bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1764: Changes in apoptotic gene expression in human leukemia cells treated with the anticancer drug Laromustine
Authors: Kaye Emily G. , Gross Kayla M. , Bellairs Jo , Dale Lindsay E. , Gallagher Catherine , Sher Emily V. , Robinson Kristen N. , Rice Kevin P.
Published: August 2012

Laromustine is an experimental anticancer drug that has been tested in the clinic against acute myelogenous leukemia and glioblastoma multiforme. In the body, Laromustine's cytotoxic effects are a result of two cogenerated electrophiles, a 2-chloroethylating species that can cause lethal DNA damage and methylisocyanate, which carbamoylates sulfydryl groups in proteins. The mechanism by which Laromustine kills cancer cells likely includes apoptosis. In order to further investigate this phenomenon, the extent to which Laromustine affects cultured human promyelocytic (HL-60) cells’ use of 88 genes relating to this cell death pathway was examined using quantitative real-time reverse transcriptase PCR. Cultured cells were treated with drug for six hours before harvesting mRNA for analysis. The expression of a subset of the tested genes emerged as significantly different in cells treated with Laromustine as compared to control cells. Included among these genes are: TNFRSF10D, TNFRSF17, CD40LG, TRAF4, and BAG3. The expression of BAG3, whose encoded protein is also known as CAIR-1 or Bis, was increased by more than two orders of magnitude in HL-60 cells exposed to Laromustine. By forming a complex with heat shock protein 70 (Hsp70), it is thought that BAG3 promotes anti-apoptotic activity by interfering with protein chaperoning and cytochrome-c release. It is possible that a pathway involving BAG3 could be a mechanism by which cells attempt to counter the effects of Laromustine exposure. Other apoptotic markers are also reported, including activation of caspase-3 and caspase-7, cleavage of poly(ADP-ribose) polymerase, and staining of annexin V.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1764. doi:1538-7445.AM2012-1764
Abstract 1765: Potent antitumor BO-1922, derivative of indolizino[6,7-b]indole, against human colon, lung, and pancreatic cancers in xenograft model
Authors: Su Tsann-Long , Tala Satishkumar , Chaniyara Ravi , Chen Chi-Wei , Lee Te-Chang , Chou Ting-Chao
Published: August 2012

Our previously studies demonstrated that bis(hydroxymethyl)cyclopenta[a]indenes, which bear a bis(hydroxymethyl)pyrrolidine pharmacophore, exhibited potent antitumor activity in inhibiting various human tumor xenografts. We also demonstrated that these agents showed strong synergistic therapeutic efficacy in combination with arsenic trioxide (ATO) against acquire or inherited MDR human tumors in xenograft model (human lung H460 or bladder cancer resistant to cisplatin NTUB1/P). Based on these findings, we have recently synthesized a series of 1,2-bis(hydroxymethyl)indolizino[6,7-b]indols and their bis(alkylcarbomate) derivatives for antitumor evaluation. These new agents were easily prepared starting from L-tryptophane in 5 steps. The in vitro cytotoxicity studies showed that the newly synthesized derivatives possessed a broad spectrum of anticancer activity against human lymphoblastic leukemia and various solid tumor cell growth in vitro. Of these analogues, 1,2-bis(hydroxymethyl)-3,6-dimethylindolizino[6,7-b]indole (BO-1922) was selected for further antitumor studies. We found that this agent displayed potent therapeutic efficacy against human colon cancer HT-29, lung adenocarcinoma lung A549, and primary pancreatic adenocarcinoma BX-PC3 xenografts. The antitumor activity of BO-1922 was compared with Irinotecan (CPT-11) in animal xenograft model. It revealed that BO-1922 is more potent than CPT-11 against human colon cancer HT-29 xenograft; complete tumor suppression on day 20 after mice treated with BO-1922 (25 mg/kg, QD×4, intravenous injection), while, tumor relapsed on day 9 after mice treated with CPT-11 (60 mg/kg, Q2D×3, iv. inj.). Under the same drug administration conditions (dosage and schedule), BO-1922 is as potent as CPT-11 against lung cancer A549 and pancreatic cancer BX-PC3 xenografts. Although the bodyweight of all drug treated mice recovered after ceasing of treatment, it was found that BO-1922 is less toxic to the host based on the average body-weight loss. The alkaline agarose gel shifting assay showed that this agent is able to induce DNA double strand cross-linking. Flow cytomeric assay revealed that BO-1922 induced cell cycle arrest at G1/S phase in non-small cell lung carcinomas H1299. These results suggest that BO-1922 is a promising candidate for clinical application and is currently undergoing preclinical studies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1765. doi:1538-7445.AM2012-1765
Abstract 1768: Enhanced cytotoxicity and apoptosis of cisplatin by p53-reactivating small molecule RITA in head and neck cancer
Authors: Ryu Chang Hwan , Ryu In Sun , Park Gi Cheol , Choi Jun Young , Park Jung Je , Roh Jong-Lyel
Published: August 2012

Overexpression of MDM2 inactivates the tumor suppressive function of p53 and a small-molecule reactivation of p53 and induction of tumor cell apoptosis (RITA) disrupts the MDM2-p53 interaction. We evaluated whether the restoration of p53 function by RITA enhances cytotoxicity and apoptosis of cisplatin in head-and-neck cancer (HNC). RITA was tested in four human HNC cell lines differing in TP53 status. In vitro and in vivo growth suppression, cell cycle arrest, apoptosis after RITA treatment individually or in combination with cisplatin were assessed. RITA induced prominent accumulation and reactivation of p53 in a wild-type TP53-bearing tumor cell line. RITA showed maximal growth suppression in tumor cells showing MDM2-dependent p53 degradation. RITA upregulated p21, BAX, and cleavage of caspase-3 but the p53-dependent apoptosis was blocked by pifithrin-μ. Strong induction of apoptosis rather than G2-phase arrest was observed along with increased dose of RITA. In combination therapy, RITA enhanced the growth inhibition and apoptosis of HNC by cisplatin in vitro and in vivo. Our data suggest that the restoration of p53 tumor suppressive function by RITA enhances cytotoxicity and apoptosis of cisplatin, which may offer an attractive strategy for treating HNC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1768. doi:1538-7445.AM2012-1768
Abstract 1777: Antitumor effect of PM01183 in a human pancreatic adenocarcinoma orthotopic model
Authors: Cespedes M Virtudes , Guillén M José , Mangues Ramon , Gallardo Alberto , Alamo P , Casanova I , Cuevas Carmen , Aviles Pablo
Published: August 2012

Background: Pancreatic ductal adenocarcinoma is the fourth cause of death in the Western world and, despite the advances in cancer therapies, limited therapeutic benefit (palliating symptoms, better quality of life) may be offered to patients: the overall 5-year survival rate remains < 5%. PM01183 is a new synthetic tetrahydroisoquinoline alkaloid that binds to selected DNA sequences and promotes apoptosis by inducing double-strand breaks at nanomolar concentrations. Confirmed antitumor clinical activity was observed at the recommended dose level of PM01183 in the first-in-human Phase I study in patients with advanced/refractory solid tumors. Currently, PM01183 clinical development includes Phase II single agent trials (pancreas, breast and platinum-resistant/refractory ovarian) as well as Phase I trials in combination (with doxorubicin and gemcitabine) and in advanced acute leukemia. The in vivo antitumor activity of PM01183 was investigated in the human NP9 pancreatic orthotopic model Material and Methods: Athymic nude female mice were orthotopically implanted with luciferase-expressing NP9Luc tumors. Fifteen days after implantation, tumor bearing mice were randomly allocated (N=12 /group) into either PM01183 (at 0.18 mg/kg), gemcitabine (at 180 mg/kg) or control (placebo) groups. Treatments were initiated (Day 0) and administered for 3 consecutive weeks (q7dx3; days 0, 7, 14). Response to treatment was assessed on Day 7 (short-term) by bioluminescence imaging (BLI), and by survival evaluation (long-term): mice were sacrificed when their tumor diameters reached ca. 2 cm or when they became moribund. Statistical differences between groups were determined using two-tailed Mann-Whitney U test and by analysis of the Kaplan-Meier curve. Results: Results showed that PM01183 (q7dx3 at 0.18 mg/kg) induced statistically significant tumor reduction compared to placebo-treated animals by BLI analysis (P= 0.006) on Day 7. Reductions in tumor size were associated to changes in survival median times: PM01183 treatment significantly increased the survival time compared to either gemcitabine- (P= 0.003) or placebo-treated (P= 0.0001) animals. Conclusion: PM01183 demonstrated in vivo antitumor activity in a human pancreatic orthotopic model (NP9), as reflected by metabolic, size-related and survival rate.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1777. doi:1538-7445.AM2012-1777
Abstract 1778: Antitumor effect of PM01183 in a patient-derived Cisplatin-sensitive and -resistant serous epithelial ovarian orthotopic tumor model
Authors: Villanueva Alberto , Vidal August , Guillén M José , Salazar Ramón , Cuevas Carmen , Aviles Pablo M.
Published: August 2012

Background: Epithelial ovarian cancer (EOC) is the leading cause of deaths in women diagnosed with gynecologic malignancies. The low survival rate is due to its advanced-stage diagnosis and either intrinsic or acquired resistance to standard platinum-based chemotherapy. As such, the development of effective innovative therapeutic strategies to overcoming cisplatin resistance remains a high priority. PM01183 is a new alkaloid that binds to the minor groove of DNA and induces delayed progression of S phase and cycle arrest in G2/M. Currently, PM01183 clinical development includes Phase II single agent trials (pancreas, breast and platinum-resistant/refractory ovarian) as well as Phase I trials in combination (with doxorubicin and gemcitabine) and in advanced acute leukemia. Here, we report that PM01183 is effective in the treatment of new developed orthotopic subject-derived epithelial serous ovarian tumor grafts Cisplatin-sensitive and-resitan models. Material and Methods: Athymic nude female mice were orthotopically engrafted with derived-fragments of OVAX1 (Cisplatin-Sensitive) or OVAX1R (Cisplatin-Resistant) tumors, and when tumors reached homogeneous palpable size were randomly allocated into the treatment groups (N = 8-12/group): i) Placebo; ii) PM01183 (0.18 mg/kg); iii) Cisplatin (3.5 mg/kg); and, iv) PM01183 plus Cisplatin (0.18+3.5 mg/kg). Drugs were intravenously administered once per week for 3 consecutive weeks (days 0, 7 and 14) and seven days after the last dose (Day 21), animals were sacrificed, their ovaries dissected-free and weighed. Statistical differences between groups were determined using two-tailed Mann-Whitney U test. Results: On day 21, OVA1X tumors experienced reductions of 95.3, 88.3 and 87.2% following the treatment with Cisplatin, PM01183 and PM01183+Cisplatin, respectively. The results obtained for OVA1XR tumor demonstrated tumor weight reductions of 48.2, 93.6 and 96.7% for Cisplatin, PM01183 and PM01183+Cisplatin treatments, respectively. Single therapy with PM01183 induced a statistically significant better response than Cisplatin (P=0.003). Also, PM01183+Cisplatin treatment increased the activity following Cisplatin (P=0.002) or PM01183 (P=0.022) administration as single agents, suggesting thus a synergistic effect of both compounds. Conclusions: PM01183 demonstrated strong in vivo antitumor activity in pure patient-derived Cisplatin-sensitive and Cisplatin-resistant serous epithelial ovarian orthotopic tumor models.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1778. doi:1538-7445.AM2012-1778
Abstract 178: A novel mechanism of hypoxia mediated cell cycle dysregulation: miR-134 targets CDKN2A
Authors: Rupaimoole Rajesha , Pecot Chad V. , Wu Sherry Y. , Bottsford-Miller Justin , Ivan Cristina , Coleman Robert L. , Zhang Wei , Sood Anil K.
Published: August 2012

Background: Among the various factors in the tumor microenvironment influencing tumor growth and progression, hypoxia plays a key role in promoting tumor growth and resistance to therapeutics. However, a mechanistic understanding of the complicated biological interplay between the stromal framework (hypoxia context) and malignant regions of the same tumor remains an unmet need. In this study, we examined the molecular networks of interplay between hypoxia and miRNA deregulation. Study methods and results: In this study, we profiled the expression of miRNAs in ovarian cancer cells upon exposure to hypoxia (1% O2) using microRNA microarray (Agilent v14). We found that significant numbers of miRNAs are dysregulated. In A2780 and Skov3 ovarian cancer cells, using a statistical cutoff (p<0.05 and fold change >1.5), normoxia v/s short term (48hrs) and long term (6 days) hypoxia exposure, we found that 23 MicroRNAs were down regulated whereas 11 were upregulated. Among these, miR-134 validated to be overexpressed under hypoxia by 3 - 4 fold in A2780 and SKOV3 ovarian cancer cells consistently across the short and long term time points. Upon bioinformatics analyses for target sites of commonly up regulated miRNAs, we found that miR-134 was predicted to target CDKN2A (Miranda, microT, PITA, RNA22, TargetScan). CDKN2A is a protein involved in cell cycle control, precisely regulating cell cycle G1 control through complexing with CDK4. Considering the fact that CDKN2A expression is lost in several cancer types and its correlation with the aggressive tumor growth, we checked expression of CDKN2A under hypoxic conditions. CDKN2A was down-regulated under hypoxia by 55% and was inversely correlated with miR-134 expression in cell lines tested. Upon transfection of A2780 ovarian cancer cells with miR-134 microRNA mimics, we observed consistent down regulation of CDKN2A by >40%, compared to scrambled control transfected cells. This further proves the role of miR-134 in deregulating CDKN2A expression in ovarian cancer. Discussion: This study provides new insights into miRNA-134's role in regulating cell cycle in the context of hypoxia mediated signaling. These findings may have implications for new approaches targeted at curbing the deleterious effects of hypoxia on tumor growth.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 178. doi:1538-7445.AM2012-178
Abstract 1781: LGI147, a novel small-molecule inhibitor of the mitotic kinesin Eg5, shows anticancer effects in human hepatocellular carcinoma
Authors: Lin Zhong-Zhe , Huang Chi-Ying F. , Pan Hung-Wei , Sun Nai-Yun , Hsiao Li-Pin , Hsu Hey-Chi , Cheng Ann-Lii
Published: August 2012

Background: Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide, for which novel therapies are urgently needed. Recently, the inhibitors of mitotic kinesin Eg5 emerge as potential anticancer therapeutics. The aim of this study was to investigate the anticancer activity of a potent Eg5 inhibitor, LGI147, as targeted therapy of HCC. Method: Antiproliferative effect of LGI147 (kindly provided by Novartis), a highly selective inhibitor of mitotic kinesin Eg5, was tested in three liver cancer cell lines (Hep3B, HepG2, PLC/PRF/5). Cell viability was measured by trypan blue exclusion assay. Mitotic perturbation was visualized by immunofluorescence staining and confocal microscopy. Cell cycle profiles were assessed by flow cytometry. Apoptosis was estimated by analysis of sub-G1 fraction, Annexin V expression and immunoblotting of apoptotic proteins. Results: LGI147 induced a dose-dependent growth inhibition with IC50s ranging from 43.47 pM to 53.59 pM in the tested HCC cell lines. LGI147 treatment for 24 hours led to inhibition of Eg5 activity and induction of abnormal mitosis in a dose-dependent manner. Polypolar spindle defects in mitosis, disturbed cell cycle progression, and induction of apoptosis were observed in the HCC cells treated with LGI147. Conclusions: These findings show that LGI147 has potent anticancer effects in human HCC. Inhibitors of mitotic kinesin Eg5 may deserve further exploration as molecular targeted agents against HCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1781. doi:1538-7445.AM2012-1781
Abstract 1786: Characterization of the inhibitory capacity of EMD1214063, a novel small molecule inhibitor of the MET hepatocyte growth factor receptor on a panel of MET mutated variants
Authors: Zimmer Yitzhak , Pochon Benoît , Stroka Deborah M. , Aebersold Daniel M. , Blaukat Andree , Bladt Friedhelm , Medová Michaela
Published: August 2012

The receptor tyrosine kinase MET is a prime target in clinical oncology because of its aberrant activation in a broad spectrum of malignancies. Here, we tested the anti tumor activity of a novel MET inhibitor, EMD1214063, on cancer cells overexpressing MET and in eight transfected NIH3T3 cell lines expressing different activating mutants of the MET kinase. Our results demonstrate a dose-dependent decrease in MET autophosphorylation in both MET overexpressing cell lines (IC50 of 20nM and 30nM for GTL-16 and H1993, respectively) and in six of the eight cell lines transfected with MET activating mutants (IC50 5-18nM). Blockade of MET by EMD1214063 resulted in reduced downstream activation of AKT, ERK and PLCγ. In contrast, lack of MET inhibition by EMD1214063 in the resistant Y1248H and L1213V MET mutants was associated with high levels of AKT, ERK and PLCγ activation. In all sensitive mutants, EMD1214063 attenuated MET-dependent cellular proliferation and significantly altered cell cycle distribution profiles (increase in G1 and concomitant decrease of S phase). Furthermore, EMD1214063 strongly affected MET-driven biological functions, such as cellular morphology, motility and anchorage-independent growth. To assess the in vivo efficacy of EMD1214063, we established tumor transplantation models exploiting NIH3T3 cells expressing the H1112L or L1213V MET mutants, respectively sensitive and resistant, to in vitro treatment with EMD1214063. Animals bearing H1112L or L1213V tumors were randomized for the treatment with EMD1214063 (50mg/kg/day) or vehicle only. Remarkably, 5 days of treatment with EMD1214063 induced complete regression of sensitive H1112L tumors, while tumor growth remained unaffected in mice injected with resistant L1213V tumor cells and in vehicle-treated mice. Taken together, these results underscore the concordance between in vitro and in vivo effects of EMD1214063 and strongly support its efficacy in inhibiting MET-driven tumor cell growth.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1786. doi:1538-7445.AM2012-1786
Abstract 1793: Imatinib treatment for gefitinib-resistant lung cancer harboring epidermal growth factor receptor mutation
Authors: Fukuhara Tatsuro , Treda Cezary Jan , Sakakibara Tomohiro , Inoue Akira , Ebina Masahito , Nukiwa Toshihiro
Published: August 2012

Background: Epidermal growth factor receptor (EGFR) mutant-lung cancer showed significant response to EGFR-tyrosine kinase inhibitor (TKI) treatment, however, treatment failure due to acquired resistance to EGFR-TKI become important clinical problem for long-term survivors. Our purpose is to develop the new treatment for EGFR-TKI resistance. Methods: We used PC9 human lung cancer cell line harboring EGFR exon 19 deletion (E746-A750). When PC9 cells were cultured with 2 μM gefitinib solved in dimethylsulfoxide (DMSO), most of cells died and surviving cells proliferated slowly. By 90 days-culture with 2 μM gefitinib, PC9 gefitinib resistant cells (PC9GR90) were established and confirmed that EGFR-T790M resistance-related mutation was negative. In PC9GR90, an immunoblot assay revealed MET (hepatocyte growth factor receptor) amplification and alternative EGFR3 tyrosine phosphorylation, which is known as one of the major resistant mechanisms. Results: When PC9GR90 were cultured with 2 μM gefitinib and one inhibitor (histone deacetylase inhibitor, insulin-like growth factor receptor inhibitor, gamma secretase inhibitor, or imatinib known as bcr-abl, c-kit and platelet derived growth factor receptor inhibitor), 10 μM imatinib treatment with gefitinib showed significant suppression of cell growth. The ratio of cell count of 2 μM gefitinib/DMSO and 10 μM imatinib with 2 μM gefitinib/DMSO were 91.7% and 6.0%, respectively. When imatinib concentration was decreased to 1 μM known as appropriate concentration for chronic myeloid leukemia treatment, there was no effect. The imatinib effect was not shown for NCI-H1975 lung cancer cells harboring both EGFR-L858R and -T790M. By immunoblot, we found EGFR3 phosphorylation was inhibited by imatinib treatment. Conclusion: It is the first report that imatinib inhibits EGFR3 phophorylation and induce cell death in the EGFR-TKI resistant lung cancer with MET amplification and EGFR3 phophorylation. Although further studies are required to apply this strategy to a clinical setting, we demonstrate a hopeful data to conquer one important resistant mechanism.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1793. doi:1538-7445.AM2012-1793
Abstract 1798: Identification of pan-Trk inhibitors for the treatment of Trk-driven cancers
Authors: Bouhana Karyn S. , Gomez Anna M. , Alford Taylor J. , Woessner Richard , von Carlowitz Ira , Lee Patrice A. , Andrews Steven D. , Winski Shannon L.
Published: August 2012

The Trk family of genes, which includes trkA/NTRK1, trkB/NTRK2 and trkC/NTRK3, encode the tyrosine kinase receptors for the neurotrophin family of nerve growth factors. Deregulated kinase activity of trk family members is associated with human cancer. Oncogenic translocations involving trkC kinase domain have been identified in AML, salivary gland carcinoma, adult secretory breast cancer, congenital fibrosarcoma, and pediatric nephroma. Oncogenic trkA translocations have been reported in papillary thyroid and colorectal cancers. We have identified orally bioavailable, potent and selective ATP-competitive inhibitors of the trk family of receptor tyrosine kinases and are developing these for the treatment of Trk-driven cancers. AR523 is a pan-Trk inhibitor which demonstrates similar activity against TrkA, TrkB and TrkC receptors in a cell based assay (IC50 ∼10 nM). In a screen at 1 µM against a panel of 230 kinases, AR523 inhibited only three additional kinases more than 50% (TNK2, Bmx and Txk). In cell culture AR523 was found to exhibit selective anti-proliferative activity toward the cell line KM12, which harbors the oncogenic TPM-TrkA translocation, while exhibiting no activity toward HT29, a cell line with no trk gene rearrangements. Studies in mice engrafted with KM12 tumors revealed significant anti-tumor and pharmacodynamic activity of AR523. Administration of a single oral dose of AR523 reduced tyrosine phosphorylation of TrkA by nearly 80% at twelve hours after dosing. Parallel analysis of Akt and Erk revealed reduced phosphorylation of these downstream effectors. Administration of 100 mg AR523 daily for two weeks to mice with KM12 xenografts produced mean tumor growth inhibition of 86% and a mean of 42% tumor regression. AR523 exhibits dose dependent inhibition of tumor growth at 10 and 30 mg/kg in KM12 xenografts with 54 and 72% TGI respectively. AR523 was well-tolerated, causing no weight loss or deaths compared with vehicle control. Increasingly Trk mutations have been shown to be activating and the use of Trk inhibitors may provide a new therapeutic strategy for targeted treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1798. doi:1538-7445.AM2012-1798
Abstract 1801: Lyn and Fyn inhibition as a potential novel treatment for glioblastoma multiforme
Authors: Prakash Om , Wilk Anna , Vashistha Himanshu , Lin Shih-Chao , Aregbezola Rita , Levitt Daniel , Reiss Krzysztof
Published: August 2012

Background: Glioblastoma multiforme (GBM) is the most malignant and most aggressive of the adult brain tumors. The current therapies including surgical resection, radiation therapy and chemotherapy remain palliative with median survival of 14 months. Two of the Src family kinases, Lyn and Fyn, have been reported to be involved in the pathogenesis and aggressiveness of GBM in clinical and laboratory research studies. Specific Aims: We evaluated the effect of Lyn and Fyn kinase inhibition on GBM cell motility and survival by utilizing a clinically relevant Lyn and Fyn kinase inhibitor, bafetinib or formerly INNO-406 (CytRx Corporation, Los Angeles, CA). Bafetinib, a dual Bcr-Abl and Lyn tyrosine kinase inhibitor has been shown to inhibit 4 of the 79 tyrosine kinases, namely Abl, Abl-related gene (Arg), and two of the Src family kinases Lyn and Fyn but not Src. Methodology: We used Lyn- expressing T98G and Fyn-expressing U87MG human GBM cell lines. The effect of bafetinib on the motile activity was evaluated by the wound healing, single cell trajectory, and the trans-well assays. The cell survival and mitochondrial membrane potential were evaluated by Guava Easycyte *HT flowcytometer (Millipore) in cells treated with bafetinib for 24 h. Further, we examined the effect of bafetinib on cell survival in combination with temozolomide (TMZ), a widely used chemotherapy agent for the management of GBM, in U87MG cells using microplate MTT assay. Results: At 16 h post-wounding, bafetinib at 5 µM concentration inhibited the motility of T98G and U87MG cells by 33% (P < 0.01) and 24% (P < 0.01), respectively. This significant inhibition in the wound healing was accompanied by the inhibition of total length of cell trajectory, total length of the final cell displacement and average speed of cell locomotion. Bafetinib induced 62% mitochondrial membrane depolarization in U87MG cells and 20% depolarization in T98G cells. Higher depolarization levels in U87MG cells correlated with higher apoptosis (49.4%). Treatment of U87MG cells for 4 days with a combination of bafetinib and TMZ reduced the IC50 values as much as 60% and 80%, respectively, compared to the values obtained with each of the drugs, suggesting that bafetinib has the potential to improve the therapeutic effect of TMZ in GBM cells. Conclusion: Our preliminary findings suggest that targeting Lyn and Fyn kinase pathways with bafetinib may have a therapeutic potential for GBM. Our in vivo studies in mouse models with bafetinib will further establish the feasibility of targeting Lyn and Fyn kinases as possible therapeutic targets in GBM.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1801. doi:1538-7445.AM2012-1801
Abstract 1802: IGF-1R inhibition as a novel therapeutic strategy for small cell lung cancer
Authors: Zinn Rebekah L. , Rudin Charles M.
Published: August 2012

Small cell lung cancer (SCLC), which makes up approximately 15% of all lung cancer cases, is characterized by short responses to standard chemotherapy and a median survival duration of less than one year. New therapies are critically needed to improve the outcome for patients with this disease. Signaling through the insulin-like growth factor 1 receptor (IGF-1R) leads to activation of the PI3K-AKT and MAP kinase pathways, resulting in inhibition of apoptosis and stimulation of proliferation. The IGF-1R pathway is commonly upregulated in SCLC. Recent evidence suggests that inhibition of IGF-1R may be a potential treatment strategy for many types of cancer, including SCLC. We sought to characterize the relative sensitivity of SCLC cell lines to a novel IGF-1R tyrosine kinase inhibitor, OSI-906, identify predictive biomarkers of sensitivity, and evaluate the efficacy of OSI-906 in vivo on SCLC cell line xenografts and primary patient xenografts as a single agent and in combination with etoposide. We assessed the sensitivity of a panel of 16 SCLC cell lines to OSI-906. Approximately one third of these cell lines were highly sensitive to OSI-906, with an IC50 < 1 μM, as measured by the MTS cell proliferation assay. Treatment with increasing doses of OSI-906 resulted in dose dependent inhibition of phosphorylated IGF-1R and phosphorylated AKT in both sensitive and resistant cell lines, suggesting that while the IGF-1R signaling pathway is intact in resistant cell lines, they rely on other signaling pathways for survival. Expression profiling of our panel of cell lines for components of the IGF-1R pathway including IGF-1R, insulin receptor (IR), and receptor ligands (IGF-1 & IGF-II) did not suggest a simple correlation between receptor/ligand expression and sensitivity to OSI-906. Interestingly, OSI-906 sensitive cell lines appear to express lower levels of phosphorylated ERK at baseline compared to resistant cell lines. In addition, phospho-ERK is not inhibited by OSI-906 and is upregulated in response to treatment in some resistant cell lines. We tested the efficacy of OSI-906 alone and in combination with etoposide in vivo using an H187 cell line xenograft model and two primary patient xenograft models. OSI-906 alone resulted in approximately 50% tumor growth inhibition in H187 xenografts and 30% inhibition in the primary patient xenograft models compared to mock treated animals. There was no evident synergy with etoposide in any of these models. Our preliminary data support that IGF-1R inhibition may be a viable treatment strategy for a subset of SCLC. Identification of the subset that will respond to this therapy may be more complex than screening for expression IGF-1R pathway components.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1802. doi:1538-7445.AM2012-1802
Abstract 1806: The indolocarbazole derivative Go6976 suppresses the growth of leukemia cells with FLT3 mutations
Authors: Yoshida Akira , Zokumasu Kouichi , Ueda Takanori
Published: August 2012

The FMS-like tyrosine kinase 3 (FLT3) is a class III receptor tyrosine kinase involved in hematopoietic progenitor cell development. Mutations of FLT3 have been reported in about a third of patients with acute myeloid leukemia (AML), and inhibitors of FLT3 are of clinical interest. Go6976 is an indolocarbazole inhibitor of the calcium-dependent isozymes of protein kinase C (PKC). In the present study, we demonstrated that Go6976 possesses a potent inhibitory activity against recombinant FLT3 using in vitro kinase assay. We also showed that Go6976 is a potent inhibitor of Aurora-B kinase. Go6976 significantly inhibited proliferation of leukemia MV4-11 cells having FLT3-internal tandem duplication (ITD). Cell growth inhibition by Go6976 occurred in parallel with the drug inhibiting FLT3 and its downstream signal pathways such as extracellular signal-regulated kinase1/2, STAT-5 and p38. The FLT3-independent myeloid leukemia HL-60 and U937 showed strong resistance to Go6976 treatment. Induction of massive apoptosis were observed upon treatment with Go6976 in MV4-11 cells associated with significant down-regulation with Survivin and Mcl-1 protein. Interestingly, Go6976 treatment also inhibited Survivin phosphorylation on Thr34, which is critical for its anti-apoptotic activity. This inhibition of Survivin phosphorylation occurred due to the direct suppression of Aurora-B by Go6976 treatment. These data indicate that Go6976 may have a unique therapeutic potential for patients with FLT3-driven leukemias.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1806. doi:1538-7445.AM2012-1806
Abstract 2413: Human glypican-3 promotes hepatocellular carcinoma progression via induction of epithelial-mesenchymal transition
Authors: Ogunwobi Olorunseun O. , Trinh Thu L. , Liu Chen
Published: August 2012

Hepatocellular carcinoma (HCC) is the third most important cause of cancer mortality worldwide. Epithelial-mesenchymal transition (EMT) has been shown to be an important biological process involved in the development and progression of HCC. The factors driving this process, however, are still largely unclear. Glypican-3 (GPC3) is upregulated in HCC and this is associated with poor prognosis. The details of the biological role of GPC3 in HCC are, however, yet to be fully elucidated. We hypothesized that GPC3 may promote HCC progression by inducing EMT. In this study we have examined the effect of human GPC3 in the progression of HCC using a murine HCC cell line (1MEA) derived from Balb/c mouse. We observed that the 1MEA cell line does not express GPC3 and we stably transfected human GPC3 into this cell line. 1MEA cells stably transfected with human glypican-3 (1MEAhGPC3) were then transplanted subcutaneously into 8 Balb/c mice. In comparison, wild type 1MEA was also transplanted into 8 separate Balb/c mice. Further, we performed a comparative analysis of the gene expression of E-cadherin, vimentin, collagen I, fibronectin, twist, slug and snail in both wild type 1MEA and 1MEAhGPC3. 18 days post-transplantation, tumor volume was analyzed. 5 mice transplanted with wild type 1MEA had not yet developed tumors whereas 3 mice transplanted with 1MEAhGPC3 developed tumors that had grown past the allowed tumor volume for the protocol and were sacrificed. Analysis of the remaining mice (3 transplanted with 1MEA and 5 with 1MEAhGPC3 revealed that tumors from 1MEA were 8-fold more voluminous than tumors from 1MEAhGPC3. Comparative analysis of the gene expression profile of 1MEA versus 1MEAhGPC3 revealed that transfection with hGPC3 had resulted in increased expression of mesenchymal markers (collagen I by 27.5-fold, fibronectin by 18-fold, vimentin by 3-fold), loss of epithelial marker (E-cadherin loss by 99%) and increased expression of transcriptional regulators of EMT (twist by 11.7-fold, slug by 148-fold and snail by 8.7-fold).These data clearly show that hGPC3 promotes HCC progression by inducing EMT in HCC cells. To our knowledge, this is the first experimental evidence that hGPC3's role in HCC is associated with induction of EMT and it suggests that therapeutic targeting of GPC3 in human HCC may be an effective approach to the treatment of advanced HCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2413. doi:1538-7445.AM2012-2413
Abstract 2418: NADPH Oxidase 1 over-expression enhances invasion via matrix metalloproteinase-2 and epithelial-mesenchymal transition in melanoma cells
Authors: Liu Feng , Garcia Angela , Meyskens Frank
Published: August 2012

Nox1 is a member of the NADPH Oxidase family which has not been well characterized in the melanocytic cell lineage. Here we demonstrated that Nox1 and Nox4 were detected in melanocytic lineage, with only Nox1 detected in normal human melanocytes and Nox4 in a subset of metastatic melanoma cell lines. The protein level and enzymatic activity of Nox1 was elevated in all melanoma cells as compared to normal melanocytes. Over-expression of GFP-Nox1 protein in Wm3211 primary melanoma cells increased invasion rate by 4-6 fold as measured by Matrigel invasion assay, while knocking down or inhibiting Nox1 decreased invasion by approximately 40 to 60%. MMP-2 was increased by Nox1 over-expression at the mRNA, protein and activity levels, and decreased by Nox1 knockdown. MMP-2 promoter activity was also regulated by Nox1 knockdown. In addition, stable clones over-expressing Nox1 exhibited an epithelial-mesenchymal transition (EMT) as examined by cell morphology and EMT markers; knockdown or inhibiting Nox1 led to a reversal of EMT. Supplementing MMP-2 to culture media did not induce EMT, suggesting that EMT induction by Nox1 was not through MMP-2 up-regulation. In summary, Nox1 was over-expressed in all melanoma cell lines examined, and enhanced cell invasion by MMP-2 up-regulation and EMT induction.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2418. doi:1538-7445.AM2012-2418
Abstract 2419: Irradiation of α-ionizing radiation to in vivo glioma model induced metastasis via occurrence of epithelial-mesenchymal transition
Authors: Park Jong Kuk , Jang Su Jin , Kang Sung Wook , Kim Jae-Sung , Park Sunhoo , Hwang Sang-Gu , Kang Joo Hyun , Um Hong Duck
Published: August 2012

α-Ionizing radiation (IR) therapy is known to be a one of major therapeutic tools of cancer treatment but α-IR also could be an inducer of tumorigenesis and metastasis. Metastasis is major obstacle and required to overcome for cancer treatment. Therefore, the aim of this investigation was to study whether radiation could induce metastasis our in vivo mouse system, and molecular mechanism behind α-IR-induced metastasis. C6L transfectant cell line expressing firefly luciferase (fLuc) was treated with α-IR and then immunoblotting, zymography and invasion assay in vitro. We also used C6L transfectant cell line to construct xenograft in nude mice and then IR was irradiated to xenograft with α-IR. The irradiated xenograft-containing mice were analyzed with survival curve, measurement of tumor size, and bioluminescence imaging in vivo and ex vivo. We also analyzed metastatic foci in organ of mice with RT-PCR analysis, H & E staining and immunohistochemistry. α-IR treatment to C6L cell induced ocurrence of epithelial-mesenchymal transition (EMT) and increase of cell invasion. In irradiated xenograft-containing mice, sizes of tumor were decreased dramatically and survival rate also extended. Almost all non-irradiated xenograft-containing control mice were dead within 4 weeks. But we also observed luminescence signals in about 20.9% of α-IR treated mice. Intestine or lung of luminescence signal-detected mice contained several foci, which expressed fLuc gene and presented histological features as a cancer tissue as well as expressions of EMT markers. Taken together, these findings indicate that α-IR treatment could induce metastasis in vivo via EMT including increase of MMP activity. We also propose that construction of metastasis animal model by α-IR treatment could use drug development to block cancer metastasis and increase understandings of metastasis mechanism.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2419. doi:1538-7445.AM2012-2419
Abstract 2420: Loss of hCLCA4 promotes EMT in breast cancer
Authors: Yu Yang , Walia Vijay , Elble Randolph C.
Published: August 2012

The epithelial to mesenchymal transition (EMT) is a developmental program in which epithelial cells downregulate their cell-cell junctions, acquire spindle cell morphology and exhibit cellular motility. In breast cancer, EMT facilitates invasion of surrounding tissues and correlates closely with cancer metastasis and relapse. Previous studies in our lab showed that the candidate tumor suppressor hCLCA2 is expressed in differentiated, growth-arrested mammary epithelial cells but is downregulated during tumor progression and EMT. We further showed that hCLCA2 is a p53-inducible growth inhibitor whose loss indicates an increased risk of metastasis. Subsequent studies have revealed that another member of the CLCA gene family, hCLCA4, is also expressed in mammary epithelial levels, albeit at lower levels than hCLCA2. We find that hCLCA4 is also induced by stress and is similarly downregulated in breast cancer cell lines. Like hCLCA2, hCLCA4 is downregulated when EMT is induced by TGF beta, ectopic expression of mesenchymal transcription factors, or dilution of immortalized cells. Furthermore, knockdown of hCLCA4 in HMLE by shRNAs caused EMT. RNA and protein profiling for EMT markers showed that knockdown cells have lower expression epithelial marker E-cadherin and higher expression level of mesenchymal markers vimentin and fibronectin. Moreover, we found that hCLCA4 is co-regulated with hCLCA2 in HMLE cells. Knockdown of hCLCA2 led to down-regulation of hCLCA4 and vice versa. These findings suggest that hCLCA4 is, like hCLCA2, a marker of epithelial differentiation whose loss contributes to EMT and possibly metastasis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2420. doi:1538-7445.AM2012-2420
Abstract 2421: Iron deficiency suppresses EMT through down-regulation of N-cadherin in esophageal cancer.
Authors: Watanabe Shinichiro , Nishitani Seishi , Noma Kazuhiro , Ohara Toshiaki , Tomono Yasuko , Hashimoto Yuuri , Tazawa Hiroshi , Fujiwara Toshiyoshi
Published: August 2012

[Introduction] Epithelial-Mesenchymal-Transition(EMT) plays a great role in cancer metastasis and invasion. Mechanism of EMT still remains unclear, However E-cadherin and N-cadherin are known to have contribution to EMT. In particular, down-regulation of N-cadherin has been reported to reduce cancer metastasis and invasiveness. Iron metabolism and its relationship with cancer cell were studied for a long time. Iron overload is known to induce some kinds of cancer, and iron deficiency is also known to suppress the tumor growth in vivo study. But the correlation of iron metabolism and tumor progression isn't unclear. Recently, it was reported that iron metabolism was essential for EMT and iron deficiency down-regulated EMT differentiation. In this study, we hypothesized that iron deficiency down-regulated the expression of N-cadherin, and suppressed the ability of metastasis and invasiveness. Here we demonstrate our results. [Materials and Methods] In vitro study, TE-4 and TE-10 esophageal squamous cell carcinoma and OE19 esophageal adenocarcinoma cell line were used in this study. Cell viability was determined by XTT assay. Iron chelator deferasirox was used to simulate iron deficient condition. To determine that iron deficiency suppresses EMT and invasiveness of cancer cells, we investigated western blotting analysis, three-dimensional spheroid model analysis and migration assay. Moreover, to determine the expression N-cadherin in mRNA level, PCR assay was performed. In vivo study, iron depletion condition was made by iron deficient diet. Normal and iron deficient diet were fed for 3 weeks before implantation of TE-4 cells in athymic male nude mice(n=8/group). After implantation, each diet was fed continuously for a month. [Results] In vivo study, iron deficient diet suppressed the tumor growth(tumor volume: normal diet vs iron deficient diet = 30.7±1.1 mm3 vs 10.9±2.2 mm3). In vitro study, deferasirox reduces proliferation of all cell lines in a concentration manner. According to western blotting analysis, deferasirox induces reduction of N-cadherin protein in TE-4 and TE-10 cell lines. N-cadherin down-regulation declines invasiveness of TE-4 and TE-10 cell lines in the three-dimensional spheroid model analysis and migration assay. Real time PCR assay shows that down-regulation of N-cadherin is dependent on reduction of N-cadherin mRNA. [Conclusion] Iron controlled treatment can be a novel therapy for invasiveness of cancer under EMT.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2421. doi:1538-7445.AM2012-2421
Abstract 2425: Collapsin response mediator protein 4 promotes tumor cell invasion and lethality of pancreatic cancer
Authors: Hiroshima Yukihiko , Nakamura Fumio , Miyamoto Hiroshi , Mori Ryutaro , Taniguchi Koichi , Matsuyama Ryusei , Takeda Kazuhisa , Akiyama Hirotoshi , Tanaka Kuniya , Ichikawa Yasushi , Nagashima Yoji , Goshima Yoshio , Endo Itaru
Published: August 2012

BACKGROUND: Pancreatic cancer is an aggressive malignancy with one of the worst outcomes among all cancers. Recently, Collapsin Response Mediator Proteins (CRMPs) were associated with proliferation, apoptosis, differentiation and invasion in several cancers. However, CRMPs expression and their role in pancreatic cancer have not been investigated. This study aimed to clarify the clinical significance of CRMPs in pancreatic cancer. METHODS: Pancreatic adenocarcinoma samples were obtained from 53 patients. Crmp expression in these samples was examined using realtime RT-PCR. CRMP4 protein expression was examined immunohistochemically and compared with the clinicopathological features of the tumors. Furthermore, knock down of CRMP4 expression using siRNA was examined in pancreatic cancer cell lines to determine whether CRMP4 regulates cell proliferation and invasion in vitro. RESULTS: Of all the CRMPs, the crmp4 gene was determined to be the only one differentially expressed in pancreatic cancer tissues (p = 0.008). The expression of CRMP4 was detected immunohistochemically in 34 of the 53 (64.1%) pancreatic cancer samples, and CRMP4 staining was highly correlated with poor differentiation (p = 0.003) and liver metastasis (p = 0.021). Multivariate analyses suggested that venous invasion and CRMP4 overexpression were prognostic factors for survival. CRMP4 knockdown using siRNA reduced cellular invasion, but did not affect proliferation. CONCLUSIONS: Our results suggested that CRMP4 overexpression was a strong prognostic factor for increasing pancreatic cancer invasiveness, and can be a novel therapeutic target for pancreatic cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2425. doi:1538-7445.AM2012-2425