bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2516: SL01, an oral gemcitabine prodrug as a potent anticancer drug
Authors: Li Wenbao , Yuan Hua , Qu Xianjun
Published: May 2015

SL01,dodecyl-3-((1-((2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)-tetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)carbamoyl)pyrazine-2-carboxylate, is a gemcitabine prodrug synthesized by introducing the moiety of 3-(dodecyloxycarbonyl)pyrazine-2-carbonyl at the N4-position on the cytidine ring of gemcitabine. Our goal is to investigate the efficacy and the safety of this oral prodrug as a potent oral anticancer agent, which may supplant the use of gemcitabine in clinic. The evaluation was accomplished by determining its inhibition of the growth of several tumors with gemcitabine administrated by injection as a control, and also by exploring its pharmacokinetics and its safety. In vivo, SL01 could effectively inhibit the growth of NCI-H460 human non-small cell lung cancer, MCF-7 human breast cancer, BxPC-3 human pancreatic cancer, ES-2 human bladder cancer, HepG2 human hepatocellular carcinoma cells and HCT-116 human colon adenocarcinoma cells in xenografts nude mice without loss their body weight significantly. After three weeks oral administration at the dosage of 30 umol/kg, the inhibition rates were 58.0%, 66.5%, 41.5%, 77.1%, 41.5% and 63.1%, respectively. Molecular analysis indicated that the high efficacy of SL01 was associated with its ability in inducing apoptosis, as evidenced by increase of TUNEL staining cells, activation of caspase-9, caspase-3 and cleaved poly ADP-ribose polymerase in tumor tissues. Pharmacokinetics studies indicated that SL01 exhibited a longer mean retention time and half-life than that of gemcitabine after oral administration in rat, further analysis showed that the bioavailability of SL01 was 22.1%, which indicating that the introduction of 3-(dodecyloxycarbonyl)pyrazine-2-carbonyl moiety to gemcitabine could improve its bioavailability and reduce its first-pass metabolism. The metabolism analysis suggested that SL01 could transform into gemcitabine by hydrolyzing and then releasing into blood to keep the relatively long steady-state concentrations. Toxicity studies indicated that the LD50 was >1.4 g/kg for its acute toxicity, and the tolerated dosage was 45 mg/kg for 4 weeks evaluation in rat. Based on these results, SL01 is proposed as a potent oral anticancer agent, which may supplant the use of gemcitabine in clinic. Acknowledgment: This project was supported by the Ministry of Science and Technology of China (2010ZX09401-302-2-07and 2011ZX09102-001-03).

Citation Format: Wenbao Li, Hua Yuan, Xianjun Qu. SL01, an oral gemcitabine prodrug as a potent anticancer drug. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2516. doi:10.1158/1538-7445.AM2014-2516
Abstract 2530: Optimization of 5,6-fused pyrimidine-based kinase inhibitors by computer-aided drug design for the treatment of AML
Authors: Hsu Yung Chang , Shiao Hui-Yi , Ke Yi-Yu , Hsu John T. A. , Lin Wen-Hsing , Chen Chun-Hwa , Hsieh Hsing-Pang
Published: May 2015

FMS-like receptor tyrosine kinase-3 (FLT3), a member of the class III tyrosine kinase receptor family, is expressed by immature hematopoietic cells, and highly associates with acute myeloid leukemia (AML). While acquired resistance to selective FLT3 inhibitors, the treatment became inefficient. Many literatures suggested that dual FLT3-Aurora inhibition may overcome selective FLT3 inhibitor resistance, in part due to inhibition of Aurora kinase, and may benefit patients with FLT3-mutated AML. Thus, utilizing dual FLT3-Aurora inhibitors may be the viable therapy for the treatment of AML.

Recently, our group reported a novel AURKA inhibitor by using high-throughput parallel synthesis and structure-based drug design (SBDD). Based on this, we wanted to modify the selective AURKA inhibitor to maintain its Aurora A inhibition as well as improve its FLT3 activity to develop dual FLT3-Aurora kinase inhibitors. Through rapid construction of the platform of FLT3 virtual screening, the lead was quickly and efficiently identified. Proceeding investigation of structure-activity relationship could lead to 6-phenyl pyrrolopyrimidine analogue, showing FLT3 inhibition with an IC50 of 101 nM as well as Aurora kinase A inhibition with an IC50 of 33 nM. To further increase the level of inhibition of FLT3, binding energy calculations and docking study of FLT3 model were introduced to select terminal groups of the side chain with high priority. The selected compounds were synthesized and evaluated the bioactivity. The dramatic antiproliferation of the dual FLT3-AURKA inhibitor against MOLM-13 and MV4-11 (FLT3-ITD (+) human AML cell lines) was observed, and its biologic mechanism of inhibition of FLT3 and Aurora kinase A was proved the by significant evidence of biomarker. For this finding, this novel inhibitor holds potential for further development of preclinical investigations as an anti-AML drug candidate.

Citation Format: Yung Chang Hsu, Hui-Yi Shiao, Yi-Yu Ke, John T. A. Hsu, Wen-Hsing Lin, Chun-Hwa Chen, Hsing-Pang Hsieh. Optimization of 5,6-fused pyrimidine-based kinase inhibitors by computer-aided drug design for the treatment of AML. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2530. doi:10.1158/1538-7445.AM2014-2530
Abstract 2554: DDX3X-specific effector T cells in small cell lung cancer patients reflect disease stage
Authors: Igarashi Natsue , Kagamu Hiroshi , Nozaki Koichiro , Shoji Satoshi , Okajima Masaaki , Miura Satoru , Watanabe Satoshi , Yoshizawa Hirohisa , Narita Ichiei
Published: May 2015

[Background] Small cell lung cancer (SCLC) possesses high tendency to disseminate. However, SCLC patients with paraneoplastic syndrome mediated by immunity against onconeural antigens remain in limited-stage disease (LD) without distant metastases. We previously demonstrated that effector-dominant CD4+ T cell balance was observed only in LD-SCLC but not in extended-stage disease (ED)-SCLC. CD4+ T cell balance in peripheral blood of SCLC patients reflected disease stage and recurrence.

Accumulating evidence suggests that most solid malignancies consist of heterogeneous tumor cells and that a relatively small subpopulation, which shares biological features with stem cells, is responsible for distant metastases and recurrence. We previously reported that DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide 3, X-linked (DDX3X) is an immunogenic protein preferentially expressed in CD133+ murine melanoma cells exhibiting biological CSC features and that DDX3X-primed CD4+ T cells mediated potent antitumor therapeutic efficacy. We found that DDX3X is strongly expressed in small cell lung cancer (SCLC).

[Objectives] In the current study, we examine that DDX3X-specific effector T cells exist in SCLC patients.

[Patients and Methods] The present study comprised 20 consecutive SCLC patients, 5 long-term survivors, and 10 healthy volunteers from a single institution (Niigata University Medical and Dental Hospital). Specimens were collected after obtaining written informed consent approved by the Niigata University Ethical Committee.

[Results] Six of 12 LD-SCLC patients possessed DDX3X-responsive effector T cells. CD62Llow CD4+ effector T cells obtained from peripheral blood of 5 LD-SCLC secreted significantly more IFNγ upon DDX3X antigen stimulation in the presence of CD11c+ autologous dendritic cells. CD62Llow CD8+ effector T cells from one LD-SCLC patient responded to DDX3X. In contrast, effector T cells obtained from ED-SCLC patients or healthy volunteers never responded to DDX3X.

[Conclusion] DDX3X is likely one of major antigens that is expressed in SCLC and is recognized by T cell immune system. Antitumor immunotherapy to induce effector-dominant CD4+ T-cell immunity on DDX3X antigen may be a promising therapy to eliminate SCLC cells that mediate distant metastases.

Citation Format: Natsue Igarashi, Hiroshi Kagamu, Koichiro Nozaki, Satoshi Shoji, Masaaki Okajima, Satoru Miura, Satoshi Watanabe, Hirohisa Yoshizawa, Ichiei Narita. DDX3X-specific effector T cells in small cell lung cancer patients reflect disease stage. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2554. doi:10.1158/1538-7445.AM2014-2554
Abstract 2573: The immunocytokine NHS-IL12 as a potential cancer therapeutic
Authors: Fallon Jonathan K. , Tighe Robert , Lyon Amanda J. , Sabzevari Helen , Schlom Jeffrey , Greiner John W.
Published: May 2015

Targeted delivery of IL-12 might make this cytokine a safer, more effective cancer therapeutic. Here we evaluated whether fusing IL-12 to a tumor necrosis-targeting human IgG1 (NHS76) could improve its anti-tumor efficacy in vivo. In vivo imaging and immunohistochemistry staining were used to assess the ability of this novel immunocytokine (NHS-IL12) to target murine tumors. The anti-tumor efficacy of NHS-IL12 was examined at various doses and treatment schedules. Immune cell subset-depleting antibodies, flow cytometric methods, and in vitro cytotoxicity and ELISA assays were used to investigate the mechanisms of action of NHS-IL12. This immunocytokine was investigated in several models and in all cases NHS-IL12 outperformed recombinant IL-12 in anti-tumor activity. Depletion of immune cell subsets revealed that the anti-tumor activity depends primarily on CD8+ cells; mice that had complete tumor regression following NHS-IL12 treatment displayed durable anti-tumor memory responses. Combining NHS-IL12 treatment with a cancer vaccine, radiation, or chemotherapy resulted in greater anti-tumor effects than each individual therapy alone. These preclinical findings provide a rationale for the clinical testing of this immunocytokine, both as a single agent and in combination with vaccines, radiation, and chemotherapy. NHS-IL12 is currently being evaluated in a Phase 1 clinical trial (NCT01417546).

Citation Format: Jonathan K. Fallon, Robert Tighe, Amanda J. Lyon, Helen Sabzevari, Jeffrey Schlom, John W. Greiner. The immunocytokine NHS-IL12 as a potential cancer therapeutic. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2573. doi:10.1158/1538-7445.AM2014-2573
Abstract 2584: Complete regression and protective immune responses obtained in B16 melanomas after treatment with LTX-315 (Oncopore®)
Authors: Camilio Ketil André , Berge Gerd , Ravuri Chandra Sekhar , Rekdal Øystein , Sveinbjørnsson Baldur
Published: May 2015

Background: Malignant melanoma is the most aggressive and deadliest form of skin cancer, with a high mortality rate among late-stage melanoma patients. This calls for new and improved therapies within the field. Cationic antimicrobial peptides (CAPs) are naturally occurring molecules found in most species, in which they play a significant role in the first line of defense against pathogens, and several CAPs have shown promising anticancer activity. Structure-activity relationship studies on the CAP bovine lactoferricin has resulted in the design of a short chemically modified lytic anticancer peptide, LTX-315, which has potential as a new anticancer drug. Previous animal studies conducted with LTX-315 have demonstrated that intratumoral (i.t.) treatment of syngeneic murine A20 B-cell lymphomas and CT26WT colon carcinomas resulted in complete tumor regression. In the present study, we investigated the antitumor effects of LTX-315 against highly aggressive B16 melanomas in syngeneic mice. Methods: The cytotoxicity of LTX-315 was tested against a series of cell lines in vitro using the MTT assay. B16F1 melanoma cells (5 x 104) were intradermally inoculated into the abdomen of syngeneic C57BL/6N mice and established tumors treated i.t. with LTX-315. Tumor tissue and blood samples were taken for analysis to elaborate the mechanism of action of LTX-315 and for potential immune modulatory effects following i.t. treatment. Blood plasma was analyzed for IL 1β and IL 6 content and tumor tissue was analyzed for infiltrating immune cells and expression of pro-inflammatory cytokines using immunohistochemistry and real time qPCR, respectively. Results: LTX-315 rapidly kills B16 melanoma cells and induces the release of Danger-Associated Molecular Pattern molecules such as the High Mobility Group Box-1 protein in vitro. Intratumoral (i.t.) administration of LTX-315 resulted in tumor necrosis and the infiltration of immune cells into the tumor parenchyma followed by a complete regression of the tumor in the majority of the animals. Animals cured by LTX-315 treatment were protected against a re-challenge with live B16 tumor cells. Conclusions: Our results demonstrate that LTX-315 i.t. treatments with LTX-315 induces complete regression of solid B16F1 melanoma tumors, thus leading to local tumor control, followed by protective immune responses. Thus, LTX-315 has potential as a novel immunotherapeutic agent.

Citation Format: Ketil André Camilio, Gerd Berge, Chandra Sekhar Ravuri, Øystein Rekdal, Baldur Sveinbjørnsson. Complete regression and protective immune responses obtained in B16 melanomas after treatment with LTX-315 (Oncopore®). [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2584. doi:10.1158/1538-7445.AM2014-2584
Abstract 2593: Multitargeted receptor tyrosine kinase inhibitors synergize with folate-hapten mediated immunotherapy in folate receptor positive cancers
Authors: Bandara Nimalka A. , Low Philip S.
Published: May 2015

The folate receptor (FR) which binds the vitamin folic acid with high affinity is overexpressed by a variety of human cancers including ovarian, endometrial, renal, and lung cancers. Consequently, the receptor has been exploited for the delivery of imaging agents and highly potent drugs exclusively to FR expressing tumors while avoiding uptake by the surrounding healthy tissue. This targeting mechanism can also be utilized for immunotherapy by vaccinating mice against a particular hapten in order to induce the production of anti-hapten antibodies and subsequently treating tumor bearing mice with a folate-hapten conjugate. The resulting reduction in tumor burden is achieved by antibody-dependent cellular cytotoxicity (ADCC). However, hapten mediated immunotherapy alone has been ineffective at controlling tumor progression over longer periods of time and requires supplementation with immunomodulatory cytokines for optimum therapeutic results. Several multitargeted receptor tyrosine kinase (RTK) inhibitors have recently shown promising synergy in a variety of different cancers when combined with other immunotherapies. Therefore, we examined the effect of combining the RTK inhibitors sunitinib and axitinib with folate-fluorescein therapy in tumor bearing mice that had previously been immunized against fluorescein. We report here that sunitinib synergizes with folate-fluorescein in both the L1210A lymphoma and the M109 lung cancer models resulting in slowed tumor progression, inhibition of tumor vasculature, and increased levels of circulating CD4+ and CD8+ T cells. The combination of axitinib with folate-fluorescein was only evaluated in mice bearing L1210A tumors and these mice have also demonstrated lower average tumor volumes than mice treated with either drug alone or not treated at all. These results suggest that RTK inhibitors play an important role in modulating the anti-tumor immunity of tumor bearing mice and could be a potent addition to ADCC mediated therapeutic approaches for cancers.

Citation Format: Nimalka A. Bandara, Philip S. Low. Multitargeted receptor tyrosine kinase inhibitors synergize with folate-hapten mediated immunotherapy in folate receptor positive cancers. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2593. doi:10.1158/1538-7445.AM2014-2593
Abstract 2596: Suppression of lung cancer malignancy by peptides targeting MARCKS phosphorylation site domain: A comparative study
Authors: Chen Ching-Hsien , Statt Sarah , Thai Phillip , Lee Jasmine G. , Adler Kenneth B. , Yang Pan-Chyr , Wu Reen
Published: May 2015

We have recently reported elevated MARCKS phosphorylation (p-MARCKS) in lung cancer tissues but not in their adjacent normal non-cancer tissue sections. We have extended this study to a tumor tissue array from a cohort of 110 human patients with lung cancer. Immunohistochemistry results have shown a significant association of elevated p-MARCKS with advanced-stage, lymph node metastasis and malignant phenotypes of lung cancer, confirming the importance of p-MARCKS in lung cancer progression. To see if this newly discover biomarker can be a target for therapeutic treatment for the suppression of lung cancer malignancy, we have employed two peptides for this potential; one is the MANS peptide developed by our collaborator many years ago to target myristoylated N-terminal amino acid region for the control of airway mucus granule secretion, the other peptide is MPS that directly targets the MARCKS phosphorylation site domain. For MANS peptide, our recent publication has shown it is effective in blocking lung cancer cell migration and invasiveness in vitro as well as in vivo metastasis. A similar observation was seen for MPS peptide treatment. However, in contrast to MANS peptide, MPS treatment can directly block PIP3-dependent AKT signaling and repress lung tumor growth in vivo in addition to the suppression of metastasis, while MANS can only reduce cancer metastasis. These results support the initial notion on the involvement of p-MARCKS as a biomarker in lung cancer and the potential to target the phosphorylation site domain for controlling cancer progression. Our data have also suggested a direct targeting MARCKS phosphorylation site domain by MPS peptide is more effective in the suppression of lung cancer malignancy.

Citation Format: Ching-Hsien Chen, Sarah Statt, Phillip Thai, Jasmine G. Lee, Kenneth B. Adler, Pan-Chyr Yang, Reen Wu. Suppression of lung cancer malignancy by peptides targeting MARCKS phosphorylation site domain: A comparative study. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2596. doi:10.1158/1538-7445.AM2014-2596
Abstract 2608: Dissecting Myc inhibition as a cancer therapy
Authors: Salisbury Nicholas J. H. , Wilson Catherine H. , Lu Dan , Littlewood Trevor D. , Evan Gerard I.
Published: May 2015

The MYC family of basic helix-loop-helix leucine zipper (bHLH-LZ) transcription factors are critical for cell growth, proliferation and apoptosis. c-Myc is deregulated in most, if not all, cancers making it an attractive target for cancer therapy. Previously we have shown that Myc inhibition by a mutant c-Myc derivative, Omomyc, leads to tumor regression in several cancer models, notably in murine KRasG12D-driven lung adenocarcinomas. The mechanism by which Omomyc expression leads to tumor regression has not been fully elucidated, nor do we know the biochemical properties of a small molecule that could replicate Omomyc's activity. Therefore, we designed two novel Omomyc mutants (ΔMyc and MMO) that are hypothesised not to interact with the full Omomyc interactome but are still able to inhibit Myc transactivation. We subsequently created human lung adenocarcinoma cell lines (A549s) and transgenic mice which inducibly express Omomyc, ΔMyc and MMO. Unexpectedly Omomyc and ΔMyc inhibited proliferation in vitro but not MMO. In vivo, expression of the mutants has been detected and we are currently assessing their effect on KRasG12D-driven adenocarcinomas.

Citation Format: Nicholas J. H. Salisbury, Catherine H. Wilson, Dan Lu, Trevor D. Littlewood, Gerard I. Evan. Dissecting Myc inhibition as a cancer therapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2608. doi:10.1158/1538-7445.AM2014-2608
Abstract 2609: The role of focal adhesion kinase (FAK), PI3K/AKT and p53/mouse double minute 2 homologue (MDM2) complex in the pathobiology of Merlin-deficient tumors
Authors: Hanemann Clemens O. , Ammoun Sylwia , Schmid Marei C. , Zhou Lu
Published: May 2015

Deficiency of the tumor suppressor merlin leads to the development of schwannomas, meningiomas and ependymomas occurring spontaneously or as a part of a hereditary disease Neurofibromatosis type 2 (NF2). Another tumor suppressor/transcription factor p53 regulates proliferation, survival and differentiation. P53 deficiency/inactivation correlates with tumor development and can be negatively regulated by FAK, PI3K/AKT and MDM2 and possibly positively by merlin and JNK in different models. The function of p53 in merlin deficient tumors is not known. Using our in vitro model of primary human schwannoma cells we demonstrated that FAK is overexpressed/activated and localises to the nucleus in schwannoma cells leading to increased proliferation. AKT is strongly activated via PDGF- and IGF-I- receptors increasing survival. Here we investigated p53 regulation and its role in proliferation and survival of human primary schwannoma cells. p53 is downregulated and MDM2 upregulated in human schwannoma cells leading to increased proliferation and survival. p53 is regulated by merlin involving JNK FAK, AKT and MDM2. Thus merlin potentiates p53 activity and when using Nutlin-3 this increases p53 stability and total levels of p53 in schwannoma cells decreasing proliferation and survival. These findings are important to dissect mechanisms responsible for development of merlin-deficient tumors and identification of therapeutic targets.

Citation Format: Clemens O. Hanemann, Sylwia Ammoun, Marei C. Schmid, Lu Zhou. The role of focal adhesion kinase (FAK), PI3K/AKT and p53/mouse double minute 2 homologue (MDM2) complex in the pathobiology of Merlin-deficient tumors. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2609. doi:10.1158/1538-7445.AM2014-2609
Abstract 2611: Function blocking ERBB3 antibody inhibits adaptive response to RAF inhibitor
Authors: Kugel Curtis Harrison , Hartsough Edward J. , Davies Michael A. , Aplin Andrew E.
Published: May 2015

ERBB3/HER3 is a member of the EGFR family of receptor tyrosine kinase. Our lab recently showed that treating melanoma cells with the FDAapproved small molecule BRAF inhibitor vemurafenib leads to an increase in ERBB3 signaling when grown in the presence of its ligand neuregulin (NRG1). Here, we show that targeting ERBB3 with a monoclonal antibody inhibits its activation and downstream signaling in the presence of vemurafenib and NRG1. This inhibition of ERBB3 led to a decrease in proliferation and increase in apoptosis in vitro as measured by EdU and AnnexinV/PI staining. Targeting ERBB3 in combination with mutant BRAF in vivo resulted in a delay of tumor re-growth in immune deficient mice compared to targeting mutant BRAF alone. Taken together, these results indicate that targeting ERBB3 in combination with mutant BRAF in melanoma may help enhance the initial effects of vemurafenib in the clinic.

Citation Format: Curtis Harrison Kugel, Edward J. Hartsough, Michael A. Davies, Andrew E. Aplin. Function blocking ERBB3 antibody inhibits adaptive response to RAF inhibitor. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2611. doi:10.1158/1538-7445.AM2014-2611
Abstract 2627: Use of xenografts for preclinical drug testing
Authors: Lange Mark J. De , Némati Fariba , Versluis Mieke , Jager Martine J. , Luyten Gre P.M. , Decaudin Didier , Velden Pieter A. van der
Published: May 2015

Purpose: Uveal melanoma (UM) originates from melanocytes just like cutaneous melanoma (CM) and similar to CM, the MAPK pathway is involved in the development of UM. However, in vitro we showed an inverse correlation between MAPK and c-Met activation with increasing metastatic potential of UM. In this study, we aimed to target both Src and c-Met with respectively Dasatinib and Crizotinib in an experimental xenograft model.

Methods: We used tissue derived from four different human primary tumors which were subcutaneously transplanted in mice. We used the c-Met inhibitor Crizotinib to inhibit c-Met function and Dasatinib to inhibit MAPK via Src. We monitored pharmacodynamics and analysed treatment efficacy with kinase assays.

Results: Growth inhibition was seen after treatment with Dasatinib in all four tumors whereas Crizotinib effects remained limited. Dasatinib inhibited kinase activity in the xenografts and pre-treatment kinase activity of the xenograft turned out to be predictive for treatment efficacy.

Conclusion: Activated Src and c-Met are in vitro susceptible to Dasatinib and Crizotinib, respectively. In vivo Dasatinib showed an effect on both tumor growth and MAPK activity while Crizotinib effects remained limited to c-Met inhibition. Combined, these results support our in vitro pathway analysis and warrant for clinical trials with Dasatinib in UM and further in vivo analysis with Crizotinib.

Citation Format: Mark J. De Lange, Fariba Némati, Mieke Versluis, Martine J. Jager, Gre P.M. Luyten, Didier Decaudin, Pieter A. van der Velden. Use of xenografts for preclinical drug testing. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2627. doi:10.1158/1538-7445.AM2014-2627
Abstract 2629: Nintedanib (BIBF-1120) inhibits hepatocellular carcinoma growth independent of angiokinase activity
Authors: Tai Wei-Tien , Shiau Chung-Wai , Li Yong-Shi , Chang Chun-Wei , Huang Jui-Wen , Hsueh Ting-Ting , Chen Kuen-Feng
Published: May 2015

Background & Aims: Nintedanib, a triple angiokinase inhibitor, is currently being evaluated against advanced HCC in phase I/II clinical trials. Here, we report the underlying molecular mechanism by which nintedanib (BIBF-1120) induces an anti-HCC effect.

Methods: HCC cell lines were treated with nintedanib or its derivative (ΔN) and apoptosis, signal transduction, and phosphatase activity were analyzed. Purified SHP-1 proteins or HCC cells expressing deletion N-SH2 domain or D61A point mutants were used to investigate the potential effect of nintedanib on SHP-1. In vivo efficacy was determined in nude mice with HCC subcutaneous xenografts (n ≥ 8 mice).

Results: Nintedanib induced anti-proliferation in HCC cell lines by targeting STAT3. Ectopic STAT3 abolished nintedanib-mediated apoptosis in HCC cells. Nintedanib further activated SHP-1 in purified SHP-1 proteins suggesting that nintedanib directly affects SHP-1 for STAT3 inhibition. HCC cells or recombinant SHP-1 proteins expressing deletion of N-SH2 domain or D61A mutants restored the activity of nintedanib suggesting that the auto-inhibition structure of SHP-1 was relieved by nintedanib. To further elucidate whether the effect of nintedanib on SHP-1 is dependent on its angiokinase inhibition activity, we developed a novel kinase-independent derivative of nintedanib, ΔN. Although ΔN only retained the backbone of nintedanib without kinase activity, ΔN still induced substantial anti-HCC activity in vitro and in vivo by targeting STAT3.

Conclusions: Nintedanib induced significant anti-HCC activity independent of angiokinase inhibition activity in a preclinical HCC model by relieving autoinhibition of SHP-1. Our findings provide new mechanistic insight into the inhibition of HCC growth by nintedanib.

Citation Format: Wei-Tien Tai, Chung-Wai Shiau, Yong-Shi Li, Chun-Wei Chang, Jui-Wen Huang, Ting-Ting Hsueh, Kuen-Feng Chen. Nintedanib (BIBF-1120) inhibits hepatocellular carcinoma growth independent of angiokinase activity. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2629. doi:10.1158/1538-7445.AM2014-2629
Abstract 2636: Connexin 43 suppresses tumor angiogenesis by downregulation of vascular endothelial growth factor
Authors: Chen Man-Chin , Lee Che-Hsin
Published: May 2015

Gap junction intercellular communication (GJIC), which is performed by connexins, is crucial in the regulation of cellular functions. Connexin 43 (Cx43) is a general isoform expressed in most epithelial tissues especially in astrocytes. There were some studies indicated that Cx43 played a tumor suppressor which has been found in human tumors compared with adjacent normal tissues. Otherwise, there were some evidences showed that Cx43 overexpression affected metastasis. However, Cx43 plays a controversial role in tumor progression. Hypoxia, a hallmark of many solid tumors, is associated with angiogenesis and tumor progression. It activates a signal cascade that culminates in the stabilization of hypoxia-inducible factor 1 (HIF-1) transcription factor and activation of genes that possess hypoxia response elements (HRE). Herein, the loss of Cx43 was shown to stabilize HIF-1α, which is overexpressed in a majority of cancers and its overexpression correlates with poor prognosis and treatment failure. Meanwhile, we examined HIF-1α expression and activities in cancer cells with various Cx43 statuses. Loss of Cx43 expression in cancer cells resulted in increased HIF-1-dependent transcriptional activity and protein expression. In this study, we also found that Cx43 down-regulated VEGF and inhibited endothelial cell proliferation. Further work is warranted to understand the molecular mechanism leading to Cx43 effects by antiangiogenic signal pathway.

Citation Format: Man-Chin Chen, Che-Hsin Lee. Connexin 43 suppresses tumor angiogenesis by downregulation of vascular endothelial growth factor. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2636. doi:10.1158/1538-7445.AM2014-2636
Abstract 2667: RG7287, a novel humanized anti-CDCP1 antibody with superior preclinical in vivo efficacy in combination with Paclitaxel
Authors: Kollmorgen Gwendlyn , Lifke Alexander , Nopora Adam , Bauss Frieder , Niederfellner Gerhard , Bossenmaier Birgit
Published: May 2015

CUB domain-containing protein 1 (CDCP1) is a transmembrane glycoprotein and a substrate of Src family kinase (SFK). It has been shown to be upregulated and significantly contributing to a number of different cancers, including colon, lung, breast, kidney, and pancreas cancer. High expression of CDCP1 has been shown to correlate with poor prognosis. CDCP1 is involved in the regulation of anoikis resistance, cell migration and invasion. We have recently described RG7287, a humanized glycoengineered therapeutic anti-CDCP1 antibody with a dual mode of action. (1) downregulation of the receptor from the cell surface, and (2) engaging immune effector functions such as antibody-dependent cellular cytotoxicity (ADCC). Upon ligation of RG7287 to CDCP1 an initial phosphorylation of CDCP1 occurs, this transient activation results in the down-regulation of CDCP1. RG7287 prohibits the transformation potential of CDCP1 and Src in focus formation assays. In vivo, RG7287 increased the survival time of mice bearing tumors of MCF7 cells stably expressing CDCP1 compared to untreated CDCP1 overexpressing MCF7 tumors. Treatment of three different xenograft models with RG7287 inhibited tumor growth.

In this study we looked at the possibility of eradicating the tumors by combining RG7287 with paclitaxel. Combining the cytostatic with RG7287 increases tumor growth inhibition compared to RG7287 alone in one xenograft model. In another model the combination of RG7287 with paclitaxel leads to tumor stasis. These results indicate a potential combination therapy for RG7287. We also tested whether RG7287 inhibits metastasis in vivo. Using a metastasis model we could show convincing inhibition of metastasis.

Citation Format: Gwendlyn Kollmorgen, Alexander Lifke, Adam Nopora, Frieder Bauss, Gerhard Niederfellner, Birgit Bossenmaier. RG7287, a novel humanized anti-CDCP1 antibody with superior preclinical in vivo efficacy in combination with Paclitaxel. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2667. doi:10.1158/1538-7445.AM2014-2667
Abstract 2677: Lysyl Oxidase a therapeutic target in pancreatic cancer
Authors: Saturno Grazia , Lopes Filipa , Viros Amaya , Morton Jennifer , Sansom Owen , Springer Caroline , Marais Richard
Published: May 2015

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies, with a 5-year survival rate of only 6%. Despite advances in conventional therapies and targeted therapies, survival has not improved in the past three decades. We found that high expression of LOX (Lysyl Oxidase) is a poor prognostic marker in PDAC, suggesting a role in this disease. LOX is an amine oxidase that catalyzes the cross-linking of collagens and elastin in the extracellular matrix (ECM) thus regulating extracellular stiffness and contributing to desmoplasia (abundant fibrotic stroma). LOX inhibition has been associated with decreased phosphorylation of SRC (Proto-oncogene tyrosine-protein kinase Src) and FAK (Focal Adhesion Kinase) as a result of decreased interaction of integrins with the ECM. To further investigate the role of LOX in pancreatic cancer, we obtained PDAC cells derived from a transgenic mouse model that faithfully recapitulates the human disease. We found that LOX was upregulated in tumor cells from Pdx1-Cre KrasG12D/+ Trp53R172H/+ (KPC) mice compared to Pdx1-Cre KrasG12D/+ (KC) mice. Using a genetic approach we developed LOX-relevant models by silencing LOX in KPC cells via shRNA and overexpressing LOX in KC cells. We characterized these cells for their invasion ability and growth in a 3D environment. We then confirmed these in vitro results in allograft models in mice. Collectively, our results validate LOX as a therapeutic target in PDAC and suggest that its inhibition represents a new therapeutic strategy for PDAC patients.

Citation Format: Grazia Saturno, Filipa Lopes, Amaya Viros, Jennifer Morton, Owen Sansom, Caroline Springer, Richard Marais. Lysyl Oxidase a therapeutic target in pancreatic cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2677. doi:10.1158/1538-7445.AM2014-2677
Abstract 2683: Inhibition of BCAT1 suppresses the expression of pro-metastatic proteins and reduces cancer metastasis
Authors: Papathanassiu Adonia E. , Vu Hong A.
Published: May 2015

Recent evidence suggests that the cytosolic presence of branched chain amino acid aminotransferase (BCAT1) in adult tissues is associated with diverse pathological conditions including cancer and inflammatory diseases such as rheumatoid arthritis; in these diseases, BCAT1 appears to modulate the expression of many key proteins and to be a new druggable target. In cancer, BCAT1 physically associates with CD147 and regulates the levels and the cellular localization of the protein in a number of different cell lines in vitro and in vivo. Silencing of Bcat1 gene downregulates the total expression of CD147 in MDA-MB-435 cells, while inhibition of BCAT1 by sodium 4-methyl-5-oxo-hexanoate (ERG240) leads to suppression of the extracellular CD147 in the same cells. In turn, suppression of extracellular CD147 results in downregulation of CD147-associated proteins such as MCT4, matrix metalloproteinases (MMPs), cyclophilin A, and GAPDH. Furthermore, inhibition of BCAT1 is associated with prevention of cancer cell migration and reduction in secretion of metabolic by-products such as lactate. Surprisingly, treatment of MDA-MB-231 tumor-bearing animals with 2-deoxyglucose (2DG), a well-known inhibitor of glycolysis, leads to a decrease in the levels of c-MYC and BCAT1, an increase in the total expression of CD147, and the increased shedding of the extracellular CD147 into the circulation. This observation suggests that BCAT1-driven regulation of CD147 in cancer cells involves a complex mechanism. Inhibition of tumoral BCAT1 is expected to adversely affect the metastatic potential of various tumors not only through a reduction in cell motility but also through alterations in the tumor microenvironment, driven by inhibition of extracellular CD147.

Citation Format: Adonia E. Papathanassiu, Hong A. Vu. Inhibition of BCAT1 suppresses the expression of pro-metastatic proteins and reduces cancer metastasis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2683. doi:10.1158/1538-7445.AM2014-2683
Abstract 2690: Sorafenib/regorafenib and phosphatidyl inositol 3 kinase/thymoma viral proto-oncogene inhibition interact to kill tumor cells
Authors: Booth Larry A. , Cruickshanks Nichola A. , Sajithlal G B. , Hamed Hossein A. , Tavallai Seyedmehrad , Syed J , Grant Steven , Poklepovic Andrew , Dent Paul
Published: May 2015

The present study aimed to determine whether the multikinase inhibitors, sorafenib/regorafenib, worked in conjunction with PI3K/AKT inhibitors to enhance tumor cell death. It has been noted that combination of sorafenib/regorafenib with a PI3K inhibitor acetic acid (PX-866) resulted in tumor cell death, in a greater than additive fashion, in liver, colorectal, lung, breast, kidney and brain cancer cells. Similar data was obtained using AKT inhibitors perifosine and MK2206. Furthermore, even cells lacking PTEN remained as sensitive to this combinational approach as cells expressing PTEN. PX-866 treatment abolished AKT/GSK3 phosphorylation, with tumor cell death correlating with reduced activity of AKT and mTOR. Expression of activated AKT and to a lesser extent activated mTOR reduced drug combination lethality. Expression of B-cell lymphoma-extra large or dominant negative caspase 9, but not cellular FLICE-inhibitory protein short, protected cells from the drug combination. Treatment of cells with PX-866 increased protein levels of p62, LAMP2 and LC3/LC3II which correlated with the resulting increase in LC3-GFP vesicle numbers. Exposure of PX-866 treated cells to sorafenib reduced p62 and LAMP2 levels, decreased the ratio of LC3 to LC3II and reduced LC3-GFP vesicle levels. Knockdown of Beclin1 or ATG5 suppressed drug toxicity by ∼40%. In vivo, combination of either sorafenib and PX-866 or regorafenib and MK2206 cooperated to suppress the growth of established HuH7 and HCT116 tumors, respectively. Collectively our data demonstrates that the combination of sorafenib family kinase inhibitors with inhibitors of the PI3K/AKT pathway kills tumor cells in vitro and in vivo.

Citation Format: Larry A. Booth, Nichola A. Cruickshanks, G B. Sajithlal, Hossein A. Hamed, Seyedmehrad Tavallai, J Syed, Steven Grant, Andrew Poklepovic, Paul Dent. Sorafenib/regorafenib and phosphatidyl inositol 3 kinase/thymoma viral proto-oncogene inhibition interact to kill tumor cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2690. doi:10.1158/1538-7445.AM2014-2690
Abstract 270: The association between medical history and breast cancer risk: A nationwide population-based case-control study using claim data
Authors: Chuang Shu-Chun , Wu Guo-Jie , Lu Yen-Shen , Lin Ching-Hung , Hsiung Chao Agnes
Published: May 2015

Introduction: It is suggested that chronic disease may be associated with increased cancer risk due to common risk factors, treatments to the prior conditions, or other unknown mechanisms. We evaluated the associations between prior medical conditions and subsequent breast cancer risk using the claim data from the Taiwan National Health Insurance (NHI) program.

Material and Method: A matched case-control study was conducted using the NHI 1,000,000 random sample dataset. A list of medical conditions based on literature review was retrieved for each case and control. The odds ratios (OR) and 95% confidence intervals (CI) of the association between medical conditions and breast cancer risk were estimated using conditional logistic regression and adjusted for occupation and number of breast cancer screening. The associations were estimated for younger (<50 years old) and older subjects, separately.

Results: Prior breast diseases, hypertensive diseases, and endometriosis were associated with increased breast cancer risk in both age groups. Thyroid diseases were associated with breast cancer in the younger (OR=1.32, 95% CI=1.04-1.66) but not in the older group (OR=1.02, 95% CI=0.82-1.27). On the other hand, metabolic syndrome, particularly the disorders involved lipid metabolism, was associated with breast cancer only in the older group (OR=1.24, 95% CI=1.04-1.49).

Conclusion: Our analyses support results from previous studies that prior breast diseases and endometriosis were associated with subsequent breast cancer at all ages, thyroid diseases were associated with premenopausal, and metabolic syndrome were associated with postmenopausal breast cancer.

Note: This abstract was not presented at the meeting.

Citation Format: Shu-Chun Chuang, Guo-Jie Wu, Yen-Shen Lu, Ching-Hung Lin, Chao Agnes Hsiung. The association between medical history and breast cancer risk: A nationwide population-based case-control study using claim data. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 270. doi:10.1158/1538-7445.AM2014-270
Abstract 2708: Development of allosteric regulators of the dimerization domains of the plasminogen-related growth factor family for the treatment of pancreatic cancer
Authors: Church Kevin J. , Kawas Leen , Harding Joseph , Lang Malte , McMicheal Michelle
Published: May 2015

The plasminogen-related family of growth factors includes the hepatocyte growth factor (HGF) and macrophage stimulating protein (MSP), both of which normally mediate a variety of cellular behaviors including proliferation, survival, and migration. A major contributor to the formation and persistence of various cancers is the dysregulation of these growth factor systems. Overexpression of the HGF receptor, c-Met, is common in many cancers including pancreatic cancer, and is associated with poor prognosis. Dysregulation of this system often leads to tumorigenesis and induction of various malignant phenotypes including increased proliferation, survival, migration, and invasion. In order to activate c-Met, HGF must first undergo an activation step in which it forms an HGF homodimer. Small molecule antagonists with homology to a putative “hinge” region within the HGF sequence have been developed that bind to HGF and allosterically block dimerization, therefore inhibiting c-Met signaling. Norleual [Nle-Tyr-Ile-ψ-(CH2-NH2)3-4-His-Pro-Phe] has previously been shown to inhibit HGF dimerization and HGF-dependent c-Met activation. We have assessed the ability of Norleual to block HGF dimerization and activation of c-Met within pancreatic cancer cells. Similar to the HGF/c-Met system, over-activation of the closely related MSP/Ron (MSP receptor) growth factor system has been identified as a critical contributor to pancreatic cancer progression. Due to overall high sequence homology between HGF and MSP, including the putative “hinge” region, we have hypothesized that MSP must also dimerize in order to efficiently activate the Ron receptor, and that this activation step can be targeted by “hinge” region homologs. Our results indicate that MSP naturally forms dimers and incubation with Norleual and similar “hinge” region antagonists inhibits dimerization. Further, we have demonstrated that MSP dimerization may be biologically significant and inhibition of dimerization suppresses MSP-dependent cell signaling and cell behaviors. The ultimate goal of this work is to develop a dual inhibitor capable of suppressing both the HGF and MSP growth factor systems, and further assess the therapeutic potential of these inhibitors for treatment of pancreatic cancer.

Citation Format: Kevin J. Church, Leen Kawas, Joseph Harding, Malte Lang, Michelle McMicheal. Development of allosteric regulators of the dimerization domains of the plasminogen-related growth factor family for the treatment of pancreatic cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2708. doi:10.1158/1538-7445.AM2014-2708