bluuebunny/crossref_metadata_2025 huggingface dataset:
Pretherapy predictive immunomodulation of NK cell activity and expression of activating and inhibitory receptors in stage IV melanoma patients
Authors: Konjevic G. , Mirjacic K. , Radovanovic A. , Jovic V. , Babovic N. , Spuzic I.
Published: March 2020

12514

Background: As melanoma (MM) is an immunogenic tumor with poor response to chemotherapy immunomodulating agents are applied in order to potentiate cytotoxic effect of chemotherapy and enhance antitumor immune response. Beside therapeutic benefit of IFN-α and IL-2, 13-cis retinoic acid (RA), as an antiproliferative, differentiating and immunomodulating agent is also investigated. The effect of these agents on NK cells, as main innate immune system effectors, is being investigated. Methods: 35 patients with MM in stage IV prior to therapy and 20 controls were investigated. We evaluated NK activity and the expression of activating (NKG2D and CD161) and inhibitory (CD158a and CD158b) receptors on freshly isolated PBL. Predicitive immunomodulation was performed in 18 h in vitro treated PBL with rh IL-2 (200U/ml), IFN (250U/ml), RA (10−6M), and their combination. Results: Native NK cell cytotoxic activity and expression of NKG2D and CD161 activating receptors on fresh NK cells in MM patients is significantly decreased compared to controls. Predictive treatments with IL-2, IFN, IFN and RA, unlike RA alone, gave a significant increase in NK cell activity of MM patients. Singly, IFN also induced a significant increase in CD161 expression on NK cells in patients. The treatments gave no change in the expression of CD158b, while RA, alone, induced significant decrease in the expression of the inhibitory CD158a antigen. Evaluation of mRNA of transcription molecule IRF-1 shows that it is promptly up-regulated by IFNα, more by IFNα and RA, while single RA has no effect on mRNA induction. Conclusions: Considering that the mechanism of applied immunomodulating agents is continually investigated in order to optimize the dose and schedule of their administration and, also, considering controversial clinical results of RA application, alone, or with IFN, in this study we give novel results that show no significant effect of RA, whereas, IFN-induced increase in NK cell activity of MM patients is for the first time associated with the increase in the expression of NKG2D and CD161 receptors on CD16+NK cells, and not with the decrease in some inhibitory receptors, as the activity of NK cells is regulated by the balance of these two types of signals.

No significant financial relationships to disclose.
Phase I experience with an anti-glycotope monoclonal antibody, RAV12, in recurrent adenocarcinoma
Authors: Burris H. A. , Rosen L. S. , Lewis N. , Jones S. F. , Van Vugt A. , Cohen R. B. , Kunkel L. A. , Stewart S. J.
Published: March 2020

13002

Background: RAV12 is a high affinity, internalizing, chimeric IgG1 monoclonal antibody (MAb) that binds RAAG12, a novel primate-restricted N-linked carbohydrate epitope present on multiple cell-surface proteins. RAAG12 is variably expressed on normal non-keratinizing epithelia and is not expressed on human tissues from the cardiovascular, endocrine, hematolymphatic, neuromuscular, and central nervous systems. The majority of RAAG12 expression is in the cytoplasm. In some cases, e.g. polarized epithelia such as colon and stomach, RAAG12 is located in both the cytoplasm and the apical membrane. IHC studies demonstrated that > 90% of human colon, gastric, and pancreatic adenocarcinoma samples bound RAV12, indicating expression of RAAG12. RAAG12 is distinct from previously reported mucin or Lewis blood group carcinoma-associated carbohydrates. Methods: In December 2004, a phase I dose-escalation safety and pharmacokinetics (PK) trial was begun in patients with recurrent (1 to 3 prior treatments) adenocarcinoma. RAV12 was administered weekly × 4. Fourteen patients have received treatment, 6 in a first cohort at 0.3 mg/kg, and 8 in a second cohort at 1.0 mg/kg. Nine patients had colorectal, 2 gastroesophageal, 2 pancreatic, and 1 lung cancer. Responses were evaluated on day 42. Results: All patients, except one who experienced an HSR during his fourth dose, completed the planned weekly treatments. Three clinical syndromes have been observed to date: 1) abdominal cramping pain and diarrhea (11 patients) particularly at the higher dose, 2) asymptomatic, self-limited, rapidly reversible excursions of liver function tests (3 patients), and 3) asymptomatic, self-limited, rapidly reversible excursions of pancreatic enzymes (2 patients). Three patients had stable disease as a best response at day 43. One patient, with pancreatic cancer, had a > 50% reduction in CA19–9, continued treatment for a total of 9 doses, and had a period of disease stability of 5 months. Preliminary analysis demonstrated dose-dependent PK. Dose escalation continues at 1.5 mg/kg. Conclusions: RAV12 may have activity in recurrent adenocarcinoma. Side-effects, though present, have been manageable. Further experience with the MAb will be required to evaluate safety and activity.

[Table: see text]
A phase II study of gefitinib as a first-line therapy for advanced non-small cell lung cancers with epidermal growth factor receptor (EGFR) gene mutations
Authors: Asahina H. , Yamazaki K. , Kinoshita I. , Ogura S. , Ishida T. , Dosaka-Akita H. , Isobe H. , Nishimura M.
Published: March 2020

13014

Background: Activating mutations in exon 18–21 of the epidermal growth factor receptor (EGFR) gene have been reported to be a predictor of response to gefitinib. We conducted a phase II study to evaluate the efficacy and safety of gefitinib as a first-line therapy for advanced non-small cell lung cancers (NSCLCs) with EGFR gene mutations. Methods: The primary endpoint was response rate (RR). Eligiblity criteria were stage IIIB or IV chemotherapy naive NSCLC, ECOG PS of 0–2, adequate organ functions, measurable lesions, and written informed consent. First, we examined EGFR gene mutations in exon 18–21 by direct sequencing of PCR products of DNA extracted from paraffin-embedded tissues. Those who had EGFR gene mutations received gefitinib 250 mg daily. This study, with a sample size of 14, had 90% power to support the hypothesis that true RR was ≥70%, and 5% significance to deny the hypothesis that true RR was ≤30%. Assuming an inevaluality rate ≤15%, we projected an accrual of 16 patients. Results: From Nov. 2004 to Jan. 2006, EGFR gene mutations were analyzed in tumor specimens from 82 patients. Twenty patients (24%) had EGFR gene mutations (16 with deletions in or near E746-A750, 4 with L858R). While there were no significant differences between mutation status and age, sex, histology, or the procedures to obtain tumor specimens, EGFR gene mutations were more frequently observed in never-smokers than in smokers (39% vs. 11%, p<0.01). Sixteen patients (median age, 68; male/female, 3/13; adenocarcinoma/SCC, 15/1; current/former/never smoker, 2/1/13) were enrolled and treated with gefitinib. To date, best response is 2 CR, 7 PR, 1 SD and 1 PD (RR, 82%). Two patients had grade 3 toxicities, including 1 skin eruption and 1 liver dysfunction. One patient had grade 1 interstitial lung disease, leading to the termination of gefitinib treatment. Conclusions: Gefitinib is very active and well tolerated as a first-line therapy for advanced NSCLCs with EGFR gene mutations.

No significant financial relationships to disclose.
Rapid detection of UGT1A1 polymorphisms by a novel microarray assay: A validation study using 100 samples from patients with lung cancer treated by irinotecan
Authors: Yamada Y. , Shimizu T. , Ozaki T. , Hatashita E. , Kurata T. , Fukuoka M. , Tamura K.
Published: March 2020

13019

Background: Irinotecan is one of the active drugs for the treatment of colorectal, gastric and lung cancer. SN-38, the active metabolite of irinotecan is metabolized through conjugation by uridine diphosphate glucuronosyl transferase (UGT1A1). Genetic polymorphisms of UGT1A1 in the specific locus have been reported to be significantly related to severe toxicities of Irinotecan. Although the polymorphisms are usually detected by direct sequence (DS) or PCR-restriction fragment length polymorphism (RFLP) analysis, these assays consume time and need deferent PCR product for each exon. We report a novel microarray (MA) technology to detect the UGT1A1 polymorphisms using 100 peripheral blood samples of lung cancer patients treated with irinotecan, and evaluate a consistence ratio in comparison with the confirmed results by RFLP or DS. Methods: One hundred lung cancer patients who had received irinotecan containing chemotherapy were enrolled in this study. Genomic DNA was extracted from whole blood and prepared into 100 ng/5μL. PCR were performed by random primers, and the PCR products were labeled by biotin. The labeled probe have hybridized the MA chips, and patterns of visualized dots informed us the polymorphisms in the deferent locus (*28, *6 *27), simultaneously. Polymorphisms of all samples were also confirmed by RFLP and DS for validation. Results: One homozygous (Hom), 14 heterozygous (Het) and 85 wild type (W) for UGT1A1*28 were confirmed by DS. Five Hom, 25 Het and 70 W for UGT1A1*6, and 1 Het and 99 W for UGT1A1*27 were confirmed by RFLP and DS. A single approach of the novel MA assay have revealed 79.3%, 96.6% and 100% consistence with the confirmed result for each exon (*28, *6 *27), respectively. Individual twice approach of the MA have revealed 94.3%, 98.9% and 100% consistence with the confirmed result for the each exon, respectively. We could assay all samples by the MA and needed only single day for detection. Conclusions: Our newly developed assay for detecting UGT1A1 polymorphisms is rapid, accurate and feasible and has potential to be used for screening before irinotecan in the future prospective clinical trials.

No significant financial relationships to disclose.
[18F]fluorothymidine (FLT) PET after 3 days of gefitinib treatment and tumor response in non-small cell lung cancer (NSCLC)
Authors: Sohn H. , Kim S. , Ryu J. , Oh S. , Moon D. , Oh Y. , Shim T. , Kim W. , Suh C. , Lee J.
Published: March 2020

13031

Background: FLT has been developed as a PET tracer for imaging tumor proliferation. We evaluated whether FLT-PET could predict tumor response only after 3 days of gefitinib treatment. Methods: Nonsmokers with adenocarcinoma of the lung were eligible for this study. FLT-PET was performed at 1 day before and 3 days after the start of gefitinib (250 mg/d) therapy. The maximum standardized uptake value (SUVmax) of the main lung mass was measured, and changes in tumor SUVmax were calculated. After 6 weeks of therapy, response was assessed by chest CT according to WHO criteria. The cutoff value predicting subsequent CT response was obtained by receiver operating characteristic curve analysis. Results: Between Jun. 2005 and Nov. 2005, 22 patients were enrolled. CT response was partial response in 12 (54%), stable disease in 5 (23%), and progressive disease in 5 (23%). As early as 3 days after the initiation of therapy, significant difference in % changes of tumor SUVmax on FLT-PET was observed between responders and nonresponders (−32% v −2.3%, P = .002) ( Table ). When a reduction of tumor SUVmax ≥ 20% was used as a cutoff value for FLT-PET response, CT response could be predicted with positive and negative predictive values of 100% and 83%, respectively. Time to progression was significantly longer in FLT-PET responders than nonresponders (median 5.1 v 1.4 months, P = .011). Conclusions: Using FLT-PET obtained on days 0 and 3 of gefitinib therapy, the response could be early predicted in patients with NSCLC.

[Table: see text]

No significant financial relationships to disclose.
Insulin-like growth factor-1 receptor/epidermal growth factor receptor (EGFR) heterodimerization and resistance to epidermal growth factor receptor tyrosine kinase inhibitors in non-small-cell lung cancer
Authors: Morgillo F. , Hong W. K. , Lee H.
Published: March 2020

13032

Background: Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) have been used to treat lung cancers, but resistance to these agents has been observed. This study was designed to investigate whether the insulin-like growth factor (IGF)-mediated signaling pathway induces resistance to the EGFR TKIs in lung cancer. Methods: The antitumor activities and action mechanisms of EGFR inhibitors (erlotinib, gefinitib, cetuximab), single or in combination with IGF-IR inhibitors, were assessed in vitro in a subset of non-small-cell lung cancer (NSCLC) cell lines by the MTT assay, flow cytometry-based TUNEL assay, soft agar, confocal microscopy, metabolic labeling, coimmunoprecipitation, and northern and western blot analyses, and in vivo in animal models. Correlation of EGFR and IGF-1R expression was assessed using lung tissues from patients with NSCLC. Results: EGFR TKI inhibited the proliferation and anchorage-dependent and -independent colony-forming ability of NSCLC cells, which induced apoptosis, only when IGF-1R signaling was blocked. Treatment with EGFR TKIs, but not with the EGFR antibody, induced EGFR:IGF-1R heterodimerization on cell surface and activation of the IGF-1R, resulting in the stimulation of PI3K/Akt/ mTOR pathway, promoting the de novo protein biosynthesis of survivin and EGFR, resulting in the survival of NSCLC cells. When IGF-IR and mTOR were blocked, treatment of EGFR-TKIs induced apoptosis in NSCLC cells, resulting in suppression of the NSCLC tumor growth. When we tested the expression of IGF-R and EGFR in human lung tissue, 9/14 tumor samples exhibited increased expression of EGFR and 7/9 samples showed a correlative increases in IGF-IR protein levels compared to their paired normal counterparts. Conclusions: These findings suggest that simultaneous targeting of EGFR and IGF-1R signaling pathways might be an effective therapeutic strategy against NSCLC.

No significant financial relationships to disclose.
In vitro chemosensitivity against enzastaurin correlates with gene expression of IL8 and GSK3-beta
Authors: Oberschmidt O. , Eismann U. , Lahn M. M. , Fleeth J. , Lüdtke F. , Struck S. , Schulz L. , Blatter J. , Ma D. , Hanauske A.
Published: March 2020

13046

Background: Enzastaurin (E) is an active antitumoral agent which selectively inhibits the β-isoform of protein kinase C (PKC-β). The compound blocks the enzyme’s ATP-binding site and signal transmission is abrogated resulting in the inhibition of neovascularization. The aim of the present study was to correlate gene expression with in vitro chemosensitivity of freshly explanted human tumor specimens. Such correlations in tumors taken directly from patients will help to rationally design subsequent clinical trials. Methods: Soft-agar colony forming assays were performed on freshly biopsied tumor cells exposed to various concentrations of E. Corresponding pieces of tumor specimens were shock-frozen and prepared for RNA isolation and cDNA generation followed by multiplex real-time PCR experiments. Gene expression data were correlated against cloning assay results. Results: Gene expression data of PKC-β1, PKC-β2, IL8RA, IL8RB, IL8, GSK3-β, and TGF-β were correlated against in vitro chemosensitivity pattern of E from 66 samples. After 1h-drug exposure gene expressions in sensitive versus resistant specimens were statistically significant with p = 0.013 for IL8 [median copy number (mcn): 1881 vs. 694; n = 66] and p = 0.012 for GSK3-beta (mcn: 1.6 vs. 7.0; n = 66). No correlation was detected for PKC-β1, PKC-β2, IL8RA, and IL8RB. Detection of TGF-β failed in most samples. Conclusions: Low expression of GSK3-β and high expression of IL8 correlate statistically significantly with increased in vitro sensitivity to E in freshly explanted human tumors. These findings may help direct further clinical development of this compound.

No significant financial relationships to disclose.
A phase Ib trial of ARQ 501, a selective checkpoint activator, in combination with docetaxel in patients with advanced solid tumors
Authors: Li C. , Nemunaitis J. , Senzer N. , Edelman G. , Glasner S. , Dombal G. , Dovholuk A. , Cunningham C.
Published: March 2020

13053

Background: ARQ 501 is a DNA damage checkpoint activator. It directly activates DNA damage response pathways without inducing primary DNA damage, resulting in selective apoptosis in a broad range of cancer cells. The cell death pathway of ARQ 501 is mediated by E2F1, and is independent of p53. In preclinical models, ARQ 501 demonstrates potent anticancer activity with a high therapeutic index. In these models, the addition of a variety of cytotoxic agents, including docetaxel, greatly enhances the anti-tumor activity of ARQ 501. Therefore, clinical investigation of this combination is warranted. Methods: Phase 1b dose escalation study in patients with advanced solid tumors to determine the maximum tolerated dose (MTD), pharmacokinetics, and preliminary antitumor activity of ARQ 501 in combination with docetaxel. Two schedules have been investigated. In one, ARQ 501 was administered for 5 consecutive days with a single docetaxel infusion at 50 to 100 mg/m2 given on day 3. In the other, a single dose of ARQ 501 was given in combination with a single docetaxel infusion on day 1. Both schedules were repeated every 3 weeks. Pharmacokinetic data were collected during study weeks 1 and 4 and tumor evaluations performed every 6 weeks. Results: To date, the first ARQ 501 treatment schedule has been given to 9 evaluable patients, and the second to 7 patients with a wide variety of tumor types. The combination is well tolerated and exhibits favorable, apparently linear, pharmacokinetics. ARQ 501 does not alter docetaxel pharmacokinetic parameters in patients. The most common grade 3 and 4 adverse events are neutropenia (4.9%) and anemia (4.4%). Of the 16 patients receiving at least 8 weeks of treatment, 12 have achieved a response of stable disease or better, including four responses (2 unconfirmed PRs based on RECIST, and 2 responses based on CA-125 decreases). Conclusions: The combination of ARQ 501 with docetaxel demonstrates clinical tolerability, favorable pharmacokinetics, and signs of antitumor activity. Further dose escalation is ongoing to determine a recommended phase II regimen.

[Table: see text]
Population pharmacokinetics (PK) of the vascular-disrupting agent 5,6-dimethylxanthenone-4-acetic acid (DMXAA) in cancer patients
Authors: Li J. , Jameson M. B. , Baguley B. C. , Pili R. , Baker S. D.
Published: March 2020

13076

Background: DMXAA (AS1404) is a small-molecule vascular-disrupting agent that selectively disrupts established tumor blood vessels. It is being evaluated in phase II trials in combination with cytotoxics. DMXAA is extensively metabolized, predominantly by UDP-glucuronosyltransferase (UGT) 2B7 and 1A9, and exhibits concentration-dependent protein binding. The study aim was to develop a population PK model for DMXAA and to examine covariates influencing drug disposition. Methods: 3,050 DMXAA total plasma concentrations were measured in 124 cancer patients receiving DMXAA monotherapy as a 20-min iv infusion weekly or every three weeks at doses from 6 to 4900 mg/m2 in 3 phase I trials. PK models were fitted to the data from all patients simultaneously using the program NONMEM. Potential covariates including age, body weight, height, body surface area (BSA), sex, and liver and kidney function tests were screened with general additive model analysis and tested in the PK model. Results: DMXAA concentration-time profiles after multiple-dose administration were well described by a 3-compartment model with saturable elimination (Michaelis-Menten kinetic) from the central compartment. BSA and sex were significant covariates on the volume of distribution of the central compartment (V1) and the maximum elimination capacity (Vm), respectively, accounting for 10 and 12% of interindividual variation (IIV) in these parameters. Population estimates for Vm, Km (concentration at which half-maximal elimination rate is achieved), and V1 were 122 × (1 + 0.502 × (2-SEX)) (μM/h) [females had lower Vm than males; SEX = 1 for males, 2 for females], 103 μM, and 8.15 × (BSA/1.8)0.69 (L), respectively. In the final model, IIV in Vm was 52%. Conclusions: DMXAA plasma disposition is characterized by a saturable elimination process. BSA-guided dosing for DMXAA is important. Evaluation of other covariates that reflect drug glucuronidation (e.g., UGT polymorphisms) may account for a greater fraction of the substantial unexplained PK variability. The population PK model will aid development of limited PK sampling strategies and investigation of PK-pharmacodynamic relationships in further trials.

[Table: see text]
Enzastaurin safety review: Data from phase I and phase II trials
Authors: Watkins V. , Hong S. , Lin B.
Published: March 2020

13077

Background: Enzastaurin, an oral inhibitor of PKC and PI3K/AKT pathways, blocks tumor angiogenesis, tumor cell proliferation and induces tumor cell death. Enzastaurin has demonstrated promise in phase I and II trials in patients (pts) with various advanced cancers. We report preliminary analysis of safety data of enzastaurin, obtained in previous trials, with particular consideration to long-term therapy. Methods: Data available to date from 3 different non-randomized, single-arm trials in pts with advanced cancers, diffuse large B cell lymphoma and mantle cell lymphoma, were analyzed with regards to treatment emergent adverse events (TEAEs). Severities were defined by NCI guidelines and investigators assessed the relatedness of AEs to enzastaurin. Results: A total of 135 pts (58 F; 77 M), mean age 64 years (range: 24–87) received 20–700 mg oral enzastaurin daily (103 pts at 525 mg). 36 pts were dosed for 0–30 days, 45 for 31–60 days, 35 for 61–180 days and 19 for ≥ 180 days. No deaths were related to enzastaurin. Chromaturia was the most common drug-related event (14%). No drug-related bone marrow suppression, Gr 3 neutropenia, thrombocytopenia or anemia was reported. TEAEs in ≥ 15% pts, or drug-related events ≥ Gr 3 reported in > 1 pt, are listed in the table . Other Gr 3 AEs, including edema, migraine and peripheral motor neuropathy were observed in 1 pt, each. Pts on therapy for ≤ 30 D reported highest number of AEs, consistent with the severity of their disease. Pts with exposure ≥ 180 days did not experience any drug related AE ≥ Gr 3. Conclusions: Enzastaurin is well tolerated across all doses and for extended durations with minimal AEs. In the 19 patients who received oral enzastaurin for ≥ 180 days, there was no indication of increased severity of any event. Given the severity of the disease and the absence of controlled studies for comparison, no event other than chromaturia or fecal discoloration (due to reddish-orange color of the active ingredient) appears to be definitively attributable to enzastaurin by this analysis.

[Table: see text]

[Table: see text]
Synergistic and selective inhibition of NSCLC cell growth via a caspase-independent cell death pathway by tumor suppressor 101F6 nanoparticles plus vitamin C in vitro and in vivo
Authors: Ohtani S. , Ueda K. , Jayanchandran G. , Xu K. , Minna J. D. , Roth J. A. , Ji L.
Published: March 2020

13086

Background: 101F6 is a candidate tumor suppressor gene on chromosome 3p21.3, a site of allele loss and genomic alterations were frequently found in many human cancers. We previously showed that enforced expression of wt-101F6 by adenoviral virus significantly inhibited tumor cell growth in 3p21.3-deficient NSCLC cells in vitro and in vivo. How 101F6 exerts this effect is largely unknown. Using a computer-aided structural and functional modeling, we recently identified 101F6 as a member of cytochrome b-561 protein family, which is involved in the regeneration of vitamin C. We hypothesized that under normal physiologic conditions, 101F6 protects cells from oxidative damage by regenerating antioxidant vitamin C and that in 101F6-deficient tumor cells, exogenous 101F6 facilitates vitamin C-mediated cytotoxic H2O2 formation. Methods and Results: We examined endogenous 101F6 expression in human NSCLC cell lines and tissue samples. All normal lung bronchial epithelial cells and fibroblasts but few lung cancers expressed 101F6. We investigated the combined effect of 101F6 and vitamin C on the cell growth: a nanoparticle-mediated wt-101F6 gene transfer plus a sub-pharmacologic concentration of vitamin C synergistically inhibited 3p21.3-deficient NSCLC cell growth but did not affect normal cell growth. We also used a human NSCLC H322 orthotopic lung tumor xenograft mouse model to evaluate the therapeutic efficacy of systemic injection of 101F6 nanoparticles and intraperitoneal injection of vitamin C. The growth of lung tumors was synergistically inhibited by the combination treatment (p<0.001). Furthermore, exogenous 101F6 promoted intracellular vitamin C uptake, leading to the vitamin C-mediated accumulation of H2O2 in the tumor cells, and these two agents synergistically killed the cells through caspase-independent apoptosis and autophagy cell death pathways. Conclusions: The synergistic and selective antitumor effect of 101F6 nanoparticles plus vitamin C may offer a useful tool for lung cancer prevention and intervention. This abstract is supported by grants from NCI (SPORE P50CA70907) and DOD (TARGET, DAMD17002–1-0706).

No significant financial relationships to disclose.
Marked synergism of cytotoxic agents with either a prenylation inhibitor or M-TOR inhibitor in acute leukemia cell lines: Potential clinical implications
Authors: Budman D. R. , Calabro A.
Published: March 2020

13103

Background: The most combinations of anticancer drugs are based upon empiricism. The potential permutations of drugs overwhelm the clinical trials system. Acute leukemia is sensitive to a variety of agents but relapses are common. Targeted agents are attractive new venues of therapy both as single agents and in combination with older agents. Isobologram median effect analysis allows up to three agents to be studied together in vitro to identify interesting combinations. We evaluated a commercially available statin, fluvastatin, to block prenylation which affects a variety of pathways, rapamycin and its experimental analogue RAD001 as M-TOR inhibitors to block downstream of the AKT pathway, and cytotoxic agents. Methods: The human leukemia cell lines AML-193 and KG-1 were obtained from ATTC (Rockville, MD), fluvastatin and RAD001 from Novartis Pharma, and the other agents from Sigma-Aldrich (St. Louis, MO). The IC50 of the single agent was determined by a 72 hr incubation of log growth cells using a MTT assay and the EZ-ED50 program (Perrella Scientific, Conyers, CA). The dosages of all agents were at clinically achievable concentrations. All reported values were the means of at least 3 experiments with each study using 4 wells per point. For isobologram analysis, a minimum of 8 concentrations of drug mixtures were studied above and below the IC50. Median effect CI values less than 1 are synergistic. Results: Doublets of fluvastatin with Ara-C (0.7), daunomycin (0.4), idarubicin (0.7), RAD001 (0.5), or rapamycin (0.3) demonstrated synergy. Doublets of RAD001 with Ara-C (0.3), daunomycin (0.7), or idarubicin (0.5) demonstrated synergy. Triplets of RAD001/daunorubicin/Ara-C, RAD001/daunomycin/fluvastatin, and RAD001/Ara-C/idarubicin all demonstrated marked synergy in both cell lines. Conclusion: A new potential non classical combination for further investigation is RAD001 or rapamycin with an inhibitor of prenylation such as fluvastatin. Additional potential combinations include cytotoxics with either fluvastatin or RAD001, and triplet combinations.

No significant financial relationships to disclose.
Cetuximab reversal of resistance to chronomodulated chemotherapy in heavily-pretreated patients with metastatic colorectal cancer (MCC) without amplification of epidermal growth factor receptor (EGFR) gene
Authors: Lévi F. A. , Karaboué A. , Bralet M. , Innominato P. , Herelle M. O. , Giacchetti S. , Adam R. , Jasmin C. , Guettier C. , Bouchahda M.
Published: March 2020

13104

Background Skin toxicity and EGFR amplification reportedly predicted for Cetux-reversal of MCC resistance to standard chemotherapy. We explored these relations for Cetux reversal of resistance to chronomodulated chemotherapy (Chrono) with irinotecan (CPT11, peak at 5 am), 5-fluorouracil-leucovorin (FU-LV, peak at 4 am) and oxaliplatin (l-OHP, peak at 4 pm). Methods: 40 pts with progressive MCC on 3 prior chemotherapy lines received weekly Cetux (250 mg/m2) and q2–3 wks CPT11-based (30 pts) or oxaliplatin-based (10 pts) Chrono. Toxicities and response were assessed q2–3 wks and q2 months (mo) respectively. Tumor EGFR expression was determined by immunohistochemistry for all pts and gene copies by fluorescent in situ hybridization (FISH) for 25 pts. Results: All pts had prior progression on CPT11, FU-LV and l-OHP. Median age: 61 yo; M/F: 24/16; WHO PS 0/1/2: 27/9/4; colon/rectum: 21/19; M sites 1/2/≥ 3: 12/15/13; liver/lung: 32/29; pre-existing peripheral sensory neuropathy grade ≥ 2: 11 pts. A median of 8 courses of Cetux-Chrono was given (1 to 22). Three pts withdrew for grade 4 allergy. 37 pts were assessable for other toxicities and 36 for response (1 too early). Gr 2–3 acneiform skin rash occurred in 27 pts (73%), with gr3 in 11 pts (29.7%). Other main gr 3–4 toxicities were diarrhea (27% of the pts), neutropenia (24.3%) and neuropathy (21.6%). Disease progressed in 12 pts (33.3%) and was controlled in 24 pts (66.7%) including 13 stable disease (36.1%) and 10 objective responses (8 PR and 2 CR) - 27.8% [95% C.L. 12.9 to 42.7]. EGFR was expressed in 32 pts (80%), with >10% + tumor cells for 14 pts (35%). 3/6 EGFR- pts (50%) and 7/32 EGFR+ pts (21.9%) responded. No gene amplification was documented in the 25 tumors (7 responders). A positive correlation between skin rash and disease control was suggested (p from χ2 = 0.08). Median progression free and overall survival (mo) are 5.3 [2.9–8.4] and 11.1 [8.6–13.6] respectively. Conclusions: Cetux partly alleviated MCC resistance to Chrono, independently of tumor EGFR protein expression or gene amplification, possibly through interfering with TGFα. This EGFR ligand carries poor prognosis for MCC and alters circadian clock function.

[Table: see text]
Thymidylate synthase and MDR1 mRNA levels predict response to chemotherapy with S-1 in metastatic colorectal cancer
Authors: Fujita I. , Hayashi K. , Uchida K. , Kuramochi H. , Kudo K. , Miyakura S. , Yamamoto M. , Takasaki K.
Published: March 2020

13107

Background: To test the hypotheses of whether the relative mRNA expression of the thymidylate synthase (TS) gene and multidrug resistance 1 (MDR1) gene are associated with response to chemotherapy with S-1 in metastatic colorectal cancer. Methods: Eighteen patients with progressive stage IV CRC were treated with S-1 twice daily (BSA = 1.5 m2, 60 mg/day) for 28 days, followed by a 2-week period rest. cDNA was derived from frozen tumor specimens to determine TS and MDR1 mRNA expression relative to the internal reference gene GAPDH using fluorescence-based, real-time reverse transcriptase polymerase chain reaction (Taqman) system. Results: The median TS gene expression level from 18 CRC tumors was 0.94 × 10(−3) (minimum expression, 0.15 × 10(−3); maximum expression, 8.0 × 10(−3)), and the median MDR1 gene expression level was 7.75 × 10(−3) (minimum, 0.83; maximum, 41.5 × 10(−3)). The gene expression cutoff values for chemotherapy nonresponse were 1.0 x 10(-3) for TS and 10.0 × 10(−3) for MDR1. The response rate for patients with TS < or = 1.0 × 10(−3) (10 of 18 patients) was 60% (6/10), compared with 0% (0/8) for patients with TS greater than 1.0 × 10(−3) (P = 0.013). Patients with MDR1 expression < or = 10.0 × 10(−3) (13 of 18 patients) had a response rate of 46% (6/13), compared with 0% (0/5) for patients with MDR1 expression greater than 10.0 × 10(−3) (P = 0.11). Regarding the combination of TS and MDR1 Expression, low TS and MDR1 expression levels were detected in 8 (44%) of the patients, and 10 patients (56%) had a high TS and/or MDR1 expression level. The response rate was 75% (6/8) for the low TS and MDR1 expressors and 0% (0/10) for the high TS and/or MDR1 expressors (P = 0.001). Conclusions: These data suggest that intratumoral TS and MDR1 mRNA expression levels are independent predictive markers of response to S-1 chemotherapy in metastatic colorectal cancer.

No significant financial relationships to disclose.
Epidermal growth factor receptor (EGFR) mutation rate in advanced ovarian cancer (AOC): Results of a prospective study in Caucasian patients (pts)
Authors: Mustea A. , Koensgen D. , Zeillinger R. , Castillo-Tong D. C. , Sun P. , Mobus V. , Blohmer J. , Pirvulescu C. , Lichtenegger W. , Sehouli J.
Published: March 2020

13127

Background: The EGFR is over expressed in 55% to 98% of advanced epithelial ovarian carcinoma. Different studies demonstrated EGFR status as an independent prognostic factor for OC. Recent studies in non small cell lung cancer suggest that the presence of the clinical response to tyrosine kinase inhibitors (e.g. ZD 1839) correlates with the somatic mutations in the kinase domain of EGFR, exons 18–21. For pts with OC data are not available on EGFR gene mutation. Methods: Shock-frozen samples from 32 patients (pts) with primary of ovarian cancer were stratified in two groups according to disease-free interval: ≤6 months (17 pts.) and <6 months (15 pts.). All pts were prospectively collected within Tumor bank Ovarian Cancer Project. Patient collective consisted only from west European Caucasian women. Additionally, 9 commercial available ovarian cancer cell lines (TOV-90, TOV-112D, TOV-21G, OVCAR-3, A2780, A2780 ADR, ES-2, SK-OV-3, and Caov-3) and 32 established ovarian cancer lines were analysed. Exon sequencing of genomic DNA was used to detect L858R deletion mutations of EGFR within exons 21 of the kinase domain. PCR and capillary electrophoresis (Chip-Format) were used to analyse 15 bp deletion in Exon 19. We focused on descriptive analysis. The Log-Rank test was applied to confirm statistically significance (p-value of <0.05). Results: Overall, 74.6% of the pts. were diagnosed FIGO stage III-IV. Median follow-up period was: 14.17 month (range: 2–42 months). Whether in cell lines, nor in tumor samples, stratified to response of platinum therapy any mutation of EGFR gene was observed. Conclusions: Our study indicates that the prevalence of mutation in the kinase domain of EGFR, exons 19 and 21 seems to be very low in pts. with AOC. Further studies should investigate other ethical groups of pts.

No significant financial relationships to disclose.
Intervertebral administration of 153Samarium with kyphoplasty for vertebral metastases
Authors: Ashamalla H. , Cardoso E. , Weng L. , Rafla S. , Macedon M. , Panigrahi N.
Published: March 2020

13136

Background: Kyphoplasty is an effective procedure to alleviate pain in metastatic vertebrae. 153Samarium-EDTMP (153Sm) is used for palliative treatment of multiple bone metastases. The hematological toxicity is the main limiting factor. Objectives: Study the feasibility of intervertebral administration of 153 SM with Kyphoplasty. Assess procedure related morbidities specifically haematological since a direct vertebral route is planned though to a much smaller dose. Methods: IRB approval for an off label use of 153Sm-EDTMP was obtained. Six patients with documented vertebral bone metastasis and pathological compression fractures were studied (Thoracic in 2 and lumbar in 4). Primary cancer was lung in 2, prostate (1), maxillary sinus (1), myeloma (1) and colon (1). Serial dilution of 153 SM was performed to obtain 2 mci/1ml. Kyphoplasty procedure was carried out using the known protocol. 2 mCi of 153Sm was admixed with the bone cement and administered under tight radiation safety measures. Serial nuclear body scans were obtained. Pain assessment was evaluated using analog pain score at 1, 7 and 30 days. Serial blood counts are followed. Results: All patients tolerated procedure well. No immediate procedure(s) related morbidities were noted. One case was not technically satisfactory and no significant uptake was detected in the injected vertebra. Nuclear scan done on 0 revealed excellent radiotracer uptake in the other 5 vertebrae injected. The absorbed radiotracer was seen to target other skeletal metastatic lesions in 2 patients. There was no significant change in WBC, HCT or Platelets seen in blood counts obtained over one month post-procedure. Except for the first patient, no appreciable radiation leakage was encountered. Pain score was reduced from a mean of 8.5 to 3.2 during the first week and was maintained below score of 4 at one month assessment. Conclusions: The combination of intervertebral administration of 153SM and Kyphoplasty seems to be well tolerated. No haematological side effects were encountered; probably due to the use of only 2 mci compared to over 70 mci in intravenous administration. Pain control seems to be satisfactory at areas treated. Analysis of procedure related radiological changes is underway.

No significant financial relationships to disclose.
Enzastaurin a protein kinase Cbeta-selective inhibitor, inhibits the growth of SCLC and NSCLC cell lines
Authors: Nakajima E. , Helfrich B. , Chan D. , Zhang Z. , Hirsch F. R. , Chen V. , Ma D. , Bunn P. A.
Published: March 2020

13138

Background: PKCβ is a member of the PKC family of serine-threonine protein kinases involved in tumor cell proliferation and apoptosis. Inhibition of PKCs induced differentiation and enhanced chemotherapy. PKCβ activation is required for tumor-induced angiogenesis. The PKCβ-selective inhibitor enzastaurin, originally developed as an antiangiogenic agent, inhibited tumor cell proliferation in prostate, colon and glioblastoma cell lines in vitro. In NSCLC lines, enhanced phosphorylation and altered PKC expression was demonstrated. In SCLC lines specific PKC isoforms were associated with cisplatin resistance. Methods: The growth inhibitory effects of enzastaruin were evaluated by 6-day MTT assays; the cell cycle effects by FACS analysis; the effects on downstream phophorylated signaling molecules by western blotting. Results: Enzastaurin inhibited the growth of 11 SCLC lines (IC50s 3–10 μM) and 4 NSCLC cell lines (IC50s 3–10 μM). An increase of 7–31% of cells in the G2/M phase of the cell cycle compared to untreated control was observed following 48 hour exposure to the IC50 dose of enzastaurin in both SCLC and NSCLC cell lines. PKCβ has been shown to phosphorylate both GSK3β and Akt. A 24-hour IC50 enzastaurin exposure significantly reduced phosphorylation of GSK3β (Ser9) in both SCLC and NSCLC lines. No changes were observed in phospho-AKT (Thr308) in either SCLC or NSCLC cell lines. Phospho-ribosomal protein S6 (Ser240/244) was also reduced in both SCLC and NSCLC cell lines. Potential synergy was studied between enzastaurin and pemetrexed in SCLC and NSCLC lines and the results were analyzed using the Calcusyn Program by Chou and Talalay. Synergistic (CI <1) to additive interactions were observed between pemetrexed (IC20–70) and enzastaurin (≤ IC50) in both SCLC lines (N = 3) and NSCLC lines (N = 2). Conclusions: We conclude that enzastaruin produces in vitro growth inhibition of SCLC and NSCLC cell lines through inhibition of GSK3β ser9 phosphorylation and has synergistic growth inhibition with pemetrexed.

[Table: see text]
OncoVEX: A family of oncolytic herpes simplex viruses optimised for therapeutic use
Authors: Coffin R. S. , Liu B. , Han Z. , Assenberg M. , Thomas S. , Hu J. , Simpson G.
Published: March 2020

13139

HSV in which ICP34.5 is deleted directs tumour selective cell lysis and has proven safe in Phase I clinical trials. To produce oncolytic HSV with enhanced anti-tumour properties, we have deleted ICP34.5 from a clinical isolate of HSV-1, which enhances the tumour cell killing capabilities of the virus, deleted ICP47 (which blocks antigen presentation), and inserted the gene encoding GM-CSF. This aimed to maximize anti-tumour immune responses following intra-tumoural injection and provide an in situ, patient-specific, anti-tumour vaccine, combined with oncolysis. In vivo, both injected and non-injected tumours could be cured and animals were then protected against tumour cell challenge. A Phase I clinical trial with the virus (OncoVEXGM-CSF) has been conducted including patients with cutaneous or sub-cutaneous deposits of a number of tumour types (Lead Investigator: Professor Charles Coombes, Hammersmith Hospital, London). This demonstrated the virus to have a good safety profile, the main side effects being ‘flu-like symptoms, similar to those which have previously been observed with other oncolytic products. Virus replication and GM-CSF expression was demonstrated together with inflammation, flattening and necrosis of injected lesions which was in some cases considerable and which was also in some cases observed in lesions which had not themselves been injected. In all cases where necrosis was observed in biopsies, this correlated with areas of staining for HSV, suggesting the virus had caused the effect. Following this promising data, Phase II studies are underway in multiple tumour types. In addition to OncoVEXGM-CSF, further versions of OncoVEX expressing other active genes have been constructed and tested in pre-clinical models. These include a virus expressing TNF∝, intended to be synergistic with radiotherapy, and versions of the virus expressing a pro-drug activating gene combined with the delivery of a fusogenic glycoprotein designed to maximize the properties of the virus for local tumour control. Each of these have shown promising results in pre-clinical tumour models, including in combination with chemotherapy and radiotherapy where benefits which are at least additive have been demonstrated.

[Table: see text]
Compassionate treatment (CTr) with re-irradiation, carboplatin (Cb) and Cetuximab (C) in very advanced head and neck cancer (HNC)
Authors: Numico G. , Colantonio I. , Heouaine A. , Occelli M. , Fea E. , Russi E. G. , Vigna Taglianti R. , Lanzillotta L. , Merlano M. C.
Published: March 2020

13153

Background: Extremely locally advanced HNC is a dramatic clinical situation, causing pain, repeated infections, speech and swallowing impairmen, malnutrition and cosmetic disasters. Re-irradiation may partially palliate some pt, but the low dose of radiation that can safely be delivered limits this approach. Recently the combination of C and radiation has shown improvement of clinical outcome over radiation alone. Methods: We started a CTr with re-irradiation concurrent with Cb and C in pts with heavily symptomatic, pretreated HNC. CTr consisted of: RT (30 Gy) 2 Gy/d, 5 d/week given every other week; Cb AUC 6 d 1, 22 and 43; C, 400 mg/m2 loading dose and 250 mg/m2 maintenance, weekly until day 43 (6 administrations). Results: From January 04 to June 05, 8 pts pre-treated with surgery (6 pts), radiation (8) and chemotherapy (6) entered the CTr. All pts but one had bulky soft tissue invasion and 2 had skin ulceration. One had only bulky nodal relapse. One pt died after the 2nd week due to pneumonia. The other pts had a fast response to CTr: within 5–7 d from the start of therapy they reached a clinical benefit (major or complete pain relief, improvement of swallowing and/or neck movements) and subsequently a rapid tumor shrinkment. All pts achieved an objective response (complete response in 2). No significant local or systemic toxicity occurred in any of the pts, but a mild skin rush. The first 4 pts did not receive further treatment after CTr. These pts showed tumor re-growth within 4–6 weeks from the end of treatment. However one of them underwent salvage surgery and is alive and disease free 14 months after CTr. The 3 other pts received maintenance treatment with weekly C until disease progression. One of them died due to cancer unrelated causes without PD 11 weeks after RT. The 2nd pt developed PD during RT. The 3rd pt is progression-free and alive at 30 weeks from the end of RT. Overall, TTP of the 7 pts ranges between 2 and 7+ months (median 3); survival from the end of RT ranges between 3 and 14+ months (median 7+). Conclusions: CTr with re-irradiation, Cb and C seems feasible even in this poor prognosis, very advanced HNC. No additional toxicity was recorded by adding C. Palliation was achieved in almost all pts and the results deserve further investigations.

No significant financial relationships to disclose.
Arsenic trioxide for treatment of myelodysplastic syndrome: An experience from India
Authors: Mukhopadhyay Sr. A. , Das S. , Dey S. , Choudhury R. , Ahmed R. , Gupta P. , Dhara A. , Chakraborty S. , Halder B. , Mukhopadhyay S.
Published: March 2020

13156

Background: Myelodysplastic Syndrome (MDS) is a heterogeneous group of clonal diseases of the haematopoietic stem cells. The hallmark of the disease is ineffective haematopoiesis characterized by dysplasia with incomplete maturation and progressive increase in the percentage of myeloblast. No standard treatment is currently available for MDS. The early clinical experience has confirmed the activity of arsenic trioxide in MDS. The drug is able to induce differentiation and apoptosis and to inhibit cell proliferation or angiogenesis. It has the potential to be active in tumour models in MDS. The preliminary result of ongoing studies conducted in patients with MDS suggests that arsenic trioxide produces haematological improvement including durable transfusion independence in 30% of patients. The aim of our study was to see the response of MDS with arsenic trioxide and to see the toxicity profile of arsenic trioxide in Asian Indian population. Methods: During period from July 2005 to December 2005 we selected consecutive 10 patients of MDS in Refractory Anaemia, Refractory anaemia with ringed sideroblasts, Refractory anaemia with blast excess, Refractory anaemia with blast excess in transformations and chronic myelo monocytic leukemia phases. All patients had performance status more than 60%, some karyotypic abnormalities & in cytopenic phase. Median age of the patients 65 years (range 42 to 70 years). All patients were treated with arsenic trioxide 10mg (Alkem/India) daily for 2 hours infusion 28 days. In 15 days interval 3 courses were repeated. Response assessments were done by haematological, cytogenetic & quality of life assessment. All patients were evaluated after 3 courses of arsenic trioxide. Result: Sixty percent (6 patients) patients had shown major haematological response, forty percent minor & twenty percent has major cytogenetic response. Twenty percent of the patients has disease progression where as 20% has stable disease. The only mild adverse effects were seen in forms of nausea, vomiting, diarrhea, abdominal pain & dermatitis in 30% of patients. Only one patient (10%) had QT prolongation in ECG. Conclusion: We concluded that arsenic trioxide is very useful drug in myelodysplastic syndrome. It is also well tolerated in Asian Indian Population.

No significant financial relationships to disclose.