bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1147: Probing the roles of SUMOylation in cancer cell biology using a selective SUMO activating enzyme inhibitor
Authors: Pulukuri Sai M , He Xingyue , Grossman Stephen , Riceberg Jessica , Koenig Erik , Soucy Teresa , Song Keli , Kreshock Anna , England Dylan , Mizutani Hirotake , Dick Larry , Brownell James , Newcomb John , Langston Steve , Lightcap Eric , Galvin Katherine
Published: July 2017

SUMOylation has been implicated in many cellular processes that are important for cancer cell survival, including cell cycle, chromosome structure and segregation, nuclear and subnuclear organization, transcription and DNA damage repair. However, a potent and selective inhibitor to target the SUMO pathway has been lacking. The SUMO-activating enzyme (SAE) is an essential enzyme in the pathway that initiates the SUMOylation process. Here we report the identification of the first mechanism-based SAE inhibitors with nanomolar potency in cellular assays. These inhibitors selectively block SAE enzyme activity and total SUMOylation in cells, which leads to reduced cancer cell proliferation. Moreover, SAE inhibition resulted in disruption of PML nuclear bodies and redistribution of DAXX. In vivo administration of SAE inhibitor into tumor bearing mice results in modulation of several biomarkers including a significant reduction in SUMO-conjugates and E2 Ubc9 thioesters, demonstrating SUMO pathway inhibition. Our results demonstrate the feasibility of inhibiting the SUMO pathway with small molecule inhibitors and provide tools to study the SUMO biology in cancer.

Citation Format: Sai M Pulukuri, Xingyue He, Stephen Grossman, Jessica Riceberg, Erik Koenig, Teresa Soucy, Keli Song, Anna Kreshock, Dylan England, Hirotake Mizutani, Larry Dick, James Brownell, John Newcomb, Steve Langston, Eric Lightcap, Katherine Galvin. Probing the roles of SUMOylation in cancer cell biology using a selective SUMO activating enzyme inhibitor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1147. doi:10.1158/1538-7445.AM2017-1147
Abstract 115: Flubendazole targets cancer stem-like properties and the HER2/Akt signaling pathway in HER2-positive breast cancer with trastuzumab resistance
Authors: Sung Daeil , Cho Youngkwan , Oh Eunhye , Cho Tae-Min , Kim Yoon-Jae , Kim Ji Young , Seo Jae hong
Published: July 2017

Background and Propose: HER2-positive breast cancers are associated with an aggressive phenotype and trastuzumab resistance leading to poor clinical outcomes. Flubendazole (FLU) is a potent anthelmintic agent that inhibits microtubule polymerization, and also exhibits anticancer activity in several cancer types. The objective of the present study was to investigate the effect of FLU on apoptosis, HER2/Akt signaling pathway, cancer stem cell (CSC)-like properties and trastuzumab-resistance in HER2-positive breast cancer cells. Methods: FLU-induced antitumor effects were characterized by MTS assay, Sub-G1 and G2/M-phase arrest population, Annexin-V analysis, ALDH1 activity assay, Western blotting and immunofluorescence. For in vivo study, trastuzumab-resistant JIMT1 cells were injected into the mammary fat pads of BALB/c nude mice. We then determined the tumor growth and CSC-like properties. Results and conclusion: FLU significantly reduced cell viability and induced apoptosis with accompanied by activation of caspase-3, -7, and -8, as well as PARP cleavage in HER2-positive cell lines (MDA-MB-453, SKBR3 and BT474). FLU-induced apoptosis is associated with significant decreases in expression levels of HER2, p-HER2 (Tyr1221/1222), HER3, and p-HER3 (Tyr1289). FLU treatment also resulted in G2/M-phase arrest of the cell cycle, as evidenced by a marked downregulation of phospho-Histone H3 expression. These results were closely related to the decline of CSC-like properties, coinciding with the suppression of ALDH1 activity. These effects were also observed in trastuzumab-resistant JIMT-1 cells, and significant reductions in tumor growth and ALDH1A1 downregulation were observed in xenograft models together with downregulation of HER2 expression in vivo. Our findings suggest that FLU may be potentially effective for the treatment of HER2-positive breast cancer with trastuzumab resistance.

Note: This abstract was not presented at the meeting.

Citation Format: Daeil Sung, Youngkwan Cho, Eunhye Oh, Tae-Min Cho, Yoon-Jae Kim, Ji Young Kim, Jae hong Seo. Flubendazole targets cancer stem-like properties and the HER2/Akt signaling pathway in HER2-positive breast cancer with trastuzumab resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 115. doi:10.1158/1538-7445.AM2017-115
Abstract 1157: Screening and identification of new generation glucose transporter inhibitors as anticancer therapeutics
Authors: Qian Yanrong , Shriwas Pratik , Wang Xuan , Roberts Dennis , Kessler Emma , Bergmeier Stephen , Chen Xiaozhuo
Published: July 2017

Cancer is a top killer second only to cardiovascular diseases. Cellular metabolic reprogramming and altered energetics are key features of cancer. Opportunistic uptake of extracellular nutrients has recently been named as an emerging hallmark of cancer metabolism. Glucose is a key nutrient taken in by cancer cells for satisfying their high energy and biosynthesis needs. Drastic upregulation of glucose uptake and glycolytic activity, a phenomenon called the Warburg effect, is prevalent in cancers. Cancer cells overexpress specific glucose transporters (Gluts), particularly Glut1, on cell membranes for taking in glucose via facilitated diffusion. Considering the addiction of cancer cells for glucose and their sensitivity and vulnerability to changes in glucose supply, Gluts, especially Glut1, have been recognized as an attractive anticancer target. We previously reported the identification of first generation lead Glut1 inhibitor WZB117 (1). WZB117 treatment of human nonsmall cell lung cancer (NSCLC) A549 cells resulted in inhibition of glucose uptake, glycolysis and induction of apoptosis and necrosis. In addition, WZB-117 reduced tumor growth by ~70% in an A549 tumor mouse model. WZB117 inhibit cell growth of 7 cancer cell lines in 4 cancer types with IC50 values lower than 10 μM. Through a structure activity relationship (SAR) study of WZB117, a second generation lead, DRB18, has been identified. DRB-18 is much more stable and 5 to 10 times more potent than WZB117 in inhibiting growth and proliferation in about 90% of the 60 different cancer cell lines in 9 major cancer types as revealed in NCI-60 screenings. And its IC50s in many cancer cell lines are in the high nM range. In this study, we also report new compounds screening results for the objective of identifying novel compounds with higher glucose transport-inhibitory and cancer cell growth-inhibitory activities using nonsmall cell lung cancer cell lines as models. References: 1. Liu, Y., Cao, Y., Zhang, W., Bergmeier, S., Qian, Y., Akbar, H., Colvin, R., Ding, J., Tong, L., Wu, S. and Hines, J., 2012. A small-molecule inhibitor of glucose transporter 1 downregulates glycolysis, induces cell-cycle arrest, and inhibits cancer cell growth in vitro and in vivo. Molecular cancer therapeutics, 11(8):1672-1682.

Citation Format: Yanrong Qian, Pratik Shriwas, Xuan Wang, Dennis Roberts, Emma Kessler, Stephen Bergmeier, Xiaozhuo Chen. Screening and identification of new generation glucose transporter inhibitors as anticancer therapeutics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1157. doi:10.1158/1538-7445.AM2017-1157
Abstract 1182: A novel curcumin derivative inhibits active ras and its downstream pathway in pancreatic cancer
Authors: Mallangada Naveen , Mackenzie Gerardo G.
Published: July 2017

Pancreatic Cancer’s (PC’s) 5-year survival rate of only 6.7% indicate the need to improve treatment modalities. Despite decades of research, current chemotherapy and radiation therapy regimens offer minimal or no help. It is critical to develop new agents for the effective management of PC. Given that Kras mutations initiate and maintain PC, inhibition of this pathway is widely considered a therapeutic target of exceptional importance. Our laboratory is currently exploring the chemotherapeutic efficacy of a novel Chemically-Modified Curcumin (CMC2.24) as a potential chemotherapeutic agent for PC. Preliminary studies have shown that CMC2.24 has higher bioavailability than curcumin, as shown in pharmacokinetic studies in rats, and inhibits PC growth in vitro and in vivo. However, the exact mechanism on how CMC2.24 reduces cell growth remains unidentified. The objective of this work was to determine the mechanism of CMC2.24 in PC. Using human PC MIA PaCa-2 and Panc-1 cell lines and pancreatic acinar explants from Kras mutant mice, we explored the effects of CMC2.24 on Ras activation, ERK phosphorylation, mitochondrial reactive oxygen species, mitochondrial ATP production, and intrinsic apoptosis. In human PC MIA PaCa-2 cells CMC2.24 inhibited Ras activation by 90% (p<0.05). This was confirmed in primary acinar explants isolated from Kras mutant mice in which CMC treatment reduced Ras activation by 70%, compared to control. Furthermore, CMC 2.24 treatment reduced the phosphorylation MEK, ERK and c-Raf, down-stream pathway of Ras, both in vitro and in vivo. The effect of CMC2.24 on ERK phosphorylation was confirmed by Immunofluorescence, showing a significant decrease in ERK phosphorylation (47.8%; p<0.05). Moreover, CMC2.24 treatment enhanced the levels of superoxide anion in mitochondria by 200% (p<0.02), decreased ATP levels in a concentration-dependent manner (p<0.05), and induced intrinsic apoptosis, as shown by the increase in caspase 9 and Parp cleavage downstream of cytochrome C cytosolic release. In conclusion, our results indicate that the Ras pathway is a key molecular target for CMC2.24 and that CMC induces apoptosis in PC cells through the intrinsic pathway.

This research was supported by the Stony Brook Cancer Center and a URECA Summer Grant.

Citation Format: Naveen Mallangada, Gerardo G. Mackenzie. A novel curcumin derivative inhibits active ras and its downstream pathway in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1182. doi:10.1158/1538-7445.AM2017-1182
Abstract 1190: Comprehensive high-throughput screen for combination therapies to block acquired resistance to targeted drugs
Authors: Haviv Izhak
Published: July 2017

Background - Genetic and epigenetic alterations provide the selective advantage for cancer cell acquired drug resistant (ADR). A bottleneck in implementing drug combination is the challenging task of identifying combinations with acceptable risk benefit ratio. High throughput functional genomic (RNAi) screen is an ideal tool to comprehensively identify double and triple inhibitor combinations to prevent ADR. Unfortunately, most anticancer agents are antagonists (antagonists operate in the same direction as the RNAi), therefore RNAi hits with possible clinical value are the ones that disappear during the screen.

Materials and Methods - We use lentiviral-borne shRNA libraries (mainly kinome collection) to transduce the different RNAis into patient-derived cancer cells. The resulting mixed cancer cell population (each lost one gene) is subject to negative screen, to identify mediators of ADR. This process was performed with approved cancer drugs, such as erlotinib, olaparib, pazopanib, etc. We developed a proprietary unique way to identify the RNAi clones that disappear/extinct, specifically in the presence of the investigated drug. RNAi clones are tracked using next generation sequencing, and quantified in treated and untreated cancer cell cultures or xenografts. The resulting drug combinations identified through this screen are tested directly on a platform of patient-derived xenografts.

Results - We identified known boosters of the EGFR activity, such as DYRK1 (maintains level of EGFR protein), IKBKE (downstream survival mediator), which when reduced by their RNAi, enhance the response to erlotinib. Consequently, known inhibitors of JAK1, CDK4/6, and PI3K combined to egfr inhibitors generated more durable remissions in egfr positive cells. Remarkably, even tumors without egfr mutations were sensitive to the combinations. A similar discovery of novel combinations were found for olaparib and pazopanib. Mouse harboring patient derived xenografts tolerated combination therapy fairly well.

Conclusion - Our findings demonstrate the value of clinically testing the efficacy of drug combinations empirically designed to block ADR. Animal model support our findings with acceptable toxicity.

Citation Format: Izhak Haviv. Comprehensive high-throughput screen for combination therapies to block acquired resistance to targeted drugs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1190. doi:10.1158/1538-7445.AM2017-1190
Abstract 1209: FGFR inhibition re-sensitizes BRAF/MEK dual resistant cells to the BRAF/MEK inhibitor combination
Authors: Wang Victoria E. , Settleman Jeffrey , McCormic Frank
Published: July 2017

The BRAF-MEK pathway is frequently mutated in human melanoma. Inhibitors of these kinases have proven to prolong survival in melanoma patients. However, clinical benefit is relatively short-lived due to acquired drug resistance. To explore mechanisms of resistance in melanoma cells treated with dual BRAF and MEK inhibitors, A375, a cell line harboring the BRAF V600E mutation, was treated with escalating doses of vemurafenib and cobimetinib to generate a cellular population resistant to combination therapy. These cells display a reversible resistance phenotype. Upon initial treatment of the drug combination, upregulation of pSTAT3 is observed but the cells lose dependency on the pSTAT3 pathway upon prolonged drug treatment. A pharmacological synthetic lethal screen was performed and the FGFR inhibitor class re-sensitizes the cells to BRAF and MEK inhibition through inhibitions of pERK. The dual resistant cells also displayed hyperactivation of the PI3K-AKT pathway. Interrogation of patient samples confirmed these findings. These findings provides a biological rationale for a polytherapy strategy using a FGFR inhibitor in patients with resistance to dual BRAF and MEK inhibition.

Citation Format: Victoria E. Wang, Jeffrey Settleman, Frank McCormic. FGFR inhibition re-sensitizes BRAF/MEK dual resistant cells to the BRAF/MEK inhibitor combination [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1209. doi:10.1158/1538-7445.AM2017-1209
Abstract 1233: WNT inhibitor ICG-001 prevents visceral metastatic triple negative breast cancer in a chemo-resistant patient derived xenograft -PDX-model
Authors: Fatima Iram , Ayachi Ikbale El , Zacheaus Chidi , Gray Joseph Kerby , Krutilina Raya , Seagroves Tiffany N. , Krum Susan A. , Miranda-Carboni Gustavo A.
Published: July 2017

Triple negative breast cancer (TNBC) is characterized by absence of the estrogen receptor (ER) and progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) amplification. Such cancers are highly aggressive and frequently metastasize to the lung and brain. Unlike other breast cancer subtypes such as, ER+, PR+ and/or HER2+, TNBCs have no specific targeted-therapeutics; therefore, studies should be directed for development of targeted therapies to treat this condition. Recent studies indicate that Wnt/β-catenin signaling is particularly activated in TNBC and is associated with reduced overall survival in all patients. Therefore, pharmacological targeting of Wnt signaling pathway constitutes an ideal approach for treating TNBC and various Wnt inhibitors are currently in use in clinical trials, such as; ICG-001 in metastatic colon cancer. The inhibitory effects of ICG-001 have not been tested in chemoresistant TNBC PDX tumors. Herein, we report for the first time that ICG-001 compound selectively inhibited the growth of several European American (EA) and African American (AA) triple negative breast cancer subtypes MSL and BSL-1 cell lines both in vitro and in vivo models. To further investigate the precise mechanisms of action in the regulation of Wnt/β-catenin signaling by ICG-001, we performed Western blot analysis, apoptosis assays, cell cycle assays and quantitative real-time reverse transcriptase- polymerase chain reactions in human triple negative breast cancer cells. This compound significantly interfered with Wnt/β-catenin signaling, and its inhibition led to downregulation of important downstream targets such as Axin2, HMGA2, PCNA, c-myc and Cyclin D1, which in turn led to inhibition of proliferation, cell cycle progression and metastasis confirming our previous results too. In addition ICG-001 inhibited the invasion and motility of tumor cells and showed inhibition and prevention of visceral metastatic PDX tumors from both chemoresistant EA and AA women. These results indicate that the Wnt inhibitor ICG-001 could constitute a powerful new chemotherapeutic agent against triple negative breast cancer.

Citation Format: Iram Fatima, Ikbale El Ayachi, Chidi Zacheaus, Joseph Kerby Gray, Raya Krutilina, Tiffany N. Seagroves, Susan A. Krum, Gustavo A. Miranda-Carboni. WNT inhibitor ICG-001 prevents visceral metastatic triple negative breast cancer in a chemo-resistant patient derived xenograft -PDX-model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1233. doi:10.1158/1538-7445.AM2017-1233
Abstract 1241: A preclinical study of the combined treatment of arginase and canavanine in pancreatic cancer
Authors: Kwong Tsz Tung , Wong Chi Hang , Loong Herbert Ho Fung , Chan Stephen Lam
Published: July 2017

Background: Arginine deprivation is a novel approach for cancer treatment as some cancers were found to be arginine auxotrophic. PEG-BCT-100, a pegylated form of recombinant human arginase, was used to investigate the effect of arginine deprivation on pancreatic cell lines. On the other hand, Canavanine (CAV), a natural toxic analog of arginine isolated from leguminous plant, has also demonstrated its ability in inhibiting cancer cell proliferation. It alters the protein structure and causes critical metabolic defects in the cells by replacing arginine into newly synthesized proteins during translation. In this study, we have also tested the efficacy of combined arginase and CAV treatment on pancreatic cell lines.

Methods: Pancreatic cell lines (MIA PaCa-2, CFPAC-1) and normal fibroblast cell line (WI-38), were either incubated in normal or arginine free condition. The corresponding IC50 of CAV was determined by cell viability assay. To evaluate the responsiveness to the combined treatment, cells were monitored by live cell imaging under the conditions of PEG-BCT-100 treatment alone (0.3IU/ml) or in combination with CAV (10μM or 50μM). The apoptotic effect of the combined treatment was examined by Western blotting and Annexin V assay.

Results: Either arginase or CAV treatment could inhibit pancreatic cancer cell growth. The cells treated with CAV in arginine free condition demonstrated an inhibition of cell proliferation and its corresponding IC50 was significantly lowered from millimolar to micromolar concentration when arginine was withdrawn. Strong synergism of CAV and PEG-BCT-100 was observed after 48-hour treatment with enhanced apoptosis in pancreatic cell lines, but not in normal fibroblast cells. Both early and late apoptosis were observed in co-treated pancreatic cancer cells as indicated by Annexin V assay and the detection of polyADP ribosyl polymerase (PARP) in Western blot.

Conclusion: Arginine deprivation by PEG-BCT-100 combined with CAV treatment showed a strong synergetic effect on inducing cell death in pancreatic cell lines, suggesting that the supplement of CAV could facilitate the treatment outcome of arginine deprivation by arginase. This combined treatment may serve as a treatment strategy for pancreatic cancer.

Citation Format: Tsz Tung Kwong, Chi Hang Wong, Herbert Ho Fung Loong, Stephen Lam Chan. A preclinical study of the combined treatment of arginase and canavanine in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1241. doi:10.1158/1538-7445.AM2017-1241
Abstract 1248: Adult calorie restriction reduces weight gain and mammary tumor incidence in obesity prone MMTV-TFG α mice -
Authors: Grossmann Michael E. , Mizuno Nancy K. , Yang Da-Qing , Liao D. Joshua , Cleary Margot P.
Published: July 2017

Obesity is associated with increased risk of postmenopausal breast cancer resulting in recommendations for weight gain prevention and/or weight loss. However, there is little experimental data indicating if calorie restriction (CR) initiated in adulthood impacts mammary tumorigenesis. We have used female MMTV-TGF-α mice which develop mammary tumors in the 2nd yr of life as a model of postmenopausal breast cancer. From 10-30 weeks of age (woa) mice were fed a diet with 33% fat calories. At 30 woa based on body weight mice were classified as Obesity-Prone (Ob-Pr), Overweight (Ow) or Obesity-Resistant (Ob-R). Half in each weight category were then fed a 25% CR diet with reduced carbohydrate content (43% fat calories). The remaining mice continued on ad libitum (AL) feeding of the 33% fat diet. This resulted in six experimental groups (n = 26-31/group). Food intakes were monitored and body weights and mammary tumor status were determined weekly. Mice were sacrificed either at 90 woa or earlier if tumor size dictated. Mice in the CR-Ob-Pr group lost an average of 1.12 grams while mice in the AL-Ob-Pr group gained 16.03 grams from 30 woa until euthanasia. CR-Ob-Pr mice had a reduced palpable tumor incidence as compared to AL-Ob-Pr mice prior to euthanasia (12.90% versus 61.54% P = 0.0002). Indolent tumors were discovered at euthanasia with the CR-Ob-Pr mice having a larger number than AL-Ob-Pr mice resulting in final tumor positive mice in these groups of 54.84% vs 65.38%, respectively. Interestingly, the Ob-R mice had final tumor incidence of 38.71% and 73.08% for the CR-Ob-R vs the AL-Ob-R (P<0.01). Average tumor weight was highest in the AL-Ob-Pr group with CR resulting in a significantly lower tumor weight (P< 0.01). The AL-Ob-Pr group had the shortest average survival time of 82.96 woa while CR of the Ob-Pr mice resulted in increased survival to 89.77 woa (P < 0.001). A comparison of the survival curves of these two groups showed that CR resulted in significantly increased survival (P < 0.0001). Results of this study indicate that initiation of moderate 25% CR during adulthood reduces weight gain, mammary tumor incidence, and tumor weight as well as increases lifespan even when a moderately high fat diet was consumed. This information supports further efforts to be made to prevent weight gain as women approach menopause to delay or prevent breast cancer development. Support: NIH-CA157012, The Hormel Foundation and Paint the Town Pink.

Citation Format: Michael E. Grossmann, Nancy K. Mizuno, Da-Qing Yang, D. Joshua Liao, Margot P. Cleary. Adult calorie restriction reduces weight gain and mammary tumor incidence in obesity prone MMTV-TFG α mice - [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1248. doi:10.1158/1538-7445.AM2017-1248
Abstract 1263: Andrographolide inhibits prostate cancer by suppressing cytokine involved in promoting epithelial to mesenchymal transition
Authors: Mir Hina , Kapur Neeraj , Sonpavde Guru , Singh Shailesh
Published: July 2017

Mesenchymal phenotypes are responsible for cancer progression and poor therapeutic response. Hence, approaches targeting mesenchymal phenotype or inhibiting epithelial to mesenchymal transition will significantly prevent cancer progression or improve therapeutic outcome. Using in vitro models and molecular approaches we have shown that Andrographolide (AG), a compound isolated from Andrographis paniculata, shows promising effect on mesenchymal cells. It disposes PCa cells towards epithelial phenotype by modulating E-cadherin, ZEB-1, SNAIL and TWIST. In addition to these, significant inhibition in IL-6, which is known to be a poor prognostic maker, was observed in PCa cells treated with AG compared to untreated cells. This reduction in IL-6 could be due to AG induced suppression of SOCS (Suppressor of cytokine signaling) molecules. Therefore, our data underscores the ability of AG to impede cancer growth by impacting EMT and rationalizes its application as a potent therapeutic agent.

Citation Format: Hina Mir, Neeraj Kapur, Guru Sonpavde, Shailesh Singh. Andrographolide inhibits prostate cancer by suppressing cytokine involved in promoting epithelial to mesenchymal transition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1263. doi:10.1158/1538-7445.AM2017-1263
Abstract 1267: Emodin exerts its anticancer effect on colon cancer cells by inhibiting proliferation and inducing apoptosis
Authors: Saunders Ian T. , Mir Hina , Kapur Neeraj , Singh Shailesh
Published: July 2017

Despite state of the art diagnostic and therapeutic options offered in the clinics, colon cancer is still a major health care concern. Therapeutic outcome is often compromised due to late diagnosis and development of resistance against treatment options used to treat advanced disease. Hence, the need to explore new avenue to treat/prevent colon cancer more effectively still exist. In this study we have investigated potential impact of Emodin, an anthraquinone derived from plant (Rheum rhabarbarum) on colon cancer cell proliferation, survival and apoptosis. Our data show that emodin reduces the cell viability and inhibits proliferation of human colon cancer cell (COLO-201 and DLD-1) in a time-dependent manner. Furthermore, FACS analysis show higher percentage of apoptotic cells with Emodin treatment compared to untreated cells. Interestingly, pro-inflammatory and anti-inflammatory cytokines produced by colon cancer cells were modulated with Emodin treatment. Cancer cells often use these cytokines to achieve their goal to progress and escape the treatment. Our data implies Emodin could be a promising agent that could be used for chemoprevention as well as in therapeutic combination to improve efficacy of conventional therapy.

Citation Format: Ian T. Saunders, Hina Mir, Neeraj Kapur, Shailesh Singh. Emodin exerts its anticancer effect on colon cancer cells by inhibiting proliferation and inducing apoptosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1267. doi:10.1158/1538-7445.AM2017-1267
Abstract 1282: Analysis of missense variants in BRCA1 BRCT domains
Authors: Pietro Giuliano Di , Fernandes Vanessa Camara , Amankwah Kwabena , Shields Cara , Golubeva Volha A. , Harro Carly , Carvalho Marcelo Alex , Monteiro Alvaro N.
Published: July 2017

Currently, only ~¼ (133/600) of documented single nucleotide variation leading to missense changes in the BRCA1 gene have been characterized as pathogenic or non-pathogenic (IARC Classes 4 & 5 and 1 & 2, respectively) posing a challenge for carriers of these variants. A detailed understanding of structure-function relationships is critical for the assessment of these variants in the interpretation of data from functional assay and for the development of functional impact predictors. Thus, we have systematically probed the C-terminus of BRCA1 including the N-terminal border of the BRCT domains (M1650, S1651), the BRCT linker region (G1738, D1739, V1740, G1743, H1746, G1748, P1749, R1753, and D1757), and the alpha 1 and alpha’1 helices in BRCT[N] and [C], respectively. We assessed the functional impact of 47 missense variants using a validated transcriptional assay by a protein fusion of the GAL4 DNA-binding domain to BRCA1 C-terminus (aa 1,396-1,863) in HEK 293FT cells. Analysis of functional data redefined the BRCT border (previously thought to be at S1651) at residue M1650. Overall, mutations in the BRCT linker had severe impact on the function. Mutations in the alpha1 and alpha’1 helices also tended to display impaired function despite the lack of functional impact of 16 out of the 17 variants previously analyzed. In summary, our study provides increased granularity in our understanding of BRCA1 regions less tolerant to changes and therefore more likely to contain pathogenic variants. This knowledge will be critical for the assignment of different prior probabilities of pathogenicity given the variant location, improving risk assessment and clinical management of carriers of BRCA1 missense alleles

Note: This abstract was not presented at the meeting.

Citation Format: Giuliano Di Pietro, Vanessa Camara Fernandes, Kwabena Amankwah, Cara Shields, Volha A. Golubeva, Carly Harro, Marcelo Alex Carvalho, Alvaro N. Monteiro. Analysis of missense variants in BRCA1 BRCT domains [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1282. doi:10.1158/1538-7445.AM2017-1282
Abstract 1283: Mutations in the Duffy Antigen Receptor for Chemokines (DARC/ACKR1) gene result in population-private variants
Authors: Martini Rachel , Jenkins Brittany , Davis Melissa
Published: July 2017

By studying the breast cancer microenvironment, we can specifically address novel aspects of cancer development and progression at the molecular level. Chemokines are small signaling molecules that are an important component of this microenvironment, as they shape cellular composition of this space through recruitment of immune cells, among other functions. DARC, the Duffy Antigen Receptor for Chemokines, is an atypical chemokine receptor (ACKR1) that can bind different classes of chemokines. It is a non-signaling receptor that mainly acts as a regulator of chemokine homeostasis by removing them from circulation to recruit appropriate immune cell types. The purpose of this study is to determine how population-private variants of the DARC/ACKR1 gene across global population render distinct function and define aspects of human diversity in immune responses. A well-studied example of this would be the Duffy-null allele. This allele carries a mutation that is population-specific to recent decedents of Sub-Saharan Africans, and removes expression of DARC on red blood cells. Loss of expression on red blood cells presumably causes these “Duffy-Null” individuals to lose the ability to sequester chemokines, therefore losing homeostatic balance of these molecules in circulation. To identify additional mutations, we completed in silico analysis of minor allele frequencies across the gene in the 1000 Genomes Project data, and identified several other putative functional mutations that likely result in additional gene variants, specific to geographic ancestry groups and likely under similar selection as the Duffy null allele. We have prioritized our initial functional study to include mutations in the regulatory regions of the gene, as these mutations can change how and where the gene is expressed. Future work is focused on investigating these functional variants in situ, using human tissue and human breast cell lines, by isolating and re-engineering ancestry-specific variants in these cell lines. By identifying and determining functional outcomes of these population-specific mutation, we can observe differences in immune responses across these global populations, further defining the role that DARC/ACKR1 plays in the breast tumor microenvironment.

Citation Format: Rachel Martini, Brittany Jenkins, Melissa Davis. Mutations in the Duffy Antigen Receptor for Chemokines (DARC/ACKR1) gene result in population-private variants [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1283. doi:10.1158/1538-7445.AM2017-1283
Abstract 1285: Analysis of esophageal cancer SNPs and gene expression to predict ESCC risk
Authors: Ding Lilly A. , Ding Molly P. , Song Xin , Wang Lidong , Wang Liang
Published: July 2017

Esophageal cancer is one of most common cancers worldwide and its incidence has increased rapidly over the past decades. The majority of esophageal cancers is the esophageal squamous cell carcinoma (ESCC), which is particularly prevalent in Asian and South African areas. Most cases of ESCC are diagnosed at advanced stages, with an overall 5-year survival rate of 10–20%. Currently, genome-wide association studies (GWAS) are used to identify high risk individuals for the development of ESCC. Many SNPs have been identified to associate with a risk of ESCC. However, the effect size attributable to individual SNPs was typically modest, suggesting that genetic variants may only account for a very small amount of genetic risk and heritability of ESCC. The combined effect of multi genetic variants with altered mRNA expression play critical roles in ESCC. In this study, we used several existing bioinformatics databases (HaploReg, GTEx Portal and cBio Portal). We first selected 450 potential ESCC risk SNPs from HaploReg and then defined which SNPs caused gene expression alterations in the esophagus through expression quantitative trait loci (eQTL). Nine SNPs in eight genes (MUC1, ALS2CR12, ADHB1, TMEM173, XBP1, PLCE1, HEATR3 and SMG6) were identified as having altered mRNA expression. The altered mRNA expression of these seventeen genes are: MUC1, THBS3, GBAP1, EFNA1, SCAMP3, CASP8, CASP10, ALS2CR12, STRADB, ADH4, TMEM173, CYCTM1, DNAJC18, DNAJC20, NOC3L, HEATR3 and SRR. These altered gene expressions in the mucosal layer of the esophagus metabolize enzymes, repair DNA, and prevent cell death and inflammation, all of which may indicate that the existing altered esophageal mRNA levels plus the predisposition of normal esophageal epithelial cells can lead to harmful environmental carcinogens, thus attributing to ESCC development. In order to validate these gene alterations related to ESCC SNPs in screening and early detection of high risk individuals, DNA genotyping of this group of SNPs with a prospective cohort (n=5451) in a high risk area of ESCC will be tested in future studies.

Citation Format: Lilly A. Ding, Molly P. Ding, Xin Song, Lidong Wang, Liang Wang. Analysis of esophageal cancer SNPs and gene expression to predict ESCC risk [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1285. doi:10.1158/1538-7445.AM2017-1285
Abstract 130: Discovery and characterization of small molecules targeting the DNA-binding ETS domain of ERG in prostate cancer
Authors: Butler Miriam S. , Roshan-Moniri Mani , Hsing Michael , Lau Desmond , Kim Ari , Yen Paul , Mroczek Marta , Nouri Mannan , Lien Scott , Axerio-Cilies Peter , Dalal Kush , Yau Clement , Ghaidi Fariba , Guo Yubin , Yamazaki Takeshi , Lawn Sam , Gleave Martin , Gregory-Evans Cheryl Y. , McIntosh Lawrence P. , Rennie Paul S. , Cherkasov Artem , Cox Michael E.
Published: July 2017

Genomic alterations involving translocations of the ETS-related gene ERG occur in approximately half of prostate cancer cases. These alterations result in aberrant, androgen-regulated production of ERG protein variants that directly contribute to disease development and progression. This study describes the discovery and characterization of a new class of small molecule ERG antagonists identified through rational in silico methods. These antagonists are designed to sterically block DNA binding by the ETS domain of ERG and thereby disrupt transcriptional activity. We confirmed the direct binding of a lead compound, VPC-18005, with the ERG-ETS domain using biophysical approaches. We then demonstrated VPC-18005 reduced migration and invasion rates of ERG expressing prostate cancer cells, and reduced metastasis in a zebrafish xenograft model. These results demonstrate proof-of-principal that small molecule targeting of the ERG ETS domain can suppress transcriptional activity and reverse transformed characteristics of prostate cancers aberrantly expressing ERG. Clinical advancement of the developed small molecule inhibitors may provide new therapeutic agents for use as alternatives to, or in combination with, current therapies for men with ERG-expressing metastatic castration-resistant prostate cancer.

Citation Format: Miriam S. Butler, Mani Roshan-Moniri, Michael Hsing, Desmond Lau, Ari Kim, Paul Yen, Marta Mroczek, Mannan Nouri, Scott Lien, Peter Axerio-Cilies, Kush Dalal, Clement Yau, Fariba Ghaidi, Yubin Guo, Takeshi Yamazaki, Sam Lawn, Martin Gleave, Cheryl Y. Gregory-Evans, Lawrence P. McIntosh, Paul S. Rennie, Artem Cherkasov, Michael E. Cox. Discovery and characterization of small molecules targeting the DNA-binding ETS domain of ERG in prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 130. doi:10.1158/1538-7445.AM2017-130
Abstract 1309: Association between insulin resistance and breast cancer risk: A Mendelian randomization analysis of data from 228,000 women of European descent
Authors: Shu Xiang , Wu Lang , Khankari Nikhil K. , Michailidou Kyriaki , Bolla Manjeet K. , Wang Jean , Dennis Joe , Shu Xiao-ou , Simard Jacques , Easton Douglas F. , Zheng Wei
Published: July 2017

Background: Epidemiologic studies suggest that insulin resistance may be associated with breast cancer risk. We conducted Mendelian randomization (MR) analyses to reduce the biases associated with previous studies and provide evidence for causal inference.

Materials and Methods: We used genetic variants identified in genome-wide association studies for circulating fasting insulin (15 variants), early insulin secretion (16 variants), fasting proinsulin (8 variants), fasting glucose (35 variants), and 2-hour glucose (8 variants) as instruments in MR analyses. To reduce possible pleiotropic effects, variants associated with obesity were removed from the instruments. We first evaluated the association of these instruments with type 2 diabetes risk in 110,452 subjects to assess instrument validity. We then investigated the association of these instruments with breast cancer risk using data obtained from 122,977 cases and 105,974 controls of European descent included in the Breast Cancer Association Consortium (BCAC). Odds ratios (OR) were calculated to measure the associations of instrumental variables with risk of overall breast cancer and its subtypes defined by estrogen-receptor [ER] status.

Results: All instrumental variables constructed for this study were strongly associated with type 2 diabetes risk with ORs of 3.01 (p=7.86x10-5), 0.22 (p=3.54x10-14), 1.90 (p=8.28x10-4), 6.11 (p=3.59x10-19), and 1.91 (p=6.8x10-16) for per unit increase of fasting insulin, early insulin secretion, fasting proinsulin, fasting glucose, and 2-hour glucose levels, respectively. Statistically significant associations with overall breast cancer risk were found for fasting insulin (OR=1.36 for per unit increase, 95% CI=1.09-1.70, p=0.011) and fasting proinsulin (OR=1.21, 95% CI=1.06-

1.38, p=0.011). These associations were observed only for ER-positive breast cancer. No statistically significant association at p<0.05 was found for early insulin secretion, fasting glucose, or 2-hour glucose levels.

Conclusions: Our study provides strong support that certain insulin resistance traits may be causally associated with risk of breast cancer, particularly ER-positive breast cancer.

Citation Format: Xiang Shu, Lang Wu, Nikhil K. Khankari, Kyriaki Michailidou, Manjeet K. Bolla, Jean Wang, Joe Dennis, Xiao-ou Shu, Jacques Simard, Douglas F. Easton, Wei Zheng. Association between insulin resistance and breast cancer risk: A Mendelian randomization analysis of data from 228,000 women of European descent [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1309. doi:10.1158/1538-7445.AM2017-1309
Abstract 1313: Characterization of germline susceptibility to male breast cancer
Authors: Perrakis Eleni , Maguire Sarah , Tomczyk Katarzyna , Swerdlow Anthony , Orr Nick
Published: July 2017

Male breast cancer (MBC) accounts for approximately 1% of all breast cancer cases. Family history is an established risk factor for MBC and 10% of cases have germline BRCA2 mutations. We recently demonstrated that common germline polymorphisms contribute to MBC predisposition and that there is overlap between genetic susceptibility loci for MBC and female breast cancer (FBC).

Here we report a comprehensive evaluation of all 107 known FBC predisposition loci in an association analysis of 1,032 MBC cases and 2,795 population matched controls. A total of 21 FBC predisposition single nucleotide polymorphisms (SNPs) showed evidence of association with risk of MBC. For approximately a third of these SNPs, the risk estimates were significantly larger in MBC than FBC. The magnitude of effects observed suggest that common susceptibility variants may be of value for risk stratification in men who have a high absolute risk of breast cancer such as BRCA2 mutation carriers. Although our data suggests that MBC predisposition shares many features in common with FBC, there are striking differences. Surprisingly, given the predominance of hormone receptor positive tumors in MBC, we observed no evidence of association between SNPs at FGFR2 and MBC risk suggesting that the estrogen receptor status of MBC tumors does not explain the observed SNP associations.

In summary, the data presented in this abstract significantly advance our understanding of genetic factors that contribute to MBC predisposition and further underscore the growing consensus that male breast cancer is not simply analogous to hormone receptor positive female breast cancer.

Citation Format: Eleni Perrakis, Sarah Maguire, Katarzyna Tomczyk, Anthony Swerdlow, Nick Orr. Characterization of germline susceptibility to male breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1313. doi:10.1158/1538-7445.AM2017-1313
Abstract 1316: Germline genetic signals across multiple aggressive prostate cancer phenotypes
Authors: Tai Caroline G. , Emami Nima C. , Hoffmann Thomas J. , Sakoda Lori C. , Jorgenson Eric , Habel Laurel A. , Shan Jun , Ranatunga Dilrini K. , Chao Chun R. , Ghai Nirupa R. , Aaronson David , Presti Joseph , Schaefer Catherine , Risch Neil , Eeden Stephen K. Van Den , Witte John S.
Published: July 2017

Prostate cancer (PrCa) is documented to be highly heritable. But many known prostate cancer loci are not associated with aggressive prostate cancer phenotypes such as high-grade cancer or biochemical failure. Previous efforts have been largely underpowered or not replicated.

We conducted a study of 6,321 men with PrCa and 5,663 controls of European ancestry recruited from Kaiser Permanente (KP) matched by age and self-reported race. Phenotype data combined patient-level demographic, clinical, and tumor data from KP databases with data from 1994 to 2016. High-grade PrCa was defined using Gleason score: low-grade defined as ≤ 7 (3+4), high-grade defined as ≥ 7 (4+3). Biochemical failure was defined for cases using post-treatment prostate specific antigen (PSA) values: post-surgery failure if PSA ≥ 0.4 ng/mL; post-radiation failure if PSA value ≥ 2 ng/mL + nadir; and no failure if PSA did not reach specified threshold. Early-onset disease was defined as age < 55 years at diagnosis. Men were genotyped for over 650,000 SNPs using an ethnic-specific Affymetrix Axiom array and imputed to 1000 Genomes. We tested genetic associations for multiple aggressive phenotypes (410 early-onset cases, 1324 biochemical failure cases, and 886 high grade cases) in both a case-only (e.g., high vs. low grade) and case-control (e.g., high grade vs. no prostate cancer controls) analysis using logistic regression adjusting for ancestry principal components, age, BMI, and kit for batch effects. PrCa specific mortality was too rare to be evaluated.

We identified several known and novel loci. Case-control analyses highlighted known prostate cancer loci at 8q24, 10q11, 11q13, 16q22, and 19q13. Previous reports of case-only analyses also identified 8q24, 20p13 and the gene, KLK3 in 19q13, to be associated with high-grade PrCa; we confirmed these signals in our study. Previous family-based studies identified 15q11 as a familial or early-onset prostate cancer locus.

These aggressive-specific variants could perform better in predicting clinically actionable prostate cancers than variants for overall prostate cancer and can improve the predictive ability of pre-diagnostic PSA.

Citation Format: Caroline G. Tai, Nima C. Emami, Thomas J. Hoffmann, Lori C. Sakoda, Eric Jorgenson, Laurel A. Habel, Jun Shan, Dilrini K. Ranatunga, Chun R. Chao, Nirupa R. Ghai, David Aaronson, Joseph Presti, Catherine Schaefer, Neil Risch, Stephen K. Van Den Eeden, John S. Witte. Germline genetic signals across multiple aggressive prostate cancer phenotypes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1316. doi:10.1158/1538-7445.AM2017-1316