bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5043: Population pharmacokinetic model for OT-101 - A TGF-β2-specific antisense oligonucleotide in cancer patients
Authors: Wang Wen , Ng Kevin , Nam David , Trieu Vuong , Hwang Larn
Published: July 2017

Background: OT-101 (Trabedersen) is a phosphorothioate antisense oligodeoxynucleotide specifically inhibiting the expression of transforming growth factor-beta 2 (TGF-β2), whose overexpression is a pivotal factor for malignant progression in solid tumors. In the clinical Phase I/II study, plasma pharmacokinetic (PK) profile of OT-101 administered intravenously was evaluated in patients with advanced tumors. A population PK model was built to further understand the factors contributing to the variability in PK of OT-101.

Methods: A total of 61 patients with pancreatic cancer (n=37), malignant melanoma (n=19), or colorectal carcinoma (n=5) were treated with OT-101 with escalating doses in 2 treatment schedules (1st schedule: 7-days on, 7-days off; 2nd schedule: 4-days on, 10-days off; up to 10 cycles). The PK analysis was only performed on Cycle 1 and, when applicable, Cycle 2. Blood samples were collected from patients at planned time points from before start of infusion to 7 days (1st schedule) or 10 days (2nd schedule) after stop of infusion. The plasma concentration data of OT-101 were used to build a population PK model using Phoenix NLME software. The influence of age, gender, body mass index (BMI), body weight (BW), height, cancer type, and treatment schedule as covariates on PK was evaluated.

Results: With exclusion of protocol deviations, a total of 1444 plasma sample concentration data from 100 patient cycles were examined. The concentration time course of OT-101 was best described by a two-compartment model. The model estimated the PK parameters as follows: total body clearance, 0.17 mL/h; distribution volume of the central compartment, 4.69 L; inter-compartmental clearance, 3.31 L/h; distribution volume of the peripheral compartment, 5046.44 L. BW was identified as a covariate on OT-101 inter-compartmental clearance, with KBW as -ve 1.48.

Conclusion: The PK parameters of OT-101 were best described by a two-compartment model. OT-101 was largely distributed in the peripheral tissues. The influence of age, gender, BMI, height, cancer type and treatment schedule on the PK of OT-101 was not identified. The model will be used for a sparse population PK study during the planned phase III trial of OT-101 in pancreatic cancer.

Citation Format: Wen Wang, Kevin Ng, David Nam, Vuong Trieu, Larn Hwang. Population pharmacokinetic model for OT-101 - A TGF-β2-specific antisense oligonucleotide in cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5043. doi:10.1158/1538-7445.AM2017-5043
Abstract 5046: Exposure-efficacy (OS) analysis of tremelimumab in unresectable malignant mesothelioma
Authors: Baverel Paul , Roskos Lorin , Tatipalli Manasa , Lee Nancy , Stockman Paul , Taboada Maria , Vicini Paolo , Horgan Kevin , Narwal Rajesh
Published: July 2017

Purpose: Tremelimumab is a fully human anti-CTLA-4 IgG2 monoclonal antibody that enhances human T-cell activation. Tremelimumab was evaluated in a Phase IIb (DETERMINE), randomised, double-blind, placebo-controlled study in patients with unresectable pleural or peritoneal malignant mesothelioma, randomised (2:1) to receive either tremelimumab (10 mg/kg, seven doses Q4W followed by Q12W) or placebo. The study demonstrated no clinically meaningful differences in overall survival (OS). The primary objectives of this analysis were to evaluate the relationship of exposure with OS, and the impact of potential confounders.

Methods: A population PK model was developed to estimate and derive PK exposure metrics (area under the curve at steady state [AUCss] or clearance [CL]) for exposure-OS analysis. Impact of potential confounders was evaluated using graphical and exploratory approaches. Factors including body weight, age, gender, race, ECOG status, anatomical site (pleural or peritoneal), line of therapy, EORTC status, tumour histology, baseline tumour size, LDH, and CRP were evaluated. The analyses were performed using NONMEM 7.2 and R software.

Results: The population PK included 376 patients and 1328 post-first dose PK concentrations. PK was consistent with previous knowledge and low incidence of anti-drug antibodies was observed. A 2-compartment linear PK model adequately described the data. Tremelimumab CL and volume of distribution (V1) were 310 mL/day and 3.85 L, with moderate variability of ~38% and ~32%, respectively. There was an apparent exposure-OS relation when stratified by AUCss. However, at least 3 factors (gender, CRP, and baseline tumour size) were statistically significant PK predictors (p<0.05 on CL) indicating multi-dimensional confounding effect. Higher baseline tumor size, higher CRP levels and males were associated with lower PK exposure of tremelimumab.

Conclusions: The observed apparent exposure-OS relationship is the result of imbalance in prognostic factors impacting OS rather than a true association of exposure with efficacy.

Citation Format: Paul Baverel, Lorin Roskos, Manasa Tatipalli, Nancy Lee, Paul Stockman, Maria Taboada, Paolo Vicini, Kevin Horgan, Rajesh Narwal. Exposure-efficacy (OS) analysis of tremelimumab in unresectable malignant mesothelioma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5046. doi:10.1158/1538-7445.AM2017-5046
Abstract 5055: Multiplatform metabolomics analysis of growth arrest in pancreatic tumor xenografts
Authors: Wainer Irving W. , Dudzik Danuta , Bernier Michel , Barbas Coral
Published: July 2017

R,S-4-Methoxy-1-naphthylfenoterol, MNF, inhibits the G protein-coupled receptor GPR55. In this study, we determine the effect of MNF on human PANC-1 pancreatic tumor growth in mice and apply multiplatform metabolomics analysis to identify pathways associated with growth arrest.

Methods: Female Balb/c nude mice, 6-8 weeks old, 18-20g, were inoculated subcutaneously with 5 x 106 PANC-1 cells. On Day 8, mice were placed in groups of 10 using random block design based upon tumor volume. Mice received daily ip injections of vehicle or 40mgxkg-1 MNF 5 days/week for 3 treatment cycles. Mice were monitored daily and tumor volumes measured at beginning and end of each dosing cycle. On Day 33, mice were euthanized, and plasma samples and tumors collected. Tumor tissue was homogenized, extracted and analyzed using liquid chromatography-QTOF-MS, capillary electrophoresis-TOF-MS and gas chromatography-EI-Q-MS. Differences between groups were evaluated by unpaired t-test or Mann−Whitney test with post hoc Benjamini-Hochberg correction. Statistical significance was set at P≤0.05. Compound identification was accomplished using online databases and in-house standards

Results Tumor volume increased in vehicle-treated mice by ~700%, 142 ± 8 mm3 to 957 ± 79 mm3 and only ~250% in MNF-treated mice, 143 ± 8 mm3 to 259 ± 27 mm3. On Day 33, MNF was not detected in plasma but accumulated in tumor tissues, 43.9 ± 32.7 ng/g. Plasma L-lactate levels were reduced from 3.29 ± 0.66 mmol/L το 2.81 ± 0.60 mmol/L, P≤0.001. Differences in tumor tissue metabolome were observed with MNF compared to vehicle, which was reflected by significant, P≤0.05, changes in relative metabolite signals. MNF treatment was accompanied by the disruption of pyrimidine nucleotide biosynthesis at uridine 5'-monophosphate, UMP, and increased UMP degradation. Moreover, the +237% increase in ophthalmic acid and +95% increase in its precursor 2-aminobutyrate indicate higher oxidative stress and the +51% increase in 2-hydroxyproline suggest greater HFI-1α proteolysis upon MNF treatment.

Conclusions These results are consistent with our previous observations in PANC-1 cells showing that GPR55 inhibition attenuates activation of the EGFR-MEK-ERK, Wnt-β-catenin and PI3K/AKT pathways resulting in lower cyclin D1 expression and halting the cell cycle in G1. The reduction in HIF-1α expression and glycolytic flux in MNF-treated PANC-1 cells is in line with the decline in L-lactate plasma levels in MNF-treated mice. The data from this study indicate that MNF may be useful in the treatment of pancreatic cancer.

Citation Format: Irving W. Wainer, Danuta Dudzik, Michel Bernier, Coral Barbas. Multiplatform metabolomics analysis of growth arrest in pancreatic tumor xenografts [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5055. doi:10.1158/1538-7445.AM2017-5055
Abstract 5062: Development of a first in class inhibitor of BET bromodomains and dopamine receptor 2
Authors: Yoshioka Makoto , Chauhan Jay , Fletcher Steven , Strovel Jeffrey W.
Published: July 2017

ConverGene has developed a first-in-class dual-active small molecule inhibitor that i) inhibits BET family of bromodomain-containing proteins, and ii) antagonizes dopamine receptor D2 (DRD2). BET protein family includes BRD4, an epigenetic reader protein that mediates expression of MYC oncogene. Thus, BRD4 is considered as a cancer therapeutic target to indirectly suppress MYC expression. In addition to being a therapeutic target for psychiatric diseases, DRD2 is emerging as a potential therapeutic target in neuroendocrine tumors, subsets of pancreatic ductal adenocarcinoma and small cell lung cancer. Our lead compound showed high activity in a binding test against BRD4 (Ki = 34 nM); exhibited high bioavailability upon oral administration; profoundly suppressed MYC expression both in vitro and in vivo; inhibited growth of AML and solid tumor cells in xenograft models; potently inhibited both isoforms of DRD2 (IC50 0.1 µM); and interfered with DRD2/β-arrestin/Akt pathway in vitro. Therefore, our BRD4/DRD2 dual-active compounds may hold promise as a novel class of therapeutics that interferes with both cancer growth and maintenance by simultaneously interfering with MYC and DRD2 pathways. We currently are investigating these dual-active compounds in multiple in vitro and in vivo models and expect to report the outcomes at the AACR Annual Meeting in 2017.

Citation Format: Makoto Yoshioka, Jay Chauhan, Steven Fletcher, Jeffrey W. Strovel. Development of a first in class inhibitor of BET bromodomains and dopamine receptor 2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5062. doi:10.1158/1538-7445.AM2017-5062
Abstract 5066: Combination treatment with SAHA and 5-Azacytidine (Decitabine) induces apoptosis and suppresses tumor growth in preclinical models of chondrosarcoma
Authors: Sheikh Tahir , Patwardhan Parag , Schwartz Gary K.
Published: July 2017

Chondrosarcomas are the second most frequently occurring type of bone malignancy, and account for approximately 25% of all bone sarcomas. They are often highly aggressive neoplasms that rapidly progress and eventually recur and give distant metastases. They are largely considered to be resistant to conventional chemotherapy and radiotherapy. Several studies have reported that targeting epigenetic mechanisms including DNA methylation or histone acetylation are novel approaches for the treatment of some human cancers. Previous reports have shown that the interaction of DNA methylation and histone modification regulates gene expression. In the present study, we hypothesized that concurrent inhibition of histone acetylation and DNA methylation could result in decreased viability of chondrosarcoma cells both in vitro and in vivo. To test this, we used a panel of chondrosarcoma cell lines including IDH wild type (CH2879), IDH1 mutant (JJ012) and IDH2 mutant (CS1) cell line. Results from our in vitro proliferation assay showed that combination of sub-IC50 concentrations of the DNA methyltransferase (DNMT) inhibitor Decitabine (5-AZA-dC) and histone deacetylase (HDAC) inhibitor (SAHA) resulted in decreased cell viability of all the three chondrosarcoma cell lines tested when compared to either drug alone. Western blot analysis showed induction of cleaved Poly-ADP Ribose Polymerase (PARP), a known marker of apoptosis. Consistent with augmented DNA damage, combination of Decitabine and SAHA markedly increased the levels of phospho-H2AX, a DNA damage marker, and pro-apoptotic BH3-only proteins such as Bim. Combination treatment also resulted in increased induction of histone acetylation (AcH3) and increased expression of E-Cadherin. Previous reports have shown that E-cadherin is critical for apoptosis and depletion and inhibition of E-Cadherin impairs apoptosis induction via DR4 and DR5 death receptors. To this end, we showed that induction of cleaved PARP by Apo2L/TRAIL stimulation decreased over time when E-Cadherin was knocked down by siRNA. Xenograft studies using IDH2 mutant (CS1) tumor model showed a significant suppression of tumor volume when animals were treated in combination with SAHA and Decitabine compared to single agent treatments. Taken together, our data strongly suggests that combination treatment with SAHA and Decitabine is a novel treatment approach and merits evaluation in the treatment of chondrosarcoma.

Citation Format: Tahir Sheikh, Parag Patwardhan, Gary K. Schwartz. Combination treatment with SAHA and 5-Azacytidine (Decitabine) induces apoptosis and suppresses tumor growth in preclinical models of chondrosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5066. doi:10.1158/1538-7445.AM2017-5066
Abstract 5068: A novel dual action inhibitor of histone deacetylase and demethylase in melanoma
Authors: WU Muzhou , Kalin Jay , Ryu Byungwoo , Cole Philip , Alani Rhoda
Published: July 2017

Epigenetic agents have drawn great attention as anti-cancer therapies, with several HDAC inhibitors approved for a subset of hematologic malignancies. One of the biggest challenges in targeting epigenetic mechanisms of tumorigenesis is the wide spectrum of effects which restrict the therapeutic window for these compounds. We have developed a series of potent small molecule inhibitors with specificity towards the CoREST epigenetic corepressor complex through a dual-action mechanism targeting LSD1 and HDAC1. These compounds show a unique profile of pharmacologic action with an improved therapeutic window in a variety of cell types. Screening of tumor cell lines for growth inhibitory effects revealed variable efficacy in a broad spectrum of cancers with the most consistent and potent effects seen in human melanomas. The growth of a number of melanoma cell lines was found to be potently inhibited by one of these compounds, Corin 2; however, primary human melanocytes were relatively resistant to this agent. Transcriptomic analysis revealed that Corin2 was a more potent inducer of tumor suppressor genes compared to the parent HDAC and LSD1 compounds. Genetic knockdown of CoREST or LSD1 in cancer cell lines abolished the differences in potency of Corin2 vs. the parent HDAC inhibitor, Entinostat, suggesting that Corin2's favorable pharmacologic effects rely on an intact CoREST complex. Corin2 was also effective in slowing tumor growth in a melanoma mouse xenograft model. These dual action inhibitors demonstrate a novel, potent, and specific therapeutic approach to targeting epigenetic pathways in human melanomas which may lead to improved therapeutic benefits in patients with advanced disease.

Note: This abstract was not presented at the meeting.

Citation Format: Muzhou WU, Jay Kalin, Byungwoo Ryu, Philip Cole, Rhoda Alani. A novel dual action inhibitor of histone deacetylase and demethylase in melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5068. doi:10.1158/1538-7445.AM2017-5068
Abstract 5073: Determination of sensitivity to BET and HDAC inhibitors in lung cancer cell lines
Authors: Verdura Sara , Torres-Diz Manuel , Sanchez-Cespedes Montserrat
Published: July 2017

Purpose: To determine the sensitivity of lung cancer cell lines to different combinations of BET and HDAC inhibitors and to inquire possible associations with the genetic and molecular profiles.

Methods: A panel of lung cancer cell lines were treated in vitro with BET and HDAC inhibitors. The capability of each compound, administered individually or in combination (IC50), to inhibit cell proliferation was assessed by MTT assays. Drug interaction analysis was performed using Compusyn software. The protein and gene expression levels were determined using western blot and real time PCR, respectively.

Results: We found that about 70% of the cell lines tested were sensitive to BET inhibitors and about 60% responded to HDAC inhibitors. In both cases, increased sensitivity was accompanied by changes in cell morphology, a reduction of the MYC protein levels and an increase in the levels of apoptotic-related markers. We also determined the effects of the combination of these epigenetic treatments in cell growth. Our results showed a synergic effect in about 75% of the cell lines which was also correlated with dramatic changes in cell morphology, in apoptosis and in decreasing the levels of the MYC protein. Finally, we determined the possible correlation of the sensitivity to these epigenetic drugs and the expression levels of some genes implicated in lung carcinogenesis.

Conclusions: Our data showed that lung cancer cells responded to BET and HDAC inhibitors and that the combination of both drugs has a synergistic effect in cell growth inhibition, suggesting that this could be an effective treatment in lung cancer. Our data also point out to certain genes as potential biomarkers to determine sensitivity to these compounds.

Citation Format: Sara Verdura, Manuel Torres-Diz, Montserrat Sanchez-Cespedes. Determination of sensitivity to BET and HDAC inhibitors in lung cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5073. doi:10.1158/1538-7445.AM2017-5073
Abstract 5075: Phenotypic human tumor cell line profiling and predictive biomarker analysis for epigenetic inhibitors
Authors: Warrior Usha , Parry Jesse J. , Burkart Samantha C. , Cavedine Lee R. , Cracchiolo Alyssa M. , Epkins Brogan A. , Garner Steven M. , Sindelar Timothy J. , Wageman Charles R. , King Alastair J.
Published: July 2017

In recent years, epigenetic target inhibitors have emerged as viable therapeutics for treating various types of cancers. Testing of the epigenetic inhibitors in cell proliferation assays revealed that inhibition of many epigenetic targets resulted in a significantly delayed drug response that is not revealed by conventional, 72-hour cell growth assays. OncoPanel LT™ from Eurofins Pharma Discovery Services is developed specifically for evaluation of compounds with a protracted mechanism of action, and consists of a large collection of cell lines with broad cancer type and subtype representation, and extensive genomic characterization. We have tested decitabine, a DNA methyltransferase inhibitor, panobinostat, a histone deacteylase inhibitor, GSK-343, an EZH2 inhibitor, GSK-J4, a JMJD3/UTX inhibitor, and JQ1, a BRD2/3/4 inhibitor in OncoPanel LT™. These epigenetic inhibitors were incubated for 10 days with more than 200 human tumor cell lines and the inhibition data were analyzed to determine IC50, EC50, and GI50 values. These inhibition data, as well as the genomic features of the cell lines, were utilized to perform univariate genomic analysis to identify predictive biomarkers of response. We will discuss the activity of compounds against this cell line panel, as well as statistically significant biomarkers of response, based on the univariate genomic analysis.

Citation Format: Usha Warrior, Jesse J. Parry, Samantha C. Burkart, Lee R. Cavedine, Alyssa M. Cracchiolo, Brogan A. Epkins, Steven M. Garner, Timothy J. Sindelar, Charles R. Wageman, Alastair J. King. Phenotypic human tumor cell line profiling and predictive biomarker analysis for epigenetic inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5075. doi:10.1158/1538-7445.AM2017-5075
Abstract 5083: A novel and highly efficacious small-molecule degrader of BET-BRD proteins for the treatment of acute leukemia
Authors: Fernandez-Salas Ester , Chen Zhuo , Lin Mei , Zhou Bing , McEachern Donna , Przybranowski Sally , Kump Karson , Peterson Luke F. , Kandarpa Malathi , Hu Jiantao , Xu Fuming , Liu Liu , Bai Longchuan , Wen Bo , Sun Duxin , Talpaz Moshe , Wang Shaomeng
Published: July 2017

The bromodomain and extra-terminal domain (BET-BRD) family of lysine acetylation readers comprises BRD2, BRD3, BRD4, and BRDT that contain two conserved N-terminal bromodomains (BD1 and BD2) and a long C-terminal region containing the extraterminal (ET) protein-protein interaction domain. These proteins have key roles in the assembly of transcriptional regulatory complexes containing RNA polymerase II. BET-BRDs are new therapeutic targets for cancer treatment and several small-molecule BET-BRD inhibitors are currently in clinical development for a diverse set of cancers including leukemia. Based on the proteolysis targeting chimera (PROTAC) concept, we have developed novel and highly potent BET degraders, ZBC246 and ZBC260, with exceptional selectivity based upon a new class of BET inhibitor, ZBC11. ZBC260 effectively degrades all BET-BRD proteins at concentrations as low as 30 pM within a few hours of treatment in the RS4;11 leukemia cell line and achieves IC50 values of 50 pM in inhibition of RS4;11 cell growth. ZBC260 was tested in a panel of 10 acute leukemia cell lines generating IC50s ranging from 20 pM to 1 nM in 4-days cell growth inhibition assays. In contrast to the cytostatic effect observed with BET-BRD inhibition, degradation of BET-BRDs by ZBC246 and ZBC260 induced robust apoptosis demonstrating a differential biology between BET-BRD inhibitors and BET-BRD degraders. Significantly, ZBC260 induces rapid regression of RS4;11 xenograft tumors without overt signs of toxicity in mice. More importantly, treatment of leukemia cells obtained from 10 patients with ZBC260 demonstrated efficient degradation of BET-BRDs within 5 h at 0.3 nM, while treatment with the inhibitor (300 nM) induced up-regulation of BET-BRDs. Treatment of patients’ peripheral blasts with ZBC260 at 1 to 10 nM induced remarkable levels of apoptosis within 24 h, even in relapsed and refractory patient samples. In conclusion, ZBC260 represents a highly potent and efficacious BET-BRD degrader undergoing extensive preclinical evaluation for the treatment of acute leukemias.

Citation Format: Ester Fernandez-Salas, Zhuo Chen, Mei Lin, Bing Zhou, Donna McEachern, Sally Przybranowski, Karson Kump, Luke F. Peterson, Malathi Kandarpa, Jiantao Hu, Fuming Xu, Liu Liu, Longchuan Bai, Bo Wen, Duxin Sun, Moshe Talpaz, Shaomeng Wang. A novel and highly efficacious small-molecule degrader of BET-BRD proteins for the treatment of acute leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5083. doi:10.1158/1538-7445.AM2017-5083
Abstract 5086: Chronic radiation induced uPA expression mediates neuroblastoma cells survival
Authors: Gnanamony Manu , Fernández Karen , Libes Jaime , Lin Julian , Joseph Pushpa A. , Gondi Christopher S.
Published: July 2017

Neuroblastoma accounts for more than 15% of cancer-related deaths of children in the United States. Despite aggressive treatment regimens, the long-term survival for these children remains less than 40%. The urokinase plasminogen activator (uPA) system plays a role in several critically important biology processes, such as cell adhesion, migration, proliferation and survival. This system includes uPA, a serine protease which converts a zymogen plasminogen into the active serine protease plasmin, its surface receptor uPAR/CD57, which binds uPA with high affinity, facilitates proteolytic activation/conversion of uPA precursor (single-chain uPA, scuPA) into the active two-chain uPA protease (tcuPA) and mediates uPA-induced activation of several intracellular signaling cascades, and protects against several inhibitors, e.g. PAI-1, among others. Recent findings demonstrate that dividing/proliferating cells translocate intact scuPA to the nucleus via the nucleocytoplasmic shuttle protein nucleolin. In this study we demonstrate that both SKNBE(2) and NB1691 neuroblastoma cells when exposed to chronic radiation suppresses nMYC gene copy number and induces the overexpression of cMYC and uPA expression levels without observable change in uPA gene copy number. Increased uPA expression was accompanied with increase in the mitochondrial mass (4 fold in both NB1691 and SKNBE (2) cells). Knockdown of uPA expression in chronic radiation cells using siRNA resulted in reduced cell survival. RT-PCR analysis shows that uPA knockdown resulted in reduced expression of cMYC and its target genes cyclin A and cyclin D1. Overexpression of uPA in both SKNBE(2) and NB1691 wildtype cells resulted in increased cMYC and cyclin D1 expression but not nMYC expression suggesting that cMYC may be specifically regulated by uPA. In conclusion, our study shows that uPA plays a vital role in survival of neuroblastoma cells treated with chronic radiation by associating with cMYC to compensate for loss of nMYC expression. Our study also implies that targeting uPA might be a viable option as a combination therapy in the treatment of high-risk neuroblastoma.

Citation Format: Manu Gnanamony, Karen Fernández, Jaime Libes, Julian Lin, Pushpa A. Joseph, Christopher S. Gondi. Chronic radiation induced uPA expression mediates neuroblastoma cells survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5086. doi:10.1158/1538-7445.AM2017-5086
Abstract 5089: Antitumor activities elicited by direct intratumoral administration of a recombinant adenovirus expressing either IL-28A/IFN-λ2 or IL-29/IFN-λ1
Authors: Numasaki Muneo , Tsukamoto Hiroki , Tomioka Yoshihisa
Published: July 2017

Interleukin (IL)-28A/IFN-λ2 and IL-29/IFN-λ1 have been demonstrated to elicit direct and indirect antitumor actions. In this study, we constructed an adenovirus vector expressing either IL-28A/IFN-λ2 (AdIL-28A) or IL-29/IFN-λ1 (AdIL-29) to evaluate the therapeutic properties of intratumoral injection of recombinant adenovirus to apply for the clinical implementation of cancer gene therapy. Despite the lack of antiproliferative effect on MCA205 and B16-F10 cells, a retarded growth of established subcutaneous tumors was observed following multiple injections of either AdIL-28A or AdIL-29 when compared with AdNull. In vivo cell depletion experiments displayed that both NK cells and CD8+ T cells play a major role in AdIL-28A-mediated tumor growth suppression. A significant increase in the number of infiltrating CD8+ T cells into the tumors treated with either AdIL-28A or AdIL-29 was observed. Moreover, specific antitumor cytotoxic T lymphocyte reactivity was detected in spleen cells from animals treated with either AdIL-28A or AdIL-29. In IFN-γ deficient mice, the antitumor activity of AdIL-28A was completely impaired, indicating that IFN-γ is critically involved in the tumor growth inhibition triggered by AdIL-28A. IL-12 provided an additive antitumor effect when combined with AdIL-28A. These results indicate that AdIL-28A and AdIL-29 could be successfully utilized as an alternative cancer gene therapy.

Note: This abstract was not presented at the meeting.

Citation Format: Muneo Numasaki, Hiroki Tsukamoto, Yoshihisa Tomioka. Antitumor activities elicited by direct intratumoral administration of a recombinant adenovirus expressing either IL-28A/IFN-λ2 or IL-29/IFN-λ1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5089. doi:10.1158/1538-7445.AM2017-5089
Abstract 5097: An industrial method of manufacturing a novel RNAi anticancer drug, DFP-10825, for the treatment of peritoneal disseminated gastric cancer
Authors: Ishida Tatsuhiro , Ando Hidenori , Fukushima Masakazu , Huang Cheng-Long , Wada Hiromi
Published: July 2017

We have developed an RNAi based anticancer drug, DFP-10825 [a cationic liposome and short-hairpin RNAi molecule for thymidylate synthase (TS shRNA)] and here established an industrial method of manufacturing the formulation. The therapeutic efficacy of the DFP-10825 was examined in a MKN45 human gastric carcinoma xenograft mouse model (peritoneal disseminated gastric cancer model). The mice were received intraperitoneal injection with 5 doses of DFP-10825 or control formulation (short-hairpin RNA for luciferase) (1 mg shRNA/kg/day) once every 3 days. The treatment with DFP-10825 obviously prolonged survival time of the mice compared to control formulation and significantly suppressed the target mRNA level in the peritoneal disseminated tumors. We recently reported that DFP-10825 showed a significant therapeutic efficiency in the human malignant pleural mesothelioma orthotopic xenograft model. Accordingly, DFP-10825 may lead a clinical benefit to patients with pulmonary malignant mesothelioma or disseminated peritoneal tumor. In addition, we established an industrial method of manufacturing DFP-10825, which provides a large scale preparation. Water solution of TS shRNA was gently mixed at an optimal ratio with ethanol solution of cationic lipids, composed of DC-6-14 (a cationic lipid), DOPE and DOPC. Then, the mixture was lyophilized under optimal condition. The freeze-dried cake provided a ready for injection formulation by a one-step hydration with saline. Physicochemical properties of the obtained DFP-10825 such as particle size, zeta-potential, and size distribution were almost comparable with those of conventionally prepared DFP-10825, which prepared with conventional preparation method, mixing TS shRNA solution and cationic liposomes. Our findings in this study demonstrate that intraperitoneal chemotherapy with a simple freeze-dried RNAi medicine, DFP-10825, showing an equivalent pharmaceutical and therapeutic potency with conventionally prepared DFP-10825, meets a criteria to use for clinical settings and will have a clinical benefit to treat patients with disseminated peritoneal and pulmonary cancers.

Citation Format: Tatsuhiro Ishida, Hidenori Ando, Masakazu Fukushima, Cheng-Long Huang, Hiromi Wada. An industrial method of manufacturing a novel RNAi anticancer drug, DFP-10825, for the treatment of peritoneal disseminated gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5097. doi:10.1158/1538-7445.AM2017-5097
Abstract 510: Topoisomerase IIβ silencing increases R loops at specific genomic loci associated with an increase of γH2AX and cell cycle progression delay in human cancer cells
Authors: Delcuratolo Maria , Marinello Jessica , Capranico Giovanni
Published: July 2017

DNA topoisomerases (Top) are important players in maintaining the genome stability of cells by removing negative and positive DNA supercoils during transcription, replication and other DNA transactions. Recent evidence demonstrated that Top 1 silencing and poisoning by camptothecin affect the formation of R-loops, which are RNA/DNA hybrid structures involved in genome instability and are favored by negative supercoils of the DNA. The findings overall demonstrate that Top 1 reduces R loop levels during transcription whereas it favors R loop formation at early origins of DNA replication. Here, we address the question of whether DNA topoisomerase IIβ can also contribute to steady-state levels of R loop structures in the genome of U2OS cancer cells. We have then investigated the effects of Top2β silencing and Top2 poisons, doxorubicin and etoposide, on R loops and DNA damage in human U2OS cancer cells. Similarly to published data with camptothecin (Marinello et al., Nucleic Acids Research, 2013), Top2 poisons increase cellular R loops by IF after short treatment times and reduce them after 1 hour of treatment. The bi-phasic effect of poisons is mainly dependent on transcription. Moreover, we have determined the changes of R-loop levels by IF after Top2β silencing and the findings demonstrate that the enzyme can strongly modulate the formation of R loops as a full Top2β depletion increases nuclear R-loop levels. In addition, Top2β depletion leads to a slight increase of phosphorylation of H2AX histone along with cell cycle delay and eventually cell death. Thus, Top2β depletion can trigger genomic DNA breakage through alterations of R-loops. In order to establish which are the genomic regions of altered R loop levels, we have mapped Top2β-dependent R loop alterations by the DRIP method showing that specific genomic sites are affected by Top2β. Bioinformatic analyses of R loop maps in U2OS cells will be presented and discussed at the meeting. Altogether the findings demonstrate a critical role of Top2β in governing R loop structures in human cancer cells indicating that DNA torsional tension is a main driving factor of R loop formation and hence genome instability in cancer cells.

Citation Format: Maria Delcuratolo, Jessica Marinello, Giovanni Capranico. Topoisomerase IIβ silencing increases R loops at specific genomic loci associated with an increase of γH2AX and cell cycle progression delay in human cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 510. doi:10.1158/1538-7445.AM2017-510
Abstract 5103: Development of MJ-66 as a novel anticancer agent for the treatment of malignant gliomas
Authors: Hour Mann-Jen , Chen Tai-Lin , Gean Po-Wu , Lee Hong-Zin
Published: July 2017

Malignant glioma is the most common aggressive adult primary tumor of the central nervous system. Treatments of malignant gliomas include surgery, radiotherapy and adjuvant temozolomide (TMZ) chemotherapy. However, inherent- and acquired resistance to TMZ present major obstacles to successful treatment, and the prognosis of patients with malignant gliomas remains very poor.

MJ-66, a synthetic qunazoline compound, was identified in our study as a potent anti-proliferative agent especially on human glioma with IC50 of approximately 60 nM. In intracranial glioma xenograft model, MJ-66 (0.14 mg/kg in saline, i.p., q.d., 10 days) significantly inhibited tumor growth and increased the survival of the experimental mice, however, did not decrease the body weight of mice.

Currently, there is a profound unmet medical need for a new drug in the treatment of malignant brain tumor. The MJ-66 demonstrated superior efficacy to TMZ that are the only first-line-drug in clinical use. MJ-66 is suggested to be a promising anticancer candidate.

Note: This abstract was not presented at the meeting.

Citation Format: Mann-Jen Hour, Tai-Lin Chen, Po-Wu Gean, Hong-Zin Lee. Development of MJ-66 as a novel anticancer agent for the treatment of malignant gliomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5103. doi:10.1158/1538-7445.AM2017-5103
Abstract 5104: A novel N-mustard-quinoline conjugate is a potent agent against colorectal cancer
Authors: Chen Tai-Lin , Chen Yan-Bo , Shen Chia-Ning , Su Tsann-Long , Lee Te-Chang
Published: July 2017

Colorectal cancer (CRC) is the third leading cancer worldwide. The prognosis of patients diagnosed with CRC at the stage IV is poor with a 5-year survival rate less than 10%. Combination chemotherapy with platinum-based drugs and 5-fluorouracil (5-FU) remains the first line treatment, while the chemotherapy combined target therapeutic agents were decreasing the mortality of CRC patients. However, the efficacy of platinum-based drugs has often been limited because of the substantial risk for severe toxicities, including nephrotoxicity or neurotoxicity. Therefore, there is an unmet need to develop a novel drug for improving the treatment of advance colorectal cancer. We have previously designed and synthesized several series of DNA-directed alkylating agents with potent antitumor activity. Recently, one derivative of N-mustard-quinoline conjugates with hydrazinecarboxamide as a linker, designated as SL-1, was selected to explore its anti-colorectal cancer (CRC) activity because it was highly cytotoxic to a panel of CRC cell lines in culture. Flow cytometric analysis showed that treatment of CRC cells with SL-1 induced G2/M arrest and triggered apoptosis. We further demonstrated that SL-1 preferentially targeted on the selective guanine sequence by sequencing gel electrophoresis. By the aid of comet assay, SL-1 in combination with 5-FU resulted in increased DNA tail moments and induced more DNA damage. Evaluation of the potency of antitumor activity both in culture and in xenograft models, we observed that SL-1 was more potent than 5-FU and oxaliplatin against SW620, RKO and RKO-E6 (an oxaliplatin resistant-subline) cells. Notable, SL-1+5-FU were more efficacious than oxalipatin+5-FU in suppression the growth of RKO or SW620 in xenograft models. We further found that SL-1 enhanced 5-FU and induced more apoptosis by c-caspase 3 and TUNNEL staining. Additionally, histopathological examination showed that SL-1 by itself has no obvious kidney and liver toxicity in ICR mice comparing to cisplatin, oxaliplatin or 5-FU. Taken together, SL-1 alone or in combination with 5-FU may warrant our further development as a potential antitumor agent for the treatment of CRC patients.

Citation Format: Tai-Lin Chen, Yan-Bo Chen, Chia-Ning Shen, Tsann-Long Su, Te-Chang Lee. A novel N-mustard-quinoline conjugate is a potent agent against colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5104. doi:10.1158/1538-7445.AM2017-5104
Abstract 5105: Structure-activity relationship study of pyrrolo[2,3- d]pyrimidines as microtubule targeting antitumor agents
Authors: Gangjee Aleem , Quadery Tasdique M. , Mooberry Susan L.
Published: July 2017

Microtubules are cytoskeleton protein polymers composed of αß-tubulin heterodimers which play pivotal roles in cell division, cellular transport and cell signaling. Interfering with microtubule dynamics is a well-established strategy for the treatment of cancer. However, clinical utility of microtubule targeting agents (MTAs) in cancer chemotherapy is often limited by the emergence of drug resistance. Expression of P-glycoprotein (Pgp) or the βIII isotype of tubulin, are two clinically established drug resistance mechanisms. We previously reported that compound AG346 binds to the colchicine site of tubulin and has microtubule depolymerization activity in A-10 cells (EC50 = 365 nM). Molecular modeling and docking studies of this compound in the X-ray crystal structure of tubulin (PDB: 402B) suggested that the pyrrole NH can be substituted with alkyl chains of optimal length to obtain an altered binding conformation which can facilitate interaction with the hydrophobic pocket formed by amino acid Val181 of side chain A, and Lys352 and Thr314 of side chain B. We designed and synthesized a variety of pyrrole NH-alkyl substituted compounds and determined that the compound AG473 shows a 43-fold improved microtubule depolymerization potency (EC50 = 8.4 nM) in A-10 cells, compared to the lead compound. In addition, this compound inhibited the growth of human melanoma cancer cell line MDA-MB-435 in culture with an IC50 of 7.2 nM, which is a 5-fold improvement in potency over the parent compound. These analogs also circumvented resistance mediated by Pgp and βIII-tubulin. These attributes provide the impetus for further preclinical development of these compounds as microtubule targeted antitumor agents in vivo and these studies are currently underway.

Citation Format: Aleem Gangjee, Tasdique M. Quadery, Susan L. Mooberry. Structure-activity relationship study of pyrrolo[2,3- d]pyrimidines as microtubule targeting antitumor agents [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5105. doi:10.1158/1538-7445.AM2017-5105
Abstract 5109: Beyond malaria: Second career of artesunate as cancer drug
Authors: Efferth Thomas A.
Published: July 2017

More than a decade ago, we initiated a research program on the molecular pharmacology of phytochemicals derived from Chinese medicinal herbs. Bioactive plant extracts have been fractionated by chromatographic techniques. We isolated bioactive compounds and elucidated their chemical structures by nuclear magnetic resonance and mass spectrometry. A promising compounds was artemisinin from Artemisia annua L. and its semisynthetic compound artesunate. Artemisinin and artesunate are anti-malarial drugs. Our data indicated profound activity against cancer cells, but also against various viruses, Schistosoma, Trypanosoma, and even plant crown gall tumors. To elucidate the molecular mode of actions against cancer, we applied molecular biological and pharmacogenomic approaches in vitro and in vivo. Different signaling pathways were identified not only in cancer cells but also in cells infected with viruses, e.g. HCMV, HSV1 and others. To translate the experimental results in cell lines and animals to the bedside, we report on the compassionate use of artesunate in single cancer patients as well as on our efforts to initiate several clinical phase I/II trials in veterinary tumors as well as in human cervix or colorectal carcinoma. These pilot studies indeed indicate that artesunate is not only useful as antimalarial drug, but also exerts activity against cancer and viral diseases. Clinical results will also be presented that not only artesunate as semisynthetic chemical derivative of artemisinin, but also herbal extracts from Artemisia annua are active in veterinary and human tumor patients. Artesunate represents an illustrative example for the therapeutic potential of medicinal herbs and drugs derived from traditional Chinese medicine.References 1. Efferth et al.: Journal of Molecular Medicine 2002;80:233-42. 2. Efferth: Drug Resistance Updates 2005;8:85-97. 3. Efferth et al.: Molecular Cancer Therapeutics 2008;7:152-61. 4. Li et al.: Cancer Research 2008;68:4347-51. 5. Shapira et al.: Clinical Infectious Diseases 2008;46:1455-7. 6. Efferth et al.: Clinical Infectious Diseases 2008;47:804-11. 7. Krishna et al.: Ebiomedicine 2014;2:82-90. 8. Saeed et al.: Pharmacological Research 2016;110:216-226.

Citation Format: Thomas A. Efferth. Beyond malaria: Second career of artesunate as cancer drug [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5109. doi:10.1158/1538-7445.AM2017-5109
Abstract 511: Chromosome instability mechanisms in breast cancer among African Americans and Caucasians
Authors: Thaiparambil Jose , Mansour Oula , Haley Susan , EL-Zein Randa
Published: July 2017

Significant disparities exist in breast cancer among African Americans (AAs) women compared to Caucasians. Even though AAs are less likely to undergo regular mammograms due to access barriers, it is unlikely that differences in screen based detection accounts entirely for outcome disparities. While the current guidelines recommend women begin mammography at age 40 or 50, AA women may be at increased risk for delayed diagnosis since >10% of breast cancer cases in AAs are diagnosed in women younger <40 years compared with 5% of Caucasians. This highlights the importance of defining optimal ethnically appropriate age of screening especially given the disproportionate mortality among younger black women patients. Therefore, an alternative approach using biomarkers should be considered in order to develop ethnically appropriate measures for detecting the disease in the earliest stages. Genomic instability has long been recognized as a major driver of carcinogenesis occurring early on and increasing in complexity with disease progression. However, to date little is known about the underlying mechanisms associated with such instability among the different ethnic groups. In this study we hypothesized that alterations in telomere length and dysfunctional centrosome apparatus are underlying mechanisms associated with ethnic-based genomic instability. We investigated alterations in centrosome apparatus, misaligned chromosomes and spindle angle orientation in AA [HCC 70] and Caucasian [HCC1395] cell lines using confocal microscopy. Our results demonstrated significant differences in structural (30%) and numerical alterations (60%) in centrosome apparatus between AA and Caucasian cell lines (p<0.001). We also noticed a significant increase of misaligned chromosomes (35%) and (68 %) in the metaphase of Caucasians and AA’s respectively (p<0.002). In addition, a significant difference in spindle orientation was observed among AAs and Caucasians suggesting that mitotic phase abnormalities may be, in part, responsible for the underlying genetic instabilities associated with breast cancer. Using ingenuity pathway analysis, we compared gene expressions in both AAs and Caucasian cell lines and observed a significant change in telomerase signaling expression between the cell lines, suggesting a role for telomere lengths in the generation of genetic instability. Together, identifying the specific mechanisms that drive tumorigenesis may provide opportunities for future targeted prevention efforts among high risk cohorts. (This work is supported in part by CA189240)

Citation Format: Jose Thaiparambil, Oula Mansour, Susan Haley, Randa EL-Zein. Chromosome instability mechanisms in breast cancer among African Americans and Caucasians [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 511. doi:10.1158/1538-7445.AM2017-511