bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5214: miR-424(322)/503 cluster orchestrates remodeling of the epithelium in the involuting mammary gland and plays a role in breast cancer development
Authors: Navas David Llobet , Barrueco Ruth Rodriguez , Silva Jose
Published: May 2015

The mammary gland is a very dynamic organ that undergoes continuous remodeling. The critical regulators of this process are not fully understood. Here we identify the microRNA cluster miR-424(322)/503 as an important regulator of epithelial involution after pregnancy and tumorigenesis. Importantly, miR-424(322)/503 loci is deleted in 20% of basal and luminal B tumors. Through the generation of a knock-out mouse model, we found that regression of the secretory acini of the mammary gland was compromised in the absence of miR-424(322)/503. Mechanistically, we show that miR-424(322)/503 orchestrates cell life and death decisions by targeting CDC25A, BCL-2 and IGF1R. Furthermore, we demonstrate that the expression of this miR-cluster is regulated by TGF-β, a well characterized regulator of mammary involution. Overall, our data suggest a model where activation of the TGF-β pathway after weaning induces the transcription of the miR-424(322)/503 which in turn downregulates the expression of key genes. Here, we unveil a previously unknown, multilayered-regulation of epithelial tissue remodeling coordinated by the microRNA cluster miR-424(322)/503.

Note: This abstract was not presented at the meeting.

Citation Format: David Llobet Navas, Ruth Rodriguez Barrueco, Jose Silva. miR-424(322)/503 cluster orchestrates remodeling of the epithelium in the involuting mammary gland and plays a role in breast cancer development. [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 5214. doi:10.1158/1538-7445.AM2014-5214
Abstract 522: Revealing the complexity of cancer associated small non-coding RNAs by next generation sequencing (NGS) and low-density array
Authors: Montani Francesca , Marzi Matteo , Carletti Rose Mary , Fiore Pier Paolo Di , Nicassio Francesco
Published: May 2015

Background. The recent development of high-throughput sequencing technologies (Next Generation Sequencing - NGS) provided instruments to reveal the complexity of nucleic acids, highlighting the existence of numerous species of non-coding RNAs (ncRNAs) that play a significant regulatory role in complex organisms, impacting on both physiology and disease. In particular, microRNAs (miRNAs) have been proposed as “next-generation” markers for their ability to unambiguously distinguish cellular states, including SCs, progenitors and differentiated cells, as well as tumour types, even among closely related cancers or in different biological fluids. However, a comprehensive approach for the identification and characterization of small non-coding RNAs from pathological samples (blood, FFPE, primary cultures) has not been established nor compared with current available methodologies (i.e. QPCR).

Aim: Our aim is to develop a simultaneous and comparative protocol for the analysis of small non-coding RNAs from pathological samples, which encompasses both high-throughput QPCR and sequencing analysis in order to reveal the complexity of cancer associated small ncRNAs.

Results: An optimized protocol to analyse small ncRNAs from different pathological samples has been developed and compared with high-throughput QPCR. Both platforms resulted as highly efficient and quantitative, although NGS manages to score many molecules and RNA species that could not be analysed by current QPCR platforms, thus revealing a major complexity of cancer associated non-coding RNAs.

Conclusion: We propose that the combined use of NGS and QPCR platforms would allow a wider and more detailed analysis of ncRNAs, expanding our ability to fish out robust and efficient molecular markers for diagnostic (early detection), prognostic or therapeutic use.

Note: This abstract was not presented at the meeting.

Citation Format: Francesca Montani, Matteo Marzi, Rose Mary Carletti, Pier Paolo Di Fiore, Francesco Nicassio. Revealing the complexity of cancer associated small non-coding RNAs by next generation sequencing (NGS) and low-density array. [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 522. doi:10.1158/1538-7445.AM2014-522
Abstract 5229: Profiling of circulating miRNAs is a critical prognostic biomarker tool for high-risk neuroblastoma
Authors: Ramraj Satish Kumar , Pandian Vijayabaskar , Khan Faizan H. , Aravindan Sheeja , Herman Terence S. , Natarajan Mohan , Aravindan Natarajan
Published: May 2015

Circulating miRNAs has momentous clinical relevance and has been correlated with disease progression in many solid tumors. In this study, we investigated the circulating miRNAs' variations exploiting unique mouse models of non-metastatic favorable and high-risk, aggressive human neuroblastoma. Conditioned serum samples from animals bearing favorable or high-risk disease were examined for alterations in circulating miRNAs using whole genome human miRNA profiling approach. Serum from sham mice were used as mouse-background false-positive controls. Data after chip-to-chip normalization and mouse background subtraction were compared by adopting favorable-to-high-risk animal(s) crisscross analysis. Compared to favorable neuroblastoma, miRNA profiling revealed a significant (>2 fold) upregulation of 33 miRNAs and complete suppression of 42 miRNAs in animals with high-risk NB. Preferential exclusion of mouse miRNA with sequence homology blast revealed 11 (off 33 upregulated) unique human miRNA including miR-1261, miR-1268, miR-1280, miR-1304, miR-1308, miR-1908, miR-198, miR-513a-5p, miR-513b, miR-548h, miR-580. Consistently, homology blasting identified 13 (off 42 downregulated) miRNAs including, miR-1206, miR-548a-5p, miR-548f, miR-576-5p, miR-600, miR-639, miR-640, miR-641, miR-647, miR-662, miR-886-3p, miR-887, miR-888. Expression pattern of select miRNAs were validated using QPCR. Target analysis for these miRNAs revealed a signature pattern, mostly pertaining to stemness and tumor progression flow-through signaling viz., NKX3.2, MEGF10, GRB10, GPX8, ZEB1, SIX-1, ZIC2, MMP-2, SMAD4, KRAS, NFkB1, CD24, CD44. Together, these data identifies unique circulating miRNA signature of high-risk neuroblastoma and further implies that these miRNAs may play functional role in the progression of the disease.

Citation Format: Satish Kumar Ramraj, Vijayabaskar Pandian, Faizan H. Khan, Sheeja Aravindan, Terence S. Herman, Mohan Natarajan, Natarajan Aravindan. Profiling of circulating miRNAs is a critical prognostic biomarker tool for high-risk neuroblastoma. [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 5229. doi:10.1158/1538-7445.AM2014-5229
Abstract 5358: IL-4/IL-13 induce Duox2/DuoxA2 expression and reactive oxygen production in human pancreatic and colon cancer cells
Authors: Wu Yongzhong , Doroshow James H.
Published: May 2015

NADPH oxidase (NOX)-derived reactive oxygen species (ROS) contribute significantly to inflammation-associated carcinogenesis. Expression of dual oxidase 2 (Duox2), one of seven members of the NOX gene family, is up-regulated in inflammatory bowel disease, chronic pancreatitis, and in many human malignancies including carcinomas of the prostate, lung, and breast. Previously, we demonstrated that Stat1 and/or NF-κB play a critical role in modulating the enhanced expression of Duox2, and its cognate maturation factor DuoxA2, by IFN-γ and lipopolysaccharide in human pancreatic cancer cells. This cytokine-mediated increase in Duox2 expression is responsible for a consequent significant increase in tumor cell H2O2 production and DNA damage. Using Caco2 and T84 human colon cancer lines, and BxPC-3 pancreatic cancer cells, we now report that two other pro-inflammatory cytokines, IL-4 and IL-13, also strongly induce Duox2/DuoxA2 RNA and protein expression (>5-10-fold after 24h treatment with 50 ng/ml cytokine). In the BxPC-3 line, IL-4 exposure activates the Stat6 signaling pathway, enhances expression of Duox2/DuoxA2 in a time- and concentration-dependent manner, and leads to a significant increase in the generation of extracellular H2O2. The transcription and translation inhibitors actinomycin D and cyclohexamide both individually suppress IL-4-induced Duox2 expression, suggesting that IL-4-mediated Duox2 up-regulation is transcriptionally regulated, but that new protein synthesis is also necessary. We found that Stat6 is essential for IL-4-induced Duox2 expression because silencing Stat6 with Stat6-specific siRNA significantly attenuates IL-4-induced Duox2 RNA and protein levels in BxPC-3 cells. To evaluate the potential role of Duox2 in inflammation-associated human cancers further, immunohistochemical studies were performed to semi-quantitatively examine Duox expression using human pancreatic and colon cancer tissue arrays. We found that a majority of samples of pancreatic intra-epithelial neoplasia and frank pancreatic cancer demonstrated high-level expression of Duox , often in association with inflammatory cell infiltrates; high-level Duox expression was also found in most colorectal cancer samples. These studies, in concert with our previous experiments, strongly suggest that ROS, produced as a consequence of cytokine-induced Duox up-regulation, may be associated with the development of inflammation-related gastrointestinal malignancies.

Note: This abstract was not presented at the meeting.

Citation Format: Yongzhong Wu, James H. Doroshow. IL-4/IL-13 induce Duox2/DuoxA2 expression and reactive oxygen production in human pancreatic and colon cancer 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 5358. doi:10.1158/1538-7445.AM2014-5358
Abstract 5387: Dual kinase/bromodomain inhibitors for rationally designed polypharmacology
Authors: Quinn Elizabeth R. , Ciceri Pietro , Müller-Knapp Susanne , O'Mahony Alison , Fedorov Oleg , Filippakopoulos Panagis , Hunt Jeremy P. , Lasater Elisabeth A. , Pallares Gabriel , Picaud Sarah , Wells Christopher , Wodicka Lisa M. , Shah Neil P. , Knapp Stefan , Treiber Daniel K.
Published: May 2015

Concomitant inhibition of multiple cancer-driving kinases is an established strategy to improve the durability of clinical responses to targeted therapies. The difficulty of discovering kinase inhibitors with an appropriate multi-target profile has, however, necessitated the application of combination therapies, which can pose significant clinical development challenges. Epigenetic reader domains of the bromodomain family have recently emerged as novel targets for cancer therapy. Here we have used BROMOscan™ bromodomain ligand binding assays to identify several clinical kinase inhibitors that also inhibit bromodomains with therapeutically relevant potencies and are best classified as dual kinase/bromodomain inhibitors. Nanomolar activity on BRD4 by clinical PLK1 and JAK2/FLT3 kinase inhibitors is particularly noteworthy as these combinations of activities on independent oncogenic pathways exemplify a novel strategy for rational single agent polypharmacological targeting. Importantly, cell-based data show that these dual inhibitors suppress c-Myc expression (a hallmark of BRD4 inhibition) and induce complex polypharmacological phenotypes reflecting dual kinase/bromodomain inhibition across a panel of primary human cell assay systems that model complex tissue and disease state environments (BioMap™). Furthermore, rich structure-activity relationships for related inhibitors and co-crystal structures identify design features that enable a general platform for the rational design of dual kinase/bromodomain inhibitors.

Citation Format: Elizabeth R. Quinn, Pietro Ciceri, Susanne Müller-Knapp, Alison O'Mahony, Oleg Fedorov, Panagis Filippakopoulos, Jeremy P. Hunt, Elisabeth A. Lasater, Gabriel Pallares, Sarah Picaud, Christopher Wells, Lisa M. Wodicka, Neil P. Shah, Stefan Knapp, Daniel K. Treiber. Dual kinase/bromodomain inhibitors for rationally designed polypharmacology. [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 5387. doi:10.1158/1538-7445.AM2014-5387
Abstract 5390: Development of a high-throughput screening assay to identify UHRF1 inhibitors via time-resolved fluorescence resonance energy transfer (TR-FRET)
Authors: Walker David A. , Wyhs Nicolas , Giovinazzo Hugh , Yegnasubramanian Srinivasan , Nelson William G.
Published: May 2015

Aberrant DNA methylation of gene promoters can be involved in silencing tumor suppressor genes in cancer. The protein ubiquitin-like containing PHD and ring finger domains 1 (UHRF1) is an essential component in the cellular machinery for DNA methylation maintenance and can couple DNA methylation and other epigenetic histone modifications. UHRF1 is thought to bind newly synthesized hemimethylated DNA and recruit DNA methyltransferases to copy the methylation pattern to the daughter DNA strand. Interfering with the ability of UHRF1 to bind methylated DNA can lead to the re-expression of epigenetically silenced genes including tumor suppressors, which positions UHRF1 as a target for anticancer drug development. We have created a high-throughput, time-resolved fluorescence resonance energy transfer (TR-FRET)-based assay to screen for inhibitors capable of disrupting the interaction between the UHRF1 SRA domain and hemi-methylated DNA. The assay measures the degree of TR-FRET between a 5′-fluorescein (FAM) labeled hairpin oligonucleotide and a terbium-conjugated antibody bound to the 6xHis-tag of the UHRF1_SRA domain polypeptide. In 384-well plate format, the assay achieved a Z' factor of 0.74. We successfully screened the Library of Pharmacologically Active Compounds (LOPAC) for inhibitors of UHRF1_SRA binding to hemimethlylated DNA, and validated 6 hit compounds in a dose-response series. These include: aurintricarboxylic acid, NF449, 6-hydroxy-DL-DOPA, NF023, PPNDS and mitoxantrone. Additionally, we observed a decrease in methylation one week after treatment of DU145 cells with mitoxantrone at a concentration near its IC50 from the TR-FRET binding assay. This suggests that mitoxantrone, a known DNA topoisomerase II poison and general DNA intercalator, may interfere with methylation maintenance. In summary, we have established a novel method to identify small molecule inhibitors of the UHRF1-methylated DNA interface, and have shown that one hit compound, mitoxantrone, can perturb methylation maintenance near its IC50 for inhibition of SRA domain binding to methylated DNA.

Citation Format: David A. Walker, Nicolas Wyhs, Hugh Giovinazzo, Srinivasan Yegnasubramanian, William G. Nelson. Development of a high-throughput screening assay to identify UHRF1 inhibitors via time-resolved fluorescence resonance energy transfer (TR-FRET). [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 5390. doi:10.1158/1538-7445.AM2014-5390
Abstract 5399: A novel treatment for triple-negative breast cancer using intrinsic magnetized paclitaxel
Authors: Umemura Masanari , Makino Ayako , Sato Itaru , Feng Xianfeng , Oda Kayoko , Ohtake Makoto , Izuka Satoshi , Iwai Maki , Matsuo Kosuke , Eguchi Haruki , Ishikawa Yoshihiro
Published: May 2015

Background: We previously reported the identification of a novel nano-organic compound, EI236, an anti-cancer agent with intrinsic magnetic property. In addition to anti-cancer effect, its ferromagnetic property contributes to unique features.1) It can be attracted by a magnet. 2) It can be visualized by magnetic resonance imaging (MRI). Hereby, we have identified the key mechanism that contributes to magnetism by X-ray crystallographic analysis, and succeeded in generating a novel paclitaxel with intrinsic magnetism; this is a single paclitaxel compound, and is not a paclitaxel encapsulated in micelle with magnetic particles. Our aim is to examine its effect on triple negative-breast cancer (TNBC) cells. Material and Method: The magnetization of the magnetized paclitaxel was measured with a superconducting quantum interference device (SQUID) (Quantum Design MPMS7 system). Breast cancer cells, MDA-MB-453 (TNBC) and MCF7 (Non-TNBC), were obtained from RIKEN Bioresource center. Cell proliferation assay was performed using a commercially available kit, XTT Cell Proliferation Assay Kit. Apoptotic cells were stained with Annexin V, allophycocyanin conjugate and 7-amino-actinomycin D, and measured by fluorescence activated cell sorting (FACS), to evaluate early and late apoptosis. Cell cycle analysis was performed using The Cycletest™ Plus DNA Reagent Kit and assessed using FACS. Results: Plots of magnetization versus magnetic field revealed that the magnetized paclitaxel exhibits magnetic property in SQUID. Magnetized paclitaxel was easily attracted by a commercial bar magnet. Magnetized paclitaxel exhibits greater anti-cancer effect than original paclitaxel in TNBC and Non-TNBC cells in a dose-dependent manner. Magnetized paclitaxel induced apoptosis and G2/M arrest in cell cycle analysis in a dose-dependent manner, suggesting that magnetized paclitaxel retained the original anti-cancer property. In MRI T2 -weighted imaging, signal intensity was changed in a concentration-dependent manner with magnetized paclitaxel, but not with commercial available paclitaxel. Conclusion: These results suggested that various conventional anti-cancer drugs might be similarly magnetized, leading to novel drug development in future cancer chemotherapy.

Citation Format: Masanari Umemura, Ayako Makino, Itaru Sato, Xianfeng Feng, Kayoko Oda, Makoto Ohtake, Satoshi Izuka, Maki Iwai, Kosuke Matsuo, Haruki Eguchi, Yoshihiro Ishikawa. A novel treatment for triple-negative breast cancer using intrinsic magnetized 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 5399. doi:10.1158/1538-7445.AM2014-5399
Abstract 5519: Investigation of the cellular mechanisms of action of maximiscin, a novel natural product with selective cytotoxic activity against a triple negative breast cancer molecular subtype
Authors: Robles Andrew J. , Du Lin , King Jarrod B. , Cichewicz Robert H. , Mooberry Susan L.
Published: May 2015

Natural products have historically been an excellent resource for new pharmaceuticals, particularly anticancer agents. Fungi are a remarkable source of novel secondary metabolites that have never been evaluated for their therapeutic potential. However, a vast majority of fungal species found in diverse environments are inaccessible to research scientists. We recently isolated and identified the natural product maximiscin from a Tolypocladium sp. isolate obtained through a crowdsourcing initiative. This compound showed in vitro cytotoxic potency against multiple cancer cell lines in the NCI-60 in vitro screening panel and in vivo antitumor efficacy against a UACC-62 melanoma xenograft model. MDA-MB-468, a triple negative breast cancer (TNBC) cell line of the basal-like 1 (BL1) subtype, was found to be particularly sensitive to the cytotoxic effects of maximiscin compared to several other TNBC cell lines. This finding suggests maximiscin modulates a cellular signaling pathway that specifically drives the BL1 subtype of TNBC. In this study we investigated the cellular mechanisms of action of maximiscin using cell biological and biochemical approaches. The effects of maximiscin on cell cycle distribution were evaluated and maximiscin reduced the fraction of cells in the S and G2/M phases of the cell cycle, and began to induce apoptosis within 8 h. Intracellular signaling through multiple pathways was evaluated by immunoblotting, providing an indication of potential mechanisms of action. Maximiscin also circumvented P-glycoprotein-mediated multidrug resistance. These studies suggest maximiscin may have therapeutic potential for the treatment of BL1 TNBC, especially in cases where P-glycoprotein-mediated multidrug resistance has developed. Studies are ongoing to further characterize the cellular and antitumor effects of maximiscin in BL1 models.

Citation Format: Andrew J. Robles, Lin Du, Jarrod B. King, Robert H. Cichewicz, Susan L. Mooberry. Investigation of the cellular mechanisms of action of maximiscin, a novel natural product with selective cytotoxic activity against a triple negative breast cancer molecular subtype. [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 5519. doi:10.1158/1538-7445.AM2014-5519
Abstract 609: Targeting IGF-I receptor for hepatocellular carcinoma
Authors: Adachi Yasushi , Matsunaga Yasutaka , Yamamoto Hiroyuki , Nosho Katsuhiko , Suzuki Hiromu , Arimura Yoshiaki , Endo Takao , Kato Yasuo , Carbone David P. , Shinomura Yasuhisa
Published: May 2015

Aims; Hepatocellular carcinoma (HCC) is one of worse prognostic diseases. Insulin-like growth factor (IGF)-I receptor (IGF-IR) signaling is required for carcinogenicity and tumor development of many malignancies and is thought to be a next molecular target. We have previously shown successful therapy for gastrointestinal carcinomas, including colon, stomach, esophageal, pancreatic, and biliary tract cancers, using recombinant adenoviruses expressing dominant negative IGF-IR (IGF-IR/dn). In this study, we want to assess the roles of IGF axis on HCC and to develop potential targeted therapeutics for HCC.

Methods; We assessed mRNA expressions of IGFs and those receptors in HCC cell lines. We assessed the effect of IGF-IR blockade on signal-transduction, growth, apoptosis-induction, and migration. To block IGF signals, we used IGF-IR/dn, short hairpin RNA, monoclonal antibody, and tyrosin kinase inhibitor for IGF-IR.

Results; IGF-I messages expressed in 63%, IGF-II mRNA in 44%, and IGF-IR in 89% of HCC cell lines. All IGF-IR blockades suppressed significantly both in vitro cell growth and colony formation. Each anti-IGF-IR up-regulated both stressor- and chemotherapy-induced apoptosis. IGF-IR inhibitor down-regulated cell migration. IGF-IR targeting strategies effectively blocked ligands induced receptor activation and the downstream signals.

Conclusions; IGF axis might play a key role in tumor progression of human HCC. Every strategy to block IGF-IR shows a useful anticancer therapeutic for HCC, indicating that IGF-IR may be a candidate molecular target for HCC.

Citation Format: Yasushi Adachi, Yasutaka Matsunaga, Hiroyuki Yamamoto, Katsuhiko Nosho, Hiromu Suzuki, Yoshiaki Arimura, Takao Endo, Yasuo Kato, David P. Carbone, Yasuhisa Shinomura. Targeting IGF-I receptor for hepatocellular carcinoma. [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 609. doi:10.1158/1538-7445.AM2014-609
Abstract CT218: Wilms′ tumor gene 1 (WT1) peptide-based cancer vaccine combined with gemcitabine for patients with advanced pancreatic cancer
Authors: Koido Shigeo , Nishida Sumiyuki , Homma Sadamu , Takeda Yutaka , Komita Hideo , Morita Satoshi , Ito Toshinori , Morimoto Soyoko , Oka Yoshihiro , Yanagisawa Satoru , Toyama Yoichi , Ikegami Masahiro , Nagano Hiroaki , Ohkusa Toshifumi , Tajiri Hisao , Sugiyama Haruo
Published: May 2015

Wilms′ tumor gene 1 (WT1) protein is an attractive target for cancer immunotherapy. To investigate the feasibility of a combination therapy consisting of gemcitabine and WT1 peptide for patients with advanced pancreatic cancer, 32 HLA-A*24:02-positive patients were enrolled. HLA-A*24:02-restricted, modified 9-mer WT1 peptide (3 mg) emulsified with Montanide ISA51 adjuvant (WT1 vaccine) was injected intradermally biweekly. Moreover, gemcitabine (1,000 mg/m2) was administrated on days 1, 8, and 15 of a 28-day cycle. Median survival time and one-year survival rate were 8.1 months and 29%, respectively. The association between longer survival and positive delayed-type hypersensitivity (DTH) to WT1 peptide alone was statistically significant, and longer survivors featured a higher frequency of memory-phenotype WT1-specific cytotoxic T-lymphocytes (CTLs) before and during treatment. In conclusion, the combination therapy was well-tolerated and promising treatment for advanced pancreatic cancer. DTH-positivity to WT1 peptide alone and a higher frequency of memory-phenotype WT1-specific CTLs could be useful predictive markers for clinical efficacy.

Citation Format: Shigeo Koido, Sumiyuki Nishida, Sadamu Homma, Yutaka Takeda, Hideo Komita, Satoshi Morita, Toshinori Ito, Soyoko Morimoto, Yoshihiro Oka, Satoru Yanagisawa, Yoichi Toyama, Masahiro Ikegami, Hiroaki Nagano, Toshifumi Ohkusa, Hisao Tajiri, Haruo Sugiyama. Wilms′ tumor gene 1 (WT1) peptide-based cancer vaccine combined with gemcitabine for patients with advanced 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 CT218. doi:10.1158/1538-7445.AM2014-CT218
Abstract CT330: Adjusting for confounding effects of treatment crossover in a randomized phase 2 study of dabrafenib plus trametinib in BRAF V600+ metastatic melanoma
Authors: Latimer Nicholas , Amonkar Mayur , Stapelkamp Ceilidh , Sun Peng
Published: May 2015

In a randomized phase 2 study (BRF113220), dabrafenib (150 mg twice daily) plus trametinib (2 mg once daily) improved progression-free survival (HR=0.39; 95% CI, 0.25-0.62; P<.001) vs dabrafenib alone. 54 patients were randomized to each arm. At the most recent data cut (March 2013), median duration of follow-up was 24 months and 52% of patients had died. ITT analyses of overall survival (OS) are likely to underestimate the true benefit of treatment because patients in the control arm (83%) were able to cross over to combination treatment upon progression. ITT median OS, a secondary endpoint, was 23.8 vs 20.2 months (HR=0.73; 95% CI, 0.43-1.24). We adjusted for confounding due to this crossover using randomization-based crossover adjustment methods: the rank-preserving structural failure time model (RPSFTM) and iterative parameter estimation (IPE) algorithm. Different assumptions about the durability of treatment effect were tested: “treatment group” analyses allow for a treatment effect that continues beyond treatment discontinuation, while “on treatment_observed” analyses assumed that the treatment effect disappears upon discontinuation. Results are presented in the Table. Reestimation of median OS following adjustment with both RPSFTM and IPE (possible only for the treatment group analyses) suggested an OS gain of 9.1 months with combination treatment vs dabrafenib alone (compared with a 3.6-month gain under ITT assumptions). The RPSFTM estimation method performed marginally better than the IPE analyses and may provide more robust results. After adjusting for the effect of crossover, the reduction in HRs to 0.47-0.50 suggests that the ITT analysis underestimates the effect of dabrafenib plus trametinib on OS. Results are uncertain due to the limiting assumptions associated with the RPSFTM and IPE methodology, small trial size, low number of observed deaths, and low number of control group patients who did not cross over.

Crossover Adjustment MethodAssumptionOS HR (95% CI)None (ITT analysis unadjusted for crossover)0.73 (0.43-1.24)RPSFTMTreatment group0.47 (0.13-1.66)On treatment–observed analyses0.49 (0.15-1.63)IPETreatment group0.50 (0.16-1.54)On treatment–observed analyses0.47 (0.14-1.61)

Citation Format: Nicholas Latimer, Mayur Amonkar, Ceilidh Stapelkamp, Peng Sun. Adjusting for confounding effects of treatment crossover in a randomized phase 2 study of dabrafenib plus trametinib in BRAF V600+ metastatic melanoma. [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 CT330. doi:10.1158/1538-7445.AM2014-CT330
Abstract CT332: Biomarker analysis predicts antitumor activity of Ipilimumab therapy in patients with refractory metastatic melanoma
Authors: Chakravarti Nitin , Glitza Isabella C. , Trinh Van A. , Bassett Roland L. , Hwu Wen-Jen , Prieto Victor G.
Published: May 2015

Background: Ipilimumab (Ipi) improves survival of patients (pts) with advanced melanoma. It is unknown which biomarkers predict antitumor activity.

Methods: 81 pts with advanced melanoma who had failed to at least one prior systemic treatment were included in the Ipi Expanded Access Program (EAP) (Table 1). All pts received induction therapy with Ipi 10 mg/kg i.v. q 3 weeks x4 doses. At week 24, pts without progressive disease or toxicity received maintenance therapy with Ipi q 12 weeks. Pertinent biomarkers (CTLA-4, PD-1, PD-L1, pAKT, pS6, fas-l, Ki67, and BRAF V600E) were analyzed by immunohistochemistry on pre-treatment tumor specimens and results were correlated with clinical outcomes.

Results: Median progression free survival (PFS) and overall survival (OS) were 3.7 and 10.6 mos. Higher stages, ECOG (1 and 2), elevated LDH, and low absolute lymphocyte count (ALC) at baseline were associated with poor survival. Of the 71 pts with progression, 32 received alternative treatment and 39 did not; the OS from the time of disease progression was 17 and 2.6 mos, respectively. Increased CTLA-4 in tumor (p=0.003) and TILs (p=0.02) correlated with poor OS. pS6 expression in TILs (p=0.04), high pAkt in tumor (p=0.03) correlated with poor PFS. Low CTLA-4 (p=0.05) and pAkt (p=0.04) expression in tumor cells correlated with better outcome. High CTLA-4 expression in TILs associated with poor OS (p=0.003) in pts who received alternative treatment post-ipi.

Conclusions: High CTLA-4 expression in tumor & TILs as well as activated Akt pathway in tumor cells correlated with poor clinical outcome. High CTLA-4 expression in TILs correlated with poor survival in pts received alternative therapy after Ipi. Analyses of biomarkers predict antitumor activity of Ipi.

Table 1.Patient characteristicsVariableLevelNPercentSexMale4960%Female3240%StageIIIc22%M1a67%M1b1114%M1c6277%ECOG04353%12835%21012%Primary Melanoma SiteAcral Lentiginous45%Cutaneous6175%Mucosal79%Uveal22%Unknown primary79%Brain MetastasesNo4454%Before Treatment2328%During Treatment1012%After Treatment45%ResponseCR1114%PR22%SD22%PD6681%Post-Ipi RadiationNo1235%Yes2065%Post-Ipi SurgeryNo1547%Yes1753%Post-Ipi ImmunotherapyNo2991%Yes39%Post-Ipi Targeted TherapyNo1856%Yes1444%Post-Ipi ChemotherapyNo516%Yes2784%

Citation Format: Nitin Chakravarti, Isabella C. Glitza, Van A. Trinh, Roland L. Bassett, Wen-Jen Hwu, Victor G. Prieto. Biomarker analysis predicts antitumor activity of Ipilimumab therapy in patients with refractory metastatic melanoma. [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 CT332. doi:10.1158/1538-7445.AM2014-CT332
Abstract CT404: Phase I/II study of VAL-083 in patients with recurrent glioblastoma multiforme
Authors: Shih Kent C. , Patel Manish R. , Bacha Jeffrey , Brown Dennis M. , Garner William J. , Steino Anne , Schwartz Richard S. , Kanekal Sarath , Li Mike , Lopez Lorena M. , Burris Howard A.
Published: May 2015

Median survival for patients with recurrent glioblastoma multiforme (GBM) is less than 6 months. Front-line systemic therapy is temozolomide, but chemo-resistance due to O6-methylguanine-DNA-methyltransferase (MGMT) activity has been implicated in poor outcomes. VAL-083 is a structurally unique bi-functional DNA alkylating small molecule drug that crosses the blood-brain barrier and accumulates in brain tumor tissue. Previous human clinical studies by several investigators have suggested that VAL-083 has anti-tumor activity against a broad range of tumor types including GBM. In preclinical in vitro studies, VAL-083 demonstrated activity in a wide range of cancer cell lines, including pediatric and adult GBM cell lines and GBM cancer stem cells. Notably, due to crosslink formation on N7 of guanine, VAL-083 overcomes chemo-resistance to MGMT in vitro. In light of extensive safety data from several clinical trials and promising efficacy signals in CNS tumors, DelMar initiated a new clinical study to determine the safety, tolerability, pharmacokinetics, anti-tumor activity in patients with recurrent GBM. This is an open-label, single-arm Phase I/II dose-escalation study in patients with histologically-confirmed initial diagnosis of malignant GBM. The study utilizes a 3+3 dose-escalation design. Patients receive VAL-083 i.v. on days 1, 2, and 3 of a 21 day cycle. GBM patients have previously been treated with surgery and/or radiation, if appropriate, and must have failed both bevacizumab and temozolomide, unless contraindicated. In the ongoing trial, cohorts 1 through 4 (10 mg/m2) have completed the trial successfully with no drug-related serious adverse events (SAEs), and maximum tolerated dose (MTD) was not yet reached. Enrollment for Cohort 5 (20 mg/m2) has been initiated. Pharmacokinetic analyses show dose-dependent increase in exposure with a short 1-2 hour plasma half-life and a Cmax of <200 ng/ml at 10 mg/m2. Historical data suggests a long half-life in the cerebrospinal fluid (CSF) (>20 hours) with preferential accumulation to brain tumor tissue. MGMT status of patients and drug concentration in the CSF are being evaluated when possible. ClinicalTrials.gov Identifier: NCT01478178

Citation Format: Kent C. Shih, Manish R. Patel, Jeffrey Bacha, Dennis M. Brown, William J. Garner, Anne Steino, Richard S. Schwartz, Sarath Kanekal, Mike Li, Lorena M. Lopez, Howard A. Burris. Phase I/II study of VAL-083 in patients with recurrent glioblastoma multiforme. [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 CT404. doi:10.1158/1538-7445.AM2014-CT404
Abstract CT408: Phase I study of one mixed chelates copper(II) compound, Casiopeína CasIIIia with antitumor activity and its mechanism of action
Authors: Ruiz-Azuara Lena , Bastian Gerard , Bravo-Gómez Maria Elena , Cañas Roberto Carlos , Flores-Alamo Marcos , Fuentes Ines , Mejia Carmen , García-Ramos Juan Carlos , Serrano Alberto
Published: May 2015

Metal complexes have gained a growing interest as pharmaceuticals for their use as diagnostic agents or as chemotherapeutic drugs[1]. In our group some efforts have been done in the development of anticancer agents employing essential metals such as the family of copper (II) compounds known as Casiopeínas®, with a general formula [Cu (N-N) (X-Y) H2O] NO3 where N-N is a diimine (phen or bipy) and X-Y is a bidentate ligand (acac, salal, aminoacidate, peptide, benzimidazol).

These compounds have shown cytostatic, cytotoxic and antineoplastic activity in vitro and in vivo[2]. Also albumin/Casiopeinas interaction is discussed. Among those one have selected that is CasIII-ia as the most adequate to go into clinical trial Phase I.

Although the mechanism of action is still not completely elucidated, experimental evidence suggests the interaction of coordination compounds with DNA (nuclear or mitochondrial) and their components and the generation of reactive oxygen species (ROS) as the main action pathways. Induction of apoptosis has been proved to be the main death tumoral cell pathway. DNA cleavage capacity, compared with the 4, 4´-diimine analogs, then is suggested an intercalation process as the first step for the DNA damage. Also calculations have been done in order to modeling the interaction between Casiopeínas and DNA [3, 4].

The clinical protocol phase I of the dimethyl bpy derivative with acac Cas IIIia is presented and the selection criteria of the drug.

[1] S. H. van Rijt, P. J. Sadler, Drug Discov. Today, 14 (2009) 1089-1097.

[2] L. Ruiz-Azuara, M.E. Bravo-Gómez, Curr. Med. Chem., 17 (2010) 3606-3615.

[3] R. Galindo-Murillo, J. Hernández-Lima, M. González-Rendón, F. Cortés-Guzmán, L. Ruíz-Azuara, R. Moreno-Esparza Phys. Chem. Chem. Phys., 13 (2011) 14511-14516.

[4] R. Galindo-Murillo, L. Ruiz-Azuara, R. Moreno-Esparza, F. Cortés-Guzmán Phys. Chem. Chem. Phys., 14 (2012) 15539-15546.

Citation Format: Lena Ruiz-Azuara, Gerard Bastian, Maria Elena Bravo-Gómez, Roberto Carlos Cañas, Marcos Flores-Alamo, Ines Fuentes, Carmen Mejia, Juan Carlos García-Ramos, Alberto Serrano. Phase I study of one mixed chelates copper(II) compound, Casiopeína CasIIIia with antitumor activity and its mechanism of action. [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 CT408. doi:10.1158/1538-7445.AM2014-CT408
Abstract DDT01-04: SGN-CD19A: A novel Anti-CD19 antibody drug conjugate
Authors: Albertson Tina M. , Sandalic Lali , Zhao Baiteng , Kostic Ana , Law Che-Leung
Published: May 2015

CD19, a member of the immunoglobulin superfamily, is a B-cell specific marker that is found on B cells as early as the pro-B cell stage. CD19 is maintained upon malignant transformation and is expressed in the majority of patients with B-lineage acute lymphocytic leukemia (ALL) and non-Hodgkin lymphoma (NHL).

SGN CD19A is a novel antibody-drug conjugate (ADC) composed of a humanized anti-CD19 monoclonal antibody conjugated to the microtubule-disrupting agent monomethyl auristatin F (MMAF) via a maleimidocaproyl (mc) linker. Upon binding to CD19, SGN-CD19A internalizes and releases cys-mcMMAF, which binds to tubulin and induces G2/M arrest and apoptosis in the targeted cells. In mice bearing B-lineage ALL and lymphoma xenografts, SGN-CD19A induced dose-dependent tumor growth delay and extended survival.

Two first-in-human, phase 1, dose-escalation studies are ongoing to investigate the safety, tolerability, pharmacokinetics (PK), and antitumor activity of SGN CD19A; one trial in adult and pediatric patients with relapsed or refractory (R/R) B cell ALL or highly aggressive B-cell lymphoma (CT.gov NCT01786096) and the second trial in adult patients with R/R B-cell NHL (CT.gov NCT01786135). Both trials employ a modified continual reassessment method (CRM) to estimate the dose-toxicity relationship and to estimate the maximum tolerated dose. Exploratory analyses include correlation of target (CD19) expression, target saturation, and apoptotic cellular markers with clinical response and clearance of leukemic blasts; dose and PK of SGN-CD19A will also be correlated with target saturation and intracellular levels of cys-mcMMAF.

Early PK analyses indicate that plasma SGN-CD19A ADC generally increases with higher doses. Rapid clearance of ADC was observed at low doses in patients with leukemia, suggesting target-mediated drug disposition.

Dose escalation of SGN-CD19A is ongoing in both clinical trials. In preliminary safety analyses, SGN-CD19A was generally tolerable at doses examined to date. Encouraging antitumor activity has been observed, with multiple patients achieving complete remissions in both trials.

Citation Format: Tina M. Albertson, Lali Sandalic, Baiteng Zhao, Ana Kostic, Che-Leung Law. SGN-CD19A: A novel Anti-CD19 antibody drug conjugate. [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 DDT01-04. doi:10.1158/1538-7445.AM2014-DDT01-04
Abstract LB-150: Local maturation and inhibition of antitumor CD8 T-cell responses in a model of immunoevasive breast cancer
Authors: Boldajipour Bijan A. , Krummel Matthew F.
Published: May 2015

The purpose of this study is to investigate the cell interactions and mechanisms that limit anti-tumor CD8 T cell responses in tumor microenvironments.

We used cutting-edge multiphoton imaging of immunoevasive tumors to determine the behavior and anti-tumor responses of CD8 T cells. This was achieved by adoptively transferring T cells into a modified MMTV-PyMT breast tumor model (mouse mammary tumor virus promoter-driven, polyoma middle T antigen-induced luminal breast cancer). In this model, tumor cells expresses ovalbumin as tumor-antigen and mCherry as a fluorescent markers, thus allowing immunologic assessment and intravital imaging of T cell activity against tumor cells in and ex vivo. Furthermore, we were able to investigate the effect of antigen-presenting immune cells and immunomodulatory treatment regimens on anti-tumor T cell activity in and ex vivo.

In this work we found that newly arriving tumor-specific T cells undergo defined phases of maturation within the tumor microenvironment. First, after arriving to the tumor site, T cells engage in close interactions with antigen-presenting cells and exhibit reduced motility and rounded morphology. After this initial phase, T cells dramatically increase motility and survey the tumor microenvironment, however they do not form stable interactions with other cell types and further do not engage tumor cells. In contrary to other publications, swarming T cells do not express surface markers associated with exhaustion or anergy, and exhibit extensive in situ proliferation and ex vivo CTL activity. The capability of tumor CD8 T cells to kill targets ex vivo conflicts with the absence of tumor cell death in vivo. Additionally, individual immune cell populations isolated from tumors or tumor culture supernatants do not exhibit T cell suppressive capability when added as supplements to killing assays.

Our results define sequential fates for T cells entering tumor microenvironments, suggesting that tumor-infiltrating lymphocytes exist in distinct ‘states’ depending on the extent to which tumor antigen-presenting cells have matured them. Further, T cell suppression appears to be a local phenomenon and cannot be achieved by soluble factors or a specific cell type alone. This may suggest that combinations of therapies that individually target the different maturation states are required to achieve therapeutic effects.

Citation Format: Bijan A. Boldajipour, Matthew F. Krummel. Local maturation and inhibition of antitumor CD8 T-cell responses in a model of immunoevasive breast 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 LB-150. doi:10.1158/1538-7445.AM2014-LB-150
Abstract LB-206: Role of Olig2 phosphorylation in pediatric glioma formation
Authors: GRIVEAU Amelie , TIEN An-Chi , Rowitch David H.
Published: May 2015

Malignant gliomas are a leading cause of tumor-related mortality in children. Current therapies often have significant adverse effects on brain development and are largely ineffective against aggressive gliomas. The development of rational therapies has been limited by poor information regarding the cell of origin for pediatric glioma and the relevant underlying genetic alterations. Recent findings now reveal that mutations existing in pediatric glioma are different than those seen in adult gliomas. For example, BRAFV600E activating mutations are associated with deletion of the tumor suppressor gene, p16 (Ink4a-Arf), in about 15% of pediatric gliomas. We have generated a faithful genetic mouse model in which there is concomitant activation of BRAFV600E and deletion of Ink4a-Arf in oligodendrocyte progenitor cells (OPCs) leading to the generation of pediatric high-grade glioma. OPCs have been shown to be a potential cell of origin for glioma and expression of Olig2 is found in 100 percent of human pediatric and adult high-grade gliomas. Given that Olig2 is also required for normal myelin development, targeting all Olig2-expressing cells with small molecule inhibitors would likely have detrimental effects. We have recently found that Olig2 protein is present in a phosphorylated form that promotes proliferation of adult glioma cells. In this study, we demonstrate that phospho-Olig2 is expressed in tumorigenic oligodendrocyte progenitor cells and is required for pediatric glioma generation. In addition, we found that phospho-Olig2 and Olig2 interact with different proteins, some of which are feasible drug targets. This study gives insights into new therapies by understanding key protein interactions in normal oligodendrocyte development as well as in tumorigenic oligodendrocyte progenitor cells.

Citation Format: Amelie GRIVEAU, An-Chi TIEN, David H. Rowitch. Role of Olig2 phosphorylation in pediatric glioma formation. [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 LB-206. doi:10.1158/1538-7445.AM2014-LB-206
Abstract LB-261: Atopic allergic conditions and colorectal cancer risk in the Multiethnic Cohort
Authors: Tambe Neal A. , Stram Dan O. , Wilkens Lynne R. , Wan Peggy , Gilliland Frank D. , Henderson Brian E. , Marchand Loic L. , Haiman Christopher A.
Published: May 2015

Previous studies have provided suggestive evidence of an inverse association between atopic allergic conditions (AAC) and colorectal cancer (CRC) incidence and mortality in predominantly White populations. Here we examined the association between AAC and CRC among White, African American, Native Hawaiian, Japanese American, and Latino men and women in the Multiethnic Cohort (MEC). AAC exposure status was ascertained via self-reported physician diagnosis using a baseline 26-page validated questionnaire. The analysis included 4259 incident CRC cases and 1221 CRC-related deaths ascertained between 1993 and 2010. We also examined associations by ethnic group, location, and stage of cancer as well as potential effect modification by CRC risk factors. AAC was associated with a reduced risk of colorectal cancer incidence among both men and women (RR = 0.85, 95% CI 0.79, 0.91, p < 0.0001). The reduction in risk was noted in each population and was significant in Whites (RR = 0.84, 95% CI 0.71, 0.98, p = 0.03), African Americans (RR = 0.81, 95% CI 0.69, 0.95, p = 0.01), and Japanese Americans (RR = 0.85, 95% CI 0.75, 0.97, p = 0.01). Suggestive evidence of effect modification was noted by smoking (p = 0.07) and aspirin use (p = 0.08), with a greater protective effect observed among current smokers (RR = 0.72, 95% CI 0.58, 0.89, p = 0.002) and never users of aspirin (RR = 0.81, 95% CI 0.73, 0.89, p < 0.0001). Individuals with AAC also had a 19% reduction in CRC-related mortality (P = 0.004). Colonoscopy screening is unlikely to explain these inverse associations, as individuals with AAC were more likely to be screened than those without AAC (p < 0.003 for all ethnic groups). These findings provide support for an association between immune pathology and colorectal cancer, potentially through modulation of the immune system's IgE-mediated pathways systemically and within the gut.

Citation Format: Neal A. Tambe, Dan O. Stram, Lynne R. Wilkens, Peggy Wan, Frank D. Gilliland, Brian E. Henderson, Loic Le Marchand, Christopher A. Haiman. Atopic allergic conditions and colorectal cancer risk in the Multiethnic Cohort. [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 LB-261. doi:10.1158/1538-7445.AM2014-LB-261