 2020

14082 Background: CD40 is a member of the TNFReceptor superfamily expressed on a variety of immune cells. Interaction of CD40 with its ligand (CD40L) plays a key role in orchestrating immune responses. We have shown that CD40 is also expressed on a variety of carcinomas. CD40 ligation in this context can induce cancer cell death. Thus CD40 represents a rational target with direct cytotoxicity and induction of anti-tumour immunity. Methods: We generated an adenovirus encoding CD40L (AdCD40L) and investigated its effect in CD40-expressing carcinoma cells (bladder, ovary, hepatocellular) as follows:(i)comparison of Ad-delivered CD40L with recombinant soluble ligand (rsCD40L);(ii)inhibition of cleavage of CD40L from the cell membrane using metalloproteinase inhibitors (MMPI);(iii)generation of a mutant CD40L resistant to MMP cleavage. Results: (i)CD40 ligation in carcinomas stimulates survival and apoptotic signaling. rsCD40L is only cytotoxic in the presence of protein synthesis inhibition, which blocks survival pathways and allows apoptosis. In contrast, AdCD40L is cytotoxic even in the absence of protein synthesis inhibitor;(ii)AdCD40L results in expression at the cell membrane but this is cleaved from the membrane and detected as soluble ligand in supernatant of infected cells. Cleavage is inhibited by MMPI and this increases apoptosis, suggesting that membrane-bound CD40L is a more potent apoptotic stimulus than soluble ligand;(iii)we identified the cleavage site of CD40L and generated a mutant by deletion of this region and cloned this mCD40L into an adenovirus. AdmCD40L results in membrane expression of CD40L, with no detectable soluble CD40L, confirming its resistance to cleavage. AdmCD40L is significantly more cytotoxic than wild-type AdCD40L. Conclusions: The effect of CD40 ligation depends on the cellular context and the method of CD40L delivery: adenovirus delivery results in membrane-bound expression and this provides a stronger apoptotic stimulus than rsCD40L. Apoptosis is further enhanced by a mutant CD40L that is resistant to cleavage from the cell membrane. Thus AdmCD40L is a novel therapeutic that is a potent inducer of apoptosis. Studies are underway to assess these effects in vivo and to elucidate the contribution of immunostimulatory effects in syngeneic models. No significant financial relationships to disclose.
Measurement of gene amplification in FFPE tumor tissues by quantitative real-time PCR in comparison with FISH
Authors: Boyer S. , Yang J. 
Published: March 2020

14086 Background: Amplified oncogenes such as HER2 and EGFR have proven to be good targets for molecular therapy. Recent retrospective analyses suggest that EGFR amplification predicate better responses to EGFR kinase inhibitors. However, different methods were used to measure EGFR gene amplification and polysomy in these studies, including FISH and various real-time quantitative PCR platforms, which may have led to the different conclusions reported by these groups. In this study, we compared Taqman- QPCR with FISH in scoring EGFR and HER2 amplification and EGFR polysomy on 42 FFPE tumor samples. Methods: DNA was prepared from 42 FFPE tumor tissues of breast, colon, lung, and stomach origin. Taqman QPCR was carried out according to manufacturer’s protocol on an ABI-7900HT instrument. FISH analysis for EGFR and HER2 amplification was carried out according to Vysis standard protocols. Results: QPCR reference probes for measuring EGFR and HER2 amplification were designed against DNA fragments near the centromeric regions of chromosome 7 and chromosome 17, respectively. A QPCR probe for RNase P on chromosome 6 was designed and used as a reference for measuring EGFR polysomy. HER2 amplification was detected in ∼25% FFPE breast tumor samples by both FISH and QPCR methods, although amplification scores were consistently higher by QPCR than by FISH. EGFR amplification was rare in all samples screened but high polysomy (6 copy and above) was detected in 40% FFPE gastric tumor samples. Again there was high concordance between FISH and Taqman QPCR results. Conclusions: Our data showed high concordance between Taqman QPCR and FISH in scoring EGFR and HER2 amplification. Taqman QPCR is a highly sensitive, accurate, convenient, and economical technology for measuring EGFR and HER2 amplification and EGFR polysomy in FFPE tissues. With well validated target and reference primers/probes, Taqman QPCR provides an attractive alternative to FISH for measuring oncogene amplification for retrospective studies on archived FFPE samples and for screening patients for clinical trials. No significant financial relationships to disclose.
Hepatic expression of UGT enzymes and implications in pharmacokinetic variations in Asians during chemotherapy
Authors: Chowbay B. , Lal S. , Shu X. , Jada S. , Ooi L. , Liu Y. , Ong Y. 
Published: March 2020

14089 Background: The objective of this study was to screen the promoter and relevant coding regions in the UGT1A1 and UGT1A9 genes and genotypically quantitate expression of these genes in hepatic tissues obtained from Asian cancer patients undergoing hepatectomy. Methods: Non-cancerous Asian hepatic tissues (Chinese: N=36; Malay: N=1; Indian: N=3) were obtained from the institution’s central tissue repository bank. Messenger RNA was isolated from each tissue sample and reverse transcribed to cDNA followed by real time PCR experiments. Results from these gene expression experiments were normalized against GAPDH transcripts. DNA was also extracted from the hepatocytes, screened for variations in the promoter regions (UGT1A1*60, g.3140G>A and UGT1A1*28) and exon 1 (UGT1A1*6) of the UGT1A1 and UGT1A9 (UGT1A9*22, I399C>T) genes. Results: Large interindividual variations were observed in the hepatic expression pattern of UGT1A1 and UGT1A9 mRNA with respect to the different polymorphic variants. There was no significant association between hepatic expression of UGT1A1 mRNA and UGT1A1*60, g-3140G>A and *6 genotypes (P>0.05 in each case). The median expression level of UGT1A1 mRNA was approximately 2-fold higher in liver tissues harboring UGT1A1 6/6 (reference) genotype (N=27, median = 0.478; range: 0.065 to 3.125) compared to the heterozygous 6/7 genotype (N=9, median = 0.254; range: 0.042 to 1.894; P=0.77). The median expression levels of UGT1A9 mRNA were approximately 3-fold higher in liver tissues harboring I399CC genotype (N=3, median = 2.735; range: 0.702 to 3.375) when compared with the homozygous variant TT genotype (N=8, median = 0.973; range: 0.652 to 4.155; P=0.88). Conclusions: The results showed that polymorphisms in the UGT1A genes may influence the expression pattern of the respective proteins and could contribute to the high degree of interpatient variability in the glucuronidation dependent metabolism and elimination of chemotherapeutic agents in Asian cancer patients. No significant financial relationships to disclose.
Pediatric phase I trial and pharmacokinetic (PK) study of dasatinib: A report from the Children’s Oncology Group
Authors: Aplenc R. , Strauss L. C. , Shusterman S. , Ingle A. M. , Luo R. , Wright J. , Blaney S. , Adamson P. C. 
Published: March 2020

14094 Background: Dasatinib is an orally-available tyrosine kinase inhibitor with low-nanomolar activity against SRC-family kinases, BCR-ABL, c-KIT, EPHA2, and the PDGFβ receptor. Methods: A phase I study of dasatinib administered as a single agent in pediatric patients with refractory solid tumors or imatinib refractory Ph+ leukemias is being performed. Dasatinib dose levels of 50, 65, 85 and 110 mg/m2/dose, administered orally twice daily for 28 consecutive days, are being studied. Courses repeat without interruption. Results: 8 pts with solid tumor and 3 pts with Ph+ leukemia (median age 11 yrs, range 7–17) have been enrolled, of whom 7 are fully evaluable for toxicity. At the 50 and 65 mg/m2 dose levels, 0/3 and 0/1 pts experienced DLT. 1 of 3 pts with CML treated at the 50 mg/m2 dose level had Gr 4 hypokalemia. In 4 pts studied at the 50 mg/m2 dose level, the median (range) apparent dasatinib clearance was 238.5 L/hr (220.7 - 332.7), terminal half-life was 2.3 hrs (2.2 - 3.1), and the median Cmax was 55.5 ng/ml (36.0 - 161.1). Conclusions: Dasatinib is well tolerated at a dose of 50 mg/m2 BID on a continuous 28 day dose schedule in pediatric patients. Preliminary PK analysis suggests that drug disposition is similar to that observed in adults. Dose escalation and PK studies are continuing. No significant financial relationships to disclose.
Dual AKT inhibition with perifosine and GCP enhances apoptosis in prostate cancer: Clinical implications
Authors: Vinall R. L. , Hwa K. , Pan C. , Ghosh P. , Lara P. N. , deVere White R. W. 
Published: March 2020

14096 Background: Genistein combined polysaccharide (GCP) has been shown to exhibit anti-cancer properties in both experimental and clinical models of prostate cancer (CaP). Perifosine is an alkylphospholipid with clinical anti-neoplastic activity. Both agents inhibit the AKT signaling pathway. Methods: Prostate cancer cell lines (LNCaP, LNCaP stably transfected with R273H or P151S p53 mutant allele, cds1, and PC3) were treated with GCP, perifosine, or both. IC50 values were established using the MTT assay. Apoptosis was assessed by flow cytometry, Western blot of PARP cleavage, and caspase activity. Clonogenic potential was assessed by colony assay. Status of AKT, p53, p21, AR and PSA was determined by Western blot. All experiments were performed in triplicate. Results: Perifosine inhibited AKT activity in all the cell lines. GCP had little or no effect on AKT activity but reduced AR and PSA levels. The combination of GCP and perifosine further increased the level of AKT inhibition and maintained inhibition for longer when compared to treatment with perifosine alone. Flow cytometric analysis of LNCaP revealed that combination treatment dramatically increased SubG1 levels (23-fold increase versus a 4.4-fold increase for GCP alone and a 6.5% increase for perifosine alone). Apoptosis was confirmed by PARP and caspase analysis. As single agents, the main effect of GCP or perifosine was to induce growth arrest as shown by a decrease in S-phase and increased p21. The cds1 cell line responded similarly to LNCaP, however, cell lines that expressed mutant p53 or were p53 null were not susceptible to GCP/perifosine-induced apoptosis. Combination treatment further decreased the clonogenic potential in all of the cell lines assessed when compared to treatment with the single agents alone. Conclusions: Treatment with a combination of the AKT inhibitors GCP and perifosine dramatically increases apoptosis and/or inhibits clonogenic potential in several CaP cell lines. The effect of the combination treatment on apoptosis appears to be dependent on p53 status. Clinical validation of these findings is warranted. A clinical trial of hormone therapy with or without GCP/perifosine is presently in development. No significant financial relationships to disclose.
Recruitment experience in a phase 0 trial of ABT-888, an inhibitor of poly (ADP-ribose) polymerase (PARP), in patients (pts) with advanced malignancies
Authors: Gutierrez M. , Kummar S. , Horneffer Y. , Juwara L. , Chen A. , Melillo G. , Pickeral O. , Tomaszewski J. E. , Murgo A. J. , Doroshow J. H. 
Published: March 2020

14111 Background: Phase 0 trials conducted under the FDA Exploratory IND Guidance involve limited exposures to study drug with no expectation of clinical benefit. We present our experience in recruiting pts to a Phase 0 study of ABT-888, the first to be conducted at the NCI. Methods: Objective: To determine the ABT-888 dose that inhibits PARP in PBMCs and tumor after a single oral administration. Tumor biopsies at baseline and 3–6 hrs post drug were planned once PARP activity inhibition in PBMCs or target plasma Cmax was achieved. Protocol was approved by the NCI scientific committee, IRB, and CTEP, and was reviewed by the NIH Ethics Committee. Informed consent included an explicit statement acknowledging the non-therapeutic, non-personal benefit nature of the study. Results: Since 6/06, 17 pts screened; 6 enrolled. Reasons for declining participation: Non-therapeutic study (2); biopsy requirement (2); recommendation by oncologist (2) or family member (2); condition requiring treatment (3). Reasons for participation: Altruism (4); altruism + waiting for another study (2). Of 6 pts enrolled, colon cancer (2), lymphoma (2), carcinoid (1), lung cancer (1). Median age: Participants (P) 57 (49–64) and non-participants (NP): 62 (42–78). Median prior therapies: P 3.6 (0–8) and NP: 3 (0–7). Referral origin for P: Prior participation on NCI studies (2), prior pt-NCI physician relationship (4). Ethnicity of P: White (5) and Hispanic (1). 3 of 6 pts underwent serial biopsies as required by protocol. Conclusion: Phase 0 recruitment is feasible and the accrual can be completed in a timely fashion, but depends on pt altruism and prior physician-pt relationship. These studies can be ethically conducted provided that interventions are of minimal risk, the study experience is commensurate with pt expectations, and that results are invaluable to subsequent pts. No significant financial relationships to disclose.
Predicting and monitoring tumor response to epidermal growth factor receptor inhibitor gefitinib in patients with locally advanced esophageal adenocarcinoma
Authors: Gibson M. K. , Mezzadra H. , Kleinberg L. , Jagannath S. , Brock M. , Abdallah N. , Rudek M. , Berman D. , Forastiere A. , Altiok S. 
Published: March 2020

14112 Background: This study aimed to validate an ex vivo chemosensitivity assay to measure the pharmacodynamic effect of gefitinib on esophageal adenocarcinoma (EAC) prior to treatment with pre-operative concomitant chemoradiotherapy (CRT). Methods: A 14 day run-in period with 250 mg/day of gefitinib preceded CRT. Endoscopic biopsies (D 0 and 14) in 4 patients with T2–3N0/1M0/1a EAC were analyzed by ex vivo chemosensitivity assay. Day 0 tissue was exposed to gefitinib ex vivo, then tumor was exposed to gefitinib for 14 days in vivo (ie in the patient). Phosphorylation of the EGFR, raf/MEK/ERK and PI3/AKT pathways was measured by Western blot. Profiles were compared for correlation between ex vivo and in vivo exposure, and patterns were correlated with response to CRT. The effects were also characterized by immunohistochemistry (IHC). EGFR, K-Ras, and PI3K mutations, serum concentrations of gefitinib and PTEN status were measured as potential confounders. Results: One patient with stage T3N1 died of unexplained hemorrhage during surgery. Three had clinical and path stages of: T3N1/T0N0, T3N0/T3N0, T3N1/T2N1. Gefitinib levels were constant, confirming exposure of target tissue to the drug. Ex vivo exposure yielded four distinct pathway patterns. The exact same patterns were seen after in vivo exposure. No mutations were identified in exons 18–21 of the EGFR, exons 2/3 of K-ras or exons 9/22 of PI3K. PTEN levels were similar in all tumors. PCNA expression correlated with raf/MEK/ERK pathway inhibition, but not with inhibition of EGFR activity. IHC correlated with Western blot for expression of EGFR, and phospho- and total ERK levels. No correlation was observed between gefitinib effect and pathologic response to CRT. Conclusions: This study used a novel ex vivo chemosensitivity assay to demonstrate the activity of gefitinib to inhibit target in tumor tissue obtained from patients with EAC. The exact correlation of pre- and post-treatment profiles suggests potential use in the pre-treatment setting to predict in vivo effects of targeted therapies. This approach may facilitate the further refinement of patient selection to maximize potential benefit while sparing patients unlikely to respond to a given agent. No significant financial relationships to disclose.
Hypertensison is a significant adverse effect of bevacizumab treatment
Authors: Lowery M. A. , Power D. G. , Behbehani A. T. , Carney D. N. , McCaffrey J. A. 
Published: March 2020

14134 Introduction: Vascular endothelial growth factor (VEGF) is upregulated in many tumours. Bevacizumab (BV) is a recombinant, humanised monoclonal antibody against the VEGF receptor and has shown promising results in phase II / III trials in colorectal cancer, non-small cell lung cancer, renal cell carcinoma, and breast cancer. Hypertension (HT) has been reported in all studies involving BV and management of this is not well established. Methods: The records of patients treated with BV in our institution were analyzed from January to August 2006. Indications for BV therapy, incidence and management of HT, and resulting outcomes were assessed. Results: 71 patients (45 male, 28–84 years) were treated with BV, at a dose of 5mg/kg every two weeks. 87% (n=60) had colon cancer. Lung cancer, renal cell carcinoma, melanoma, and pancreatic cancer accounted for the rest. 20% (n=14) had pre-existing HT. In all of these cases HT was well controlled with medication. 10 patients developed new-onset HT, and 7 patients experienced exacerbation of existing HT. Using the CTCAE(v3.0) guidelines for grading of HT, 5 had grade II and 10 had grade III. HT developed after a median of 15 weeks of BV (range 6–36 weeks), and a median cumulative BV dose of 20mg/kg (range 10 - 85 mg/kg). In the new onset HT cases 8 patients were controlled (BP= 140/90) with a single anti-HT, and one patient needed two or more agents (BP<160/100). In the pre-existing HT cases who needed an adjustment of anti-hypertensives, 3 were controlled (BP=160/100) with two or more agents and 4 patients required dose escalation of a single agent. In 2 patients BV was permanently discontinued. Anti-HT agents used included beta-blockers, ACE-inhibitors and Ca++- channel blockers. Conclusions: HT may develop at any point during treatment with BV. BP should be measured regularly. In this study 14% of patients developed HT de novo, and 50% experienced exacerbation of pre-existing HT. Median time to BV-induced HT was 15 weeks. There is currently no data to define which anti-HT is the best to use. BV is now becoming widely used, and as with all new targeted therapies, it is important that oncologists recognise relatively unusual side effects, and develop appropriate practice guidelines. No significant financial relationships to disclose.
Pre-clinical evidence for an anti-tumor activity of defibrotide, a DNA-based, endothelium stabilizing drug
Authors: Echart C. , Iacobelli M. , Richardson P. , Mitsiades C. , Ignoni T. , Ferro L. , Eissner G. 
Published: March 2020

14144 Background: Defibrotide (DF) is a mixture of polydeoxyribonucleotides with anti-thrombotic activity. Next to endothelium stabilization, recent data suggest anti-neoplastic properties of DF modulating interactions of tumor cells with their microenvironment. We investigated whether DF regulates expression and activity of heparanase, an enzyme critically involved in breaking down extracellular barriers and releasing growth factors linked to tumor invasion and angiogenesis. Methods: Heparanase expression was tested by RT-PCR and flow cytometry with multiple myeloma (MM) and microvascular endothelial cells. Heparanase activity was measured in cellular extracts with a heparan-degrading enzymatic assay. Serum degradation products of DF were identified by SEC-HPLC. The anti-angiogenic potential of DF was tested in vitro using a kit with human microvascular endothelial cells forming tubes across a layer of fibroblasts. In vivo, DF was tested in the dorsal skin-fold chamber assay in mice after inoculation of human gastric cancer cells. Proliferation was assessed by trypan blue exclusion. Results: We demonstrate a striking downregulation of expression and enzymatic activity of heparanase in endothelial as well as MM cells. In contrast, the degradation products of DF failed to exert any biological activity, suggesting that the intact mixture of deoxyoligonucleotides is responsible for the anti-tumor effect. We could also show that DF prevents (tumor) angiogenesis in vitro and in vivo. Western blots suggest that DF reduces phosphorylation-activation of p70S6 kinase, a key target in the mTOR pathway linked to angiogenesis. In addition, DF does not influence proliferation of vascular or tumor cells, rather acts via selective inhibition of tube formation of endothelial cells. Conclusion: In the present report we provide evidence for an anti-tumor activity of DF. DF inhibits (tumor) vessel formation and heparanase activity, and thus should be considered as an anti-cancer agent. [Table: see text]