bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5495: Lurbinectedin (PM01183) in vivo synergizes the antitumor activity of taxanes.
Authors: Guillén Maria José , Cataluña Oscar , Palomares Mandy , López Raquel , Núñez Práxedes , Cuevas Carmen , Avilés Pablo
Published: August 2014

Background: Lurbinectedin (PM01183), a new synthetic tetrahydroisoquinoline alkaloid, is a minor groove specific DNA binder that interacts directly with specific factors involved in DNA repair and transcription pathways. In living cells, PM01183-DNA adducts stall replication and transcription giving rise to double strand breaks, inducing accumulation of cells in the S-phase of the cell cycle and triggering apoptosis. In both the ongoing phase II trials (pancreas and platinum-resistant ovarian cancer) the first stage was concluded with positive results in clinical activity, ensuring the continuity of the trials. Also, phase I trials in combination with doxorubicin and gemcitabine as well as in advanced acute leukemia are ongoing. In vivo studies aimed at defining the synergism of PM01183 combined with taxanes (taxol and docetaxel) are presented here.

Material and Methods: Athymic mice were implanted with HGC-27 (gastric), A2780 (ovarian), H460 (NSCLC), 22RV1 (prostate) or MDA-MB-231 (breast) cancer cells. Mice bearing tumors (ca. 150 mm3) were allocated (N=6-8/group) to experimental groups, namely: placebo; PM01183 at MTD (0.180 mg/kg [0.54 mg/m2]), 0.75MTD, 0.5MTD and 0.25MTD; taxol (H460, A2780 or HGC-27) or docetaxel (22RV1 or MDA-MB-231) at their MTD, 0.75MTD, 0.5MTD and 0.25MTD; and, PM01183 combined either with taxol or docetaxel at (1+1), (0.75+0.75), (0.50+0.50) and (0.25+0.25) MTDs. All treatments were given intravenously once per week during the placebo-treated survival time. The combination index (CI) was determined by the CI-isobol method. When needed, statistical differences for the antitumor effect recorded between groups were determined using two-tailed Mann-Whitney U test.

Results: In all the experiments, the combination of PM01183 and taxol or docetaxel resulted in more antitumor effect (P<0.05) than that obtained with the more active single agent at the highest dose (MTD level). Analysis of the results by the CI-isobol method indicated synergism (CI<1.0) after the treatment with PM01183 plus taxol of mice bearing HGC-27, A2780, H460 xenografted tumors. Also, in 22RV1 or MDA-MB-231 tumor bearing animals, the treatment of PM01183 plus docetaxel resulted in an additive or synergistic effect.Conclusion: An improved (additive or synergism) in vivo antitumor activity was recorded after the treatment with PM01183 plus taxanes (taxol or docetaxel) of mice bearing gastric, ovarian, NSCLC, breast or prostate xenografted tumors.

Citation Format: Maria José Guillén, Oscar Cataluña, Mandy Palomares, Raquel López, Práxedes Núñez, Carmen Cuevas, Pablo Avilés. Lurbinectedin (PM01183) in vivo synergizes the antitumor activity of taxanes. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5495. doi:10.1158/1538-7445.AM2013-5495
Abstract 5507: Mechanistic study of a new 4-(3, 5-dimethoxyphenyl)-N-(7-fluoro-3-methoxyquinoxalin-2-yl)piperazine-1-carboxamide compound (RX-5902).
Authors: Lee Young Bok , Kim Deog Joong , Kost Gina Chun , Frank Julie , Ahn Chang-Ho , Liu Zhi-Ren
Published: August 2014

In our previous study, we showed that a novel compound 1-(3,5-dimethoxyphenyl)-4-[(6-fluoro-2-methoxyquinoxalin-3-yl)aminocarbonyl] piperazine induces apoptosis of cancer cells by downregulation of Bcl-2 protein levels and causing G2/M cell cycle arrest. We observed also a synergistic effect in growth inhibition when combined with known anticancer agents in cancer cells and potent anti-growth activity in drug-resistant cancer cells as well as anti-proliferation of cancer cells at IC50 values of low nanomolar concentrations. Oral administration of the compound led to inhibition of tumor growth and enhanced survival in several tumor xenograft models. Our recent studies indicated that the compound interacts with p68 RNA helicase (also known as DDX5), a member of the DEAD box family of RNA helicases. p68 has been demonstrated to play a vital role in cell proliferation and tumor/cancer progression. Our studies showed that the compound did not exert its anti-cancer effects by inhibition of RNA unwinding by p68. Instead, the compound completely abrogated the β-catenin stimulated ATPase activity of p68 with an IC50 of 61 nM. More extensive studies are ongoing to validate and further characterize our interesting discovery.

Citation Format: Young Bok Lee, Deog Joong Kim, Gina Chun Kost, Julie Frank, Chang-Ho Ahn, Zhi-Ren Liu. Mechanistic study of a new 4-(3, 5-dimethoxyphenyl)-N-(7-fluoro-3-methoxyquinoxalin-2-yl)piperazine-1-carboxamide compound (RX-5902). [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5507. doi:10.1158/1538-7445.AM2013-5507
Abstract 5528: Identification of compounds modifying radiation-therapy using a 3D-microtissue technology.
Authors: Anastasov Natasa , Höfig Ines , Lichtenberg Jan , Stroebel Simon , Salomon Michael , Thirion Christian , Kelm Jens , Atkinson Michael J.
Published: August 2014

The two key issues in chemo- and radiation therapy is the development of tumor resistance as well as toxic effects on normal tissue. In this sense new strategies are required to increase efficacy of radiation to improve the therapeutic impact and reduce toxicological side effects. The performance of 3D cell culture systems over classical 2D culture systems has been shown to provide a closer representation of tissue-level biology. This has led to the rapid adoption of 3D systems for both drug discovery and toxicology. InSphero has developed a highly reproducible hanging drop technology able to generate monotypic cell spheroids (microtissues) in a 96-well format. The innovative 3D-microtissue plate technology has been adapted for analysis of the cellular response of radioresistant T47D breast cancer cells to combined radio-chemotherapy (RCTx). We have validated the model by comparing the treatment of microtissues with 10 different chemotherapeutic compounds, each tested alone and in combination with an acute 2Gy radiation exposure. The T47D cells were stably transduced with GFP-lentiviral vector enabling faster high throughput quantification of 3D microtissue growth assessment using an Operetta working Software and detection system ‘Harmony 3.0’ (PerkinElmer, USA). We studied the ability of RCTx to modify 3D-microtissue growth 3, 6 and 10 days after treatment. Results for five compounds (Actinomycin D, Staurosporine, Docetaxel, Doxorubicin and Vinblastine) showed that the IC50 values were improved by the addition of the single 2Gy radiation dose, indicating that they are capable of inducing a radiosensitation effect on radioresistant breast cancer cells. Panels of commercial secondary functional assays were adapted to the 3D-microtissue high throughput assay. Cellular viability and cytotoxicity were measured directly in microtissues using the CellTiter-Glo Reagent (Promega, USA). Apoptosis was measured using an ELISA based M30-Apoptosense assay (TECOmedical AG, Switzerland). These results all confirm that the assay operated with the 3D-microtissue model system is able to detect compounds that modulate tumors cell survival after irradiation.

Citation Format: Natasa Anastasov, Ines Höfig, Jan Lichtenberg, Simon Stroebel, Michael Salomon, Christian Thirion, Jens Kelm, Michael J. Atkinson. Identification of compounds modifying radiation-therapy using a 3D-microtissue technology. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5528. doi:10.1158/1538-7445.AM2013-5528
Abstract 5529: In vitro platforms representing tumor complexity for target validation and drug discovery.
Authors: Hickman John A.
Published: August 2014

PREDECT is one of the major Innovative Medicines Initiatives (IMI) of the European Union with 21 laboratories collaborating in a Public-Private partnership, aiming to create more appropriate in vitro platforms for target validation and drug discovery. Genomic and proteomic data on cancers from patient material provides hypotheses regarding potential novel drug targets. Laboratory platforms to validate whether target modulation would provide a clinical benefit are usually highly reductionist, often using long-established cell lines growing in 2 dimensions in vitro. These models do not reflect the complexity of a tumour in situ, where biochemical pathways are wired with connections to the complex tumour environment provided by stromal, immune and endothelial cells. PREDECT has the goal of comparing the pathological and molecular profiles of novel in vitro platforms with those of human tumours. In a stepwise manner, material from in situ tumours will be "deconstructed," for example through tissue slices, 3-D heterogenously complex models (with stroma etc), simple 3D models, complex 2D models to simple 2D models and the profiles of each platform compared with an in situ tumour, both in the resting state and when perturbed by drugs or RNA interference. "Reconstruction" of tumor complexity will also be attempted and profiled. Proteomics of models of breast, prostate and lung cancer will be compared in a Work Package based on a centralised TMA platform, immunohistochemistry and a Web-based microscopy. A systems approach will interrogate the fidelity of in vitro platforms to represent the complexity and heterogeneity of human tumors. Because clinical material presents both logistic and quality problems for ongoing and intense studies of target validation, PREDECT aims to use material from genetically engineered mouse models, and some "advanced" xenografts, whose pathology and molecular profiles closely match cohorts of human tumours. PREDECT hopes to provide more appropriate platforms both for target validation and subsequent preclinical studies which will replace a current cascade of tests which are poorly predictive of clinical activity.

Citation Format: John A. Hickman. In vitro platforms representing tumor complexity for target validation and drug discovery. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5529. doi:10.1158/1538-7445.AM2013-5529
Abstract 5590: Screening of pediatric and adult sarcoma cell lines reveals novel patterns of sensitivity toward approved oncology drugs and investigational agents.
Authors: August E. Michael , Delosh Rene , Laudeman Julie , Ogle Chad , Reinhart Russell , Selby Michael , Silvers Thomas , Morris Joel , Teicher Beverly A.
Published: August 2014

Sarcomas represent a heterogeneous group of neglected cancers with significant unmet medical need. To date, large-scale screening of compounds for activity against sarcomas has not been carried out in any systematic fashion to enable drug discovery. Our laboratory has compiled a panel of > 85 human pediatric and adult sarcoma cell lines, a tumor type not represented in the NCI60, and has carried out a high throughput screen of approximately 60 of these lines against the Approved Oncology Drugs (AOD) and Investigational Agents libraries, using inhibition of proliferation as an endpoint. The AOD library is comprised of 93 FDA-approved anticancer agents, and is available from The NCI/DTP Open Chemical Repository. The Investigational Agents library includes about 350 small molecules of interest. The compounds were screened for inhibition of cell proliferation in 9-point concentration response, starting at 10 uM and diluting by half-logs. The cells were cultured in 384-well plates for 24 hrs, at which time compounds were added with a final DMSO concentration of 0.25%. Viability was determined after 96 hrs of exposure using Alamar Blue. The % viability was determined relative to a vehicle-treated control, and this value was plotted relative to compound concentration to determine the EC50. The Ewing sarcoma lines (19) were unexpectedly more sensitive to pemetrexed, cabazitaxel, and floxuridine, and were, as expected, sensitive to topoisomerase 1 inhibitors relative to other sarcomas and to historical NCI60 data. Synovial sarcoma lines were sensitive to vinorelbine, vandetinib, and gemcitabine relative to other sarcomas. Further analyses and findings from the data will be presented. Funded by NCI Contract no. HHSN261200800001E.

Citation Format: E. Michael August, Rene Delosh, Julie Laudeman, Chad Ogle, Russell Reinhart, Michael Selby, Thomas Silvers, Joel Morris, Beverly A. Teicher. Screening of pediatric and adult sarcoma cell lines reveals novel patterns of sensitivity toward approved oncology drugs and investigational agents. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5590. doi:10.1158/1538-7445.AM2013-5590
Abstract 5599: A novel treatment strategy targeting shugoshin 1 in hematological malignancies.
Authors: Ikezoe Takayuki , Yang Jing , Nishioka Chie , Yokoyama Akihito
Published: August 2014

The shugoshin 1(SGOL1), a member of centromeric proteins, plays an important role in mitosis. The present study explored the levels of SGOL1 in hematological malignancies and found that SGOL1 was aberrantly expressed in various types of human leukemia cell lines (n=10, e.g., HL60, U937, MOLM-13, K562, EOL-1, etc.) and freshly isolated leukemia cells from individuals with acute myelogenous leukemia (AML, n=43, p<0.001) compared with bone marrow mononuclear cells isolated from healthy volunteers (n=9), as measured by real-time RT-PCR. Forced-expression of SGOL1 in hematopoietic stem/progenitor cells (HSPCs) significantly increased colony numbers for CFU-M and CFU-GM compared with control vector transduced infected HSPCs, suggesting that SGOL1 might act as an oncogene in hematopoietic cells. In addition, we found that repression of SGOL1 by a small interfering RNA (siRNA) slowed the proliferation of NB4, EOL-1 and U937 cells compared to the control siRNA transfected cells, in parallel with appearance of precocious dissociation of centromeric cohesion and separation of sister chromatids in these cells. Furthermore, we found that repression of SGOL1 by an siRNA accumulated EOL-1 and U937 cells in the G2/M phase of the cell cycle, in conjunction with up-regulation of spindle checkpoint protein BubR1, followed by apoptosis via caspase pathways. Taken together, SGOL1 may be a promising molecular target for individuals with AML.

Citation Format: Takayuki Ikezoe, Jing Yang, Chie Nishioka, Akihito Yokoyama. A novel treatment strategy targeting shugoshin 1 in hematological malignancies. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5599. doi:10.1158/1538-7445.AM2013-5599

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 5602: Cyclin D2- mediated cardiomyocyte cell cycle activity reverses doxorubicin-induced cardiotoxicity.
Authors: Zhu Wuqiang , Soonpaa Mark H. , Zhang Wenjun , Shou Weinian , Payne Mark , Field Loren J.
Published: August 2014

Introduction: Doxorubicin (DOX) is an effective anticancer agent which also induces acute and chronic cardiotoxicities. We have generated a juvenile mouse model to study DOX-induced cardiotoxicity in children. Cardiomyocyte apoptosis with an ensuing loss of myocardial function is observed in this model. Here, we hypothesize that activation of cardiomyocyte cell cycle protects against DOX-induced cardiotoxicity. Methods: Two week old MHC-cylin D2 (D2) mice and their non-transgenic (NON-TXG) littermates were given weekly DOX injections for 5 weeks (25 mg/kg total dose). Animals were studied 1 week (acute stage) or 13 weeks (late stage) after the last DOX injection. Cardiac function was measured with echocardiography and cardiomyocyte (CM) cell cycle was quantified using a tritiated thymidine incorporation assay. Results: Our results showed that CM cell cycle was increased in D2 mice as compared to NON-TXG mice (thymidine positive CM nuclei, 0.03±0.01% vs. 0.002±0.002 in acute stage, p<0.01; and 0.03±0.01% vs. 0.001±0.001 in late stage, p<0.01). In the absence of cell cycle activity, cardiac function remained depressed in late stage vs. acute stage (Fractional Shortening, FS, 41.4±3.1% vs. 44.0±3.2%, p>0.05), whereas in the presence of CM cell cycle activity, cardiac function improved in late stage vs. acute stage (FS = 60.1±1.3% vs. 47.1±3.3%, p<0.01). CM apoptosis (0.02±0.01/mm2 vs. 0.05±0.01/mm2, p<0.01) and fibrosis (6.2±0.9% vs. 12.0±1.7%, p<0.01) was reduced in D2 mice as compared to NON-TXG mice in late stage. Conclusion: For the first time, our data suggest activation of CM cell cycle in adult mice reverses DOX-induced cardiotoxicity.

Citation Format: Wuqiang Zhu, Mark H. Soonpaa, Wenjun Zhang, Weinian Shou, Mark Payne, Loren J. Field. Cyclin D2- mediated cardiomyocyte cell cycle activity reverses doxorubicin-induced cardiotoxicity. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5602. doi:10.1158/1538-7445.AM2013-5602
Abstract 5603: Minocycline abrogates IL-6 signaling and inhibits migration, invasion and adhesion capacity of ovarian cancer cells.
Authors: Ataie-Kachoie Parvin , Morris David L. , Pourgholami Mohammad H.
Published: August 2014

Aims: Minocycline (7-dimethylamino-6-desoxytertracycline) has been recognized as a drug with interesting pleiotropic properties including suppression of pro-inflammatory cytokine levels. The beneficial effects of minocycline arisen from suppression of proinflammatory cytokines and in particular IL-6 have been elucidated in central nervous system. However, this drug has never been examined in cancers with aberrant IL-6 expression. Considering the key role of IL-6 and its signaling pathways in the growth and progression of ovarian cancer, we evaluated whether minocycline modulates IL-6, its receptor system and signaling pathways in ovarian cancer.

Methods: Expression of IL-6 in control and minocycline-treated ovarian cancer cell lines (OVCAR-3, SKOV-3 and CAOV-3) was determined by immunocytochemistry. ELISA was used to quantify the effects of minocycline on both constitutive and IL-1β or 4-OH-estradiol stimulated IL-6 expression in these cell lines. The effects of minocycline on IL-6R expression was assessed by immunocytochemistry and western blot. Activation of STAT3 and ERK1/2, two major pathways down-stream of IL-6 were evaluated by western blotting. To further characterize the influence of minocycline against IL-6 bioactivity, the effect of the drug on the functional status of SKOV-3 cells was examined by adhesion assay and transwell migration and invasion assay. Finally, to determine the effect of minocycline on IL-6 expression in vivo, female nude mice bearing intraperitoneal OVCAR-3 tumors were treated with a single 30 mg/kg dose of minocycline administered intraperitoneally for 4 and 24 h. The IL-6 expression in plasma and tumor samples excised from these animals was evaluated by ELISA and western blot.

Results: We found that minocycline was potent in suppressing both constitutive and stimulated IL-6 expression in vitro. Analysis of plasma and tumors also revealed substantial down-regulation of IL-6 in minocycline-treated mice in vivo. Moreover, minocycline blocked the expression of IL-6R and phosphorylation of STAT3 and ERK1/2 time-dependently. It was also found that minocycline-treated cells exhibit dose-dependent attenuation of cell adhesion, migration and invasion capacity.

Conclusion: These results demonstrated that minocycline effectively blocks IL-6 signaling in ovarian cancer. Blockage of IL-6 signaling may provide benefits for the treatment of this deadly cancer.

Citation Format: Parvin Ataie-Kachoie, David L. Morris, Mohammad H. Pourgholami. Minocycline abrogates IL-6 signaling and inhibits migration, invasion and adhesion capacity of ovarian cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5603. doi:10.1158/1538-7445.AM2013-5603
Abstract 5610: Electric-field assisted delivery of gemcitabine for the treatment of pancreatic cancer.
Authors: Byrne James , O'Neill Adrian , Jajja Raheel , Napier Mary , Luft James C. , Zamboni William , Yeh Jen Jen , DeSimone Joseph
Published: August 2014

Pancreatic cancer is the fourth leading cause of cancer death in the world, with an incidence rate approximately equivalent to the death rate. The very poor prognosis for pancreatic cancer can be attributed to late diagnosis and the ineffectiveness of drug delivery to the primary tumor. Poor tissue perfusion plays a substantial role in preventing adequate drug exposure to primary pancreatic tumors. The systemic administration of gemcitabine, the current standard-of-care chemotherapy for pancreatic cancer, has shown limited efficacy for the treatment of pancreatic cancer, with only 5 to 10% of patients demonstrating an objective radiographic response at the primary tumor site. Furthermore, the toxicity associated with systemic chemotherapy has been found to reduce the quality of life of the patient and in extreme cases can be fatal. In an attempt to address the lack of effectiveness of systemically administered chemotherapy and associated toxicity, we have developed modalities for the localized delivery of chemotherapies to pancreatic tumors. These devices rely upon an applied electric potential difference between electrodes to drive chemotherapy into the tumor. This use of an electric potential for local delivery of chemotherapies offers the capability of overcoming considerable flow and pressure gradients. Significant work using an applied electric potential difference for drug transport has been performed in transdermal drug delivery. In the area of oncology, electric field-assisted delivery techniques have been proposed for skin and ocular cancers and have been clinically translated to the treatment of bladder cancers in the electromotive delivery of mitomycin C. This technique, combined with novel device approaches, is particularly well suited for the local treatment of the primary pancreatic tumor. Herein, we report the development of electric field-assisted delivery devices and the testing of these devices in a patient-derived orthotopic xenograft mouse model of pancreatic adenocarcinoma and a healthy canine animal model. The results show that our devices provide a significant increase in the local delivery of gemcitabine while limiting systemic toxicity.

Citation Format: James Byrne, Adrian O'Neill, Raheel Jajja, Mary Napier, James C. Luft, William Zamboni, Jen Jen Yeh, Joseph DeSimone. Electric-field assisted delivery of gemcitabine for the treatment of pancreatic cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5610. doi:10.1158/1538-7445.AM2013-5610
Abstract 5613: Enhanced treatment of lung metastasis of triple negative breast cancer by doxorubicin-mitomycin C co-loaded polymer lipid nanoparticles.
Authors: Prasad Preethy , Cai Ping , Rauth Andrew , Wu Xiao
Published: August 2014

Background: Treatment of triple negative breast cancer (TNBC) is a big challenge due to its aggressiveness, metastases, lack of therapeutic target and receptor proteins. Our laboratory has developed doxorubicin (Dox) -mitomycin C (MMC) co-loaded stealth polymer lipid hybrid nanoparticles (DMsPLN) and demonstrated their anti-cancer synergy in vitro and high efficacy in vivo. The purpose of this study is to evaluate the biodistribution, in vivo efficacy and safety of DMsPLN in a lung metastatic human TNBC model.

Method: Lung metastasis of breast tumor was established using MDA-MB 231-luc-D3H2LN, a luciferase expressing cell line that was derived from spontaneous lymph node metastasis. The cells were injected via tail vein to develop lung metastasis in SCID mice. The biodistribution and tumor accumulation of the nanoparticles were examined by whole animal optical imaging using near infrared fluorescence labeled nanoparticles. The efficacy and systemic toxicity of DMsPLN were evaluated against clinically used doxorubicin (Dox). DMsPLN and free Dox solution were administered intravenously at various equivalent Dox doses. The tumor burden was monitored weekly by bioluminescence imaging. The size and number of tumor nodules in the lungs were examined at different time points and the systemic toxicity was monitored by repeated measurement of body weight.

Results: A lung metastatic breast tumor model was successfully developed using the MDA-MB 231-luc-D3H2LN which allowed for non-destructive monitoring of tumor growth and metastases using bioluminescence imaging. Whole animal imaging demonstrated the accumulation of the fluorescent nanoparticles in the lung metastatic site. Treatment with free Dox (10 mg/kg) resulted in severe total body weight loss over 20% which is ruled as a toxic endpoint. However, the DMsPLN (10 mg/kg equivalent Dox dose) group did not show any systemic toxicity. Treatment with DMsPLN at various Dox doses resulted in a reduction in tumor burden compared to the saline and Dox-treated groups.

Citation Format: Preethy Prasad, Ping Cai, Andrew Rauth, Xiao Wu. Enhanced treatment of lung metastasis of triple negative breast cancer by doxorubicin-mitomycin C co-loaded polymer lipid nanoparticles. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5613. doi:10.1158/1538-7445.AM2013-5613

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 5614: Legumain protease substrate modified TAT-liposome cargo: an efficient tool for targeting malignant diseases.
Authors: Liu Ze , Xiong Min , Liao Debbie , Lv Dan , Reisfeld Ralph A. , Wrasidlo Wolfgang , Chen Si , Stupack Dwayne G. , Sun Peiqing , Tan Xiaoyue , Xiang Rong
Published: August 2014

Tumor targeting specificity and efficacy of the internalization of macro-biomolecular drugs have emerged as two major obstacles hampering the application of liposome-based delivery system against tumor. Trans-activating transcriptional activator (TAT) is a member of the cell-penetrating peptides which enhance the penetration of various macro-biomolecular cargos. In this study, we developed a modified TAT-Liposome cargo in which the activation of TAT peptide was blocked by Legumain protease substrate Ala-Ala-Asn-(AAN). Firstly, we confirmed the extremely high level of Legumain expression in a variety of solid tumors, as well as its specific effect on recognition and digestion of AAN. Then we conjugated AAN with the TAT peptide and constructed the modified liposome (AAN-TAT-Lipo) cargo. Flow-cytometry assay showed the internalization of AAN-TAT-Lipo was significantly higher than that of AAN-Lipo in 4T1 cells which expressed active Legumain. In vivo, the concentration of Dox encapsulated in AAN-TAT-Lipo was significantly higher in the tumor sites and lower in normal tissues compared with Free-Dox, AAN-Lipo-Dox and TAT-Lipo-Dox groups after administration in mouse model of orthotopic breast cancer. Luciferase assay showed that the AAN-TAT-Lipo-Dox dramatically inhibited tumor growth compared with other groups. TUNEL staining demonstrated that the apoptotic cells in tumor sites were significantly increased in the group of AAN-TAT-Lipo-Dox, compared with other groups. Whereas, the apoptotic cells in the heart, liver, spleen and kidney were decreased in the group of AAN-TAT-Lipo-Dox. Taken together, the modified TAT-lipo cargo via blocking TAT peptide activation with a Legumain specific substrate, AAN, is proven effective for improving the efficacy of internalization, and reducing the systemic toxicity of the tumor targeting delivery system as well.

Citation Format: Ze Liu, Min Xiong, Debbie Liao, Dan Lv, Ralph A. Reisfeld, Wolfgang Wrasidlo, Si Chen, Dwayne G. Stupack, Peiqing Sun, Xiaoyue Tan, Rong Xiang. Legumain protease substrate modified TAT-liposome cargo: an efficient tool for targeting malignant diseases. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5614. doi:10.1158/1538-7445.AM2013-5614
Abstract 5634: Increased co-amplification of HER2 and STARD3 in a cell line model of acquired lapatinib resistance.
Authors: McDermott Martina , Anderson Lee , Shields Liam , O'Brien Neil , Prendergast Allison , Kennedy Susan , Gallagher William , Zagozdzon Radoslaw , Byrne Annette , Crown John , Slamon Dennis , O'Donovan Norma
Published: August 2014

Resistance to lapatinib, a tyrosine kinase inhibitor of HER2, is a significant clinical problem and to date the mechanisms of acquired lapatinib resistance remain largely unknown. In order to study acquired lapatinib resistance we established an in vitro cell line model through continuous culture of HCC1954 cells with 1 μM lapatinib. After 6 months the resulting HCC1954-L cells where significantly less sensitive to lapatinib with an IC50 of 2.6 ± 0.3 μM compared to aged-matched parental cells (IC50 0.42 ± 0.02 M; p = 0.004). In addition, lapatinib treatment had no significant effect on the growth of HCC1954-L xenograft tumors in mice, indicating the utility of HCC1954-L cells as an in vivo model of acquired lapatinib resistance.

Array CGH analysis was performed on the sensitive and resistant cell lines. Chromosomal aberrations were detected in 15 regions in the HCC1954-L cells, including increased amplification of the region of chr17q12 known to contain HER2 and STARD3. Immunoblotting was performed to assess whether the increased amplification corresponded with increased expression of the proteins. HCC1954-L cells exhibited a small but significant increase (1.2 fold) in HER2 protein expression (p=0.02) compared to parental cells. Increased gene amplification of HER2 was also confirmed by FISH analysis. While STARD3 protein expression was undetectable in in parental HCC1954 cells, HCC1954-L cells expressed high levels of STARD3 protein. Knockdown of STARD3 using siRNA resulted in a 20% decrease in cell growth compared to scrambled control treated cells, suggesting that STARD3 amplification and expression plays a role in the proliferation of HCC1954-L cells.

In summary, we have developed a robust model of acquired lapatinib resistance which exhibits increased amplification and expression of HER2 and STARD3.

Citation Format: Martina McDermott, Lee Anderson, Liam Shields, Neil O'Brien, Allison Prendergast, Susan Kennedy, William Gallagher, Radoslaw Zagozdzon, Annette Byrne, John Crown, Dennis Slamon, Norma O'Donovan. Increased co-amplification of HER2 and STARD3 in a cell line model of acquired lapatinib resistance. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5634. doi:10.1158/1538-7445.AM2013-5634
Abstract 5645: Angiogenic activity leads failure of antitumor activity of cixutumumab, an anti-Insulin like growth factor receptor monoclonal antibody.
Authors: Lee Ji-Sun , Kang Ju-Hee , Boo Hye-Jin , Myers Jeffrey N. , Johnson Faye M. , Glisson Bonnie S. , Oh Seoung-Hyun , Lee Ho-Young
Published: August 2014

The type 1 insulin-like growth factor receptor (IGF-1R) signaling is dysregulated in many cancers, thus being a target of cancer therapy. Several preclinical trials have reported the efficacy of IGF-1R targeted therapies, mostly using tyrosine kinase inhibitors or monoclonal antibodies (mAbs); however many of clinical trials have shown limited therapeutic efficacy. In this study, we investigated the resistance mechanism to IGF-1R mAb-based therapies using cixutumumab, a fully humanized mAb against IGF-1R. We tested the efficacy of cixutumumab using OSC19 and 686LN head and neck squamous cell carcinoma (HNSCC) and MDA-MB-231 breast cancer orthotopic and H1299 non-small cell lung cancer (NSCLC) xenograft tumor models. We observed that cixutumumab treatment induced either no change or increased tumor growth and decreased the survival duration of the animals bearing those tumors. Treatment with cixutumumab upregulated migration and invasion of HNSCC and NSCLC cells without any effects on cell proliferation. We further examined the effects of cixutumumab on tumor angiogenesis and found that conditioned media from cancer cells treated with cixutumumab increased tube formation and migration of human umbilical endothelial cells (HUVECs). The angiogenic activity of cixutumumab was also confirmed by matrigel plug assay with HNSCC cell lines. Collectively, these data suggest that cixutumumab-induced angiogenesis-stimulating activities and migration and invasion potential of cancer cells are major factors of resistance to IGF-1R mAbs. These data further suggest that inhibitors of angiogenesis are useful strategy to overcome the resistance against the IGF-1R mAb-based therapy.

Citation Format: Ji-Sun Lee, Ju-Hee Kang, Hye-Jin Boo, Jeffrey N. Myers, Faye M. Johnson, Bonnie S. Glisson, Seoung-Hyun Oh, Ho-Young Lee. Angiogenic activity leads failure of antitumor activity of cixutumumab, an anti-Insulin like growth factor receptor monoclonal antibody. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5645. doi:10.1158/1538-7445.AM2013-5645
Abstract 5661: Nanoparticle based combinatorial siRNA therapy against human hepatocellular carcinoma (HCC) .
Authors: Decaens Thomas , Factor Valentina M. , Kulic Iva , Andersen Jesper B. , Seo Daekwan , Lee Yun-Han , Judge Adam D. , Conner Elizabeth A. , MacLachlan Ian , Thorgeirsson Snorri S.
Published: August 2014

Background: We have previously demonstrated the therapeutic effect of lipid nanoparticles (LNP) loaded with single siRNA targeting CSN5 and WEE1 against human HCC in mouse models.

Aim: To test the benefit of a combinatorial versus single siRNA therapy in mouse models of human HCC and to identify molecular mechanism(s) involved in therapeutic response by extensive microarray analyses.

Materials and Methods: LNP formulations of chemically modified siRNAs targeting CSN5 and WEE1 were produced by Tekmira® Pharmaceuticals. SCID-beige mice were used for subcutaneous (Hep3B) and intra-hepatic (Huh7-luciferase) tumor transplantation. Mice with established tumors were treated intravenously with 4 mg/kg single agent siRNA or 2 mg/kg + 2 mg/kg siCSN5:siWEE1 siRNA co-encapsulated in the same LNP. Tumors were assayed following 1 to 9 injected doses. Tumor progression in the Huh7 orthotopic model was monitored by bioluminescence imaging and metastases were evaluated at endpoint.

Results: Significant target silencing was observed in tumors after single or repeat administration with no antagonism between siRNAs occurring in the CSN5:WEE1 combination. We observed significant inhibition of tumor growth and metastases in mice treated with active siRNAs compared to LNP containing a non-targeting control sequence. Potency was not lost with siCSN5:siWEE1 LNP, where the concentration of each siRNA is halved in combination, relative to the most efficacious single agent. Data from preliminary microarray analyses demonstrate a strong transcriptome difference between each treatment group.

Conclusion: We demonstrate a clear efficacy of a LNP based combinatorial siRNA therapy in human mouse models of HCC. Overall this therapy led to a significant decrease of tumor size induced by mRNA inhibition

Citation Format: Thomas Decaens, Valentina M. Factor, Iva Kulic, Jesper B. Andersen, Daekwan Seo, Yun-Han Lee, Adam D. Judge, Elizabeth A. Conner, Ian MacLachlan, Snorri S. Thorgeirsson. Nanoparticle based combinatorial siRNA therapy against human hepatocellular carcinoma (HCC) . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5661. doi:10.1158/1538-7445.AM2013-5661
Abstract 5663: In vitro radiosensitization of human cervical cancer HeLa cells with iron oxide- titanium dioxide core-shell nanoparticles conjugated with 3,4-Dihydroxyphenylacetic acid (DOPAC).
Authors: Mirzoeva Salida , Wanzer Beau M. , Paunesku Tatjana , Woloschak Gayle E.
Published: August 2014

Cervical cancer is resistant to current surgical, chemo- and radiotherapy interventions and represents a virtually incurable tumor. In this study, we explored titanium dioxide core-shell nanocomposites conjugated with DOPAC (DOPAC-NCs) as radiosensitizers. Radioresistant human carcinoma cervical HeLa cell cultures were treated with or without of increasing concentrations of DOPAC-NCs (5-20 nM) prior to irradiation. One hour later, cells were irradiated with a single dose of gamma radiation (5 Gy, 10 Gy, or 15 Gy). Seventy-two hours following radiation treatment, cell proliferation/viability was assessed by MTS assay. Sham-irradiated cells were served as a control. We have found that treatment of HeLa cells with DOPAC-NCs affected the survival of irradiated cells in a dose-dependent manner. Our data demonstrate that cell survival of cells irradiated with 15 Gy and pre-treated with the highest concentration of DOPAC-NCs was as low as 23.7% compared to survival of cells that were not treated with DOPAC-NCs prior to irradiation. In opposite, without radiation, treatment with DOPAC-NCs had no effect on cell viability/proliferation.

These findings provide in vitro evidence that DOPAC-NCs may be developed and employed as radiosensitizers in cervical cancer radiation therapy.

Citation Format: Salida Mirzoeva, Beau M. Wanzer, Tatjana Paunesku, Gayle E. Woloschak. In vitro radiosensitization of human cervical cancer HeLa cells with iron oxide- titanium dioxide core-shell nanoparticles conjugated with 3,4-Dihydroxyphenylacetic acid (DOPAC). [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5663. doi:10.1158/1538-7445.AM2013-5663
Abstract 5668: An experimental study of folic acid coupled nano-paclitaxel for targeted treatment of ovarian cancer.
Authors: Li Hongxia
Published: August 2014

To study the effect of folic acid coupled nano-paclitaxel in vivo and in vitro targeted treatment of ovarian cancer and preliminarily elucidate its therapeutical mechanism. Methods: The folic acid coupled nano-paclitaxel drug were prepared by the co-incubation method; The folic acid receptor-positive ovarian cancer cell lines were screened by immunohistochemical staining method; the targeting effect of the drug on SKOV3 cells were assessed qualitatively by fluorescence microscopy and quantitatively by flow cytometry (FCM). The killing effect of the drug on SKOV3 cells and SKOV3/TAX cells were monitored in vitro by plotting cell growth curve, the MTT method, FCM measurement, and electron microscopy; Tumor-burdened nude mice were administered with the drug by intraperitoneal and iv injection, the amounts of tumors were compared. The tumor killing effect of the drug in treating celiac metastases were examined qualitatively by electron microscopy and quantitatively by FCM. Results: (1) The particle size and encapsulation efficiency of folic acid coupled nano-paclitaxel drugs were (140.5±10.3)nm and 97% respectively; (2) Both SKOV3 cells and SKOV3/TAX cells were folic acid receptor positive; (3) Folic acid coupled nano-paclitaxel drugs demonstrated targeted killing effect on SKOV3 cells by the mediation of the folic acid receptor. The intracellular drug concentration gradually increases along with the incubation time. (4)The effects of folic acid coupled nano-paclitaxel drugs on SKOV3 cells and SKOV3/TAX cells were significantly better than the nano-paclitaxel, but the effects was similar when the concentrations were 0.2 and 2 μg/ml. The resistance index of SKOV3 / TAX cells was decreased in the course of treatment; (5) The cell apoptosis rates of the three groups after treatment were 84.63% in abdominal group,35.23% in veinal group,and 10.12% in control group respectively (p < 0.01). The apoptotic necrosis in tumor issues of abdominal group was quite obviously. But the tumor-burdened quantities in three groups were 3.38±1.77, 4.50±2.20, and 3.86±1.95 respectively. there were no statistical significance (P > 0.05). Conclusions: Folic acid coupled nano-paclitaxel drugs have better therapeutical effects in treating both drug-sensitive and drug-resistant ovarian cancer lines. The drugs administered through intraperitoneal injection showed better results than via iv injection.

Citation Format: Hongxia Li. An experimental study of folic acid coupled nano-paclitaxel for targeted treatment of ovarian cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5668. doi:10.1158/1538-7445.AM2013-5668

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 5678: Design of GRP78 inhibitors as novel therapeutics for breast cancer.
Authors: Ramkumar Kavya , Debnath Bikash , Lee Amy S. , Neamati Nouri
Published: August 2014

Cancer cells exploit cellular stress response mechanisms, such as the unfolded protein response, to survive through cytotoxic treatments. A key mediator of the unfolded protein response, the 78kDa glucose regulated protein (GRP78) chaperone, has frequently been found to be overexpressed in breast cancer. In addition to providing a growth advantage to cancer cells, GRP78 induction leads to drug resistance. Suppressing GRP78 levels using siRNA has been shown to slow cancer cell progression and overcome drug resistance. Another approach to inactivating GRP78 is by inhibiting its ATPase activity. Currently, there are only few selective GRP78 inhibitors in development despite ample evidence of its critical role in cancer cell survival and drug resistance. Here, we report a novel class of small-molecule inhibitors of GRP78 ATPase activity. We designed a focused library of compounds based on a structure-based docking approach and screened for inhibition of GRP78 ATPase activity. Based of the initial hit, we further optimized several analogues with more potent activity and selectivity for GRP78 inhibition (IC50 <5μM). The lead compound, KR-240, induced CHOP significantly under conditions of ER stress and activated the pro-apoptotic arm of the unfolded protein response. KR-240 showed moderate cytotoxicity against a panel of breast cancer cell lines and increased sensitivity to chemotherapeutic agents, further supporting the role for GRP78 in cancer survival and drug resistance. Overall, our results indicate that our novel GRP78 inhibitor, KR-240 has the potential for further development as an anticancer agent.

Citation Format: Kavya Ramkumar, Bikash Debnath, Amy S. Lee, Nouri Neamati. Design of GRP78 inhibitors as novel therapeutics for breast cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5678. doi:10.1158/1538-7445.AM2013-5678
Abstract 572: Proline-rich acidic protein 1 (PRAP1), a novel mitotic arrest deficient 1 (MAD1) interacting partner, suppresses mitotic checkpoint signaling in hepatocellular carcinoma.
Authors: Sze Karen MF , Chu Glanice K. , Mak Queenie HY , Ng Irene OL
Published: August 2014

DNA aneuploidy or chromosomal instability is commonly observed in major types of human cancers, and one of the causes for chromosomal instability is defective mitotic checkpoint signaling in cells. Loss of mitotic checkpoint contributes to chromosomal instability, leading to carcinogenesis. The mitotic checkpoint monitors the status of kinetochore-microtubule attachment and delays the onset of anaphase until all kinetochores have been attached with microtubules. Mitotic arrest deficient 1 (MAD1) is key component in mitotic checkpoint signaling. We previously identified a novel isoform of MAD1, which we named MAD1β (Sze et al. Cancer Res 2008;68:9194-9201). In this study, using yeast-two hybrid screening, we have identified a novel MAD1 interacting partner, proline-rich acidic protein 1 (PRAP1). The function of PRAP1 is largely unknown. Hence, we investigated its role in mitotic checkpoint signaling in hepatocellular carcinoma (HCC). We demonstrated the physical interaction between PRAP1, MAD1 and MAD1β in yeast-two hybrid screening and with co-immunoprecipitation assay. Both MAD1 and MAD1β proteins were co-immunoprecipitated with PRAP1. Moreover, we stably expressed PRAP1 in two HCC cell lines, BEL-7402 and SMMC-7721, which have competent mitotic checkpoints. We observed that stable expression of PRAP1 induced impairment in mitotic checkpoint (P < 0.01). After treatment with either colcemid or nocodazole, the PRAP1-stably expressing BEL-7402 and SMMC-7721 cells showed significantly lower mitotic indices as compared with the corresponding vector control cells. Ectopic expression of PRAP1 also resulted in chromosome bridge formation (P< 0.01) and aberrant chromosome numbers (P < 0.001) in these cell lines. All these cellular changes contributed to chromosomal instability. Interestingly, ectopic expression PRAP1 led to significant underexpression of MAD1 but not MAD1β in HCC cells. Taken altogether, our data indicate that PRAP1 is an interacting partner of MAD1 and may have functions of degrading MAD1 and in turn suppressing mitotic checkpoint signaling in HCC.

(This study was partly funded by a Hong Kong Research Grants Council GRF grant [HKU 772608])

Citation Format: Karen MF Sze, Glanice K. Chu, Queenie HY Mak, Irene OL Ng. Proline-rich acidic protein 1 (PRAP1), a novel mitotic arrest deficient 1 (MAD1) interacting partner, suppresses mitotic checkpoint signaling in hepatocellular carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 572. doi:10.1158/1538-7445.AM2013-572