bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3080: Definition of the Ewing sarcoma specific oncomir-1 targetome
Authors: Schwentner Raphaela , Herrero-Martin David , Kauer Maximilian , Kovar Heinrich 
Published: August 2015

Abstract
MicroRNAs (miRNA) serve to fine-tune gene expression and thus play an important regulatory role in tissue specific gene networks. The identification and validation of miRNA target genes in a given tissue still poses a significant problem, since the presence of a seed sequence in the 3´UTR of an mRNA and its expression modulation upon forced ectopic expression of the miRNA do not reliably predict regulation under physiological conditions. Here, we used a combination of AGO2 pull-down experiments by PAR-CLIP - to enrich and sequence RISC-associated RNAs - and of RNAseq upon miRNA depletion by ectopic sponge expression, to identify the targetome of miR17-92 in Ewing sarcoma (ES) cells. The chimeric oncoprotein EWS-FLI1 is the driving pathogenic force in ES. As an aberrant ETS transcription factor it directly deregulates a multitude of genes related to proliferation control and differentiation. Recently, we reported on the EWS-FLI1 microRNA (miRNA) signature based on knockdown experiments in ES cell lines and on differential expression in primary tumors versus mesenchymal stem cells. Comparison of both datasets revealed miR20a-3p to be the top EWS-FLI1 activated miRNA. MiR-20a belongs to the miR17-92 cluster, also known as oncomir-1, which is one of the most potent oncogenic miRNAs. The whole cluster is among the small number of miRNAs down-regulated upon knockdown of EWS-FLI1 in multiple ES cell lines. Since feedback loops exists between the E2F transcription factor family and miR-17-92 and between EWS-FLI1 and E2F3, we sought to investigate the role of this miRNA cluster in ES.
We found a significant enrichment of PAR-CLIP hits for members of the miR-17-92 cluster, in the 3´UTRs of genes up-regulated in response to mir-17-92 specific sponge expression. Among them we consistently identified known (i.e. CTGF, MXD1, CYLD1) and a multitude of so far unknown targets of the oncomir-1 cluster in ES in three PAR-CLIP experiments. Preliminary experiments using 3′UTRs of select novel candidate genes in gene reporter assays showed up-regulation of luciferase activity upon transfection of a sponge specific to miR-17-92 and 20a, but not to unrelated miR-9 and 10b. Site directed mutagenesis of seed sequences in the 3′UTR of these candidate genes will be used for further validation.
This study provides an example for the comprehensive definition of the targetome for a defined group of miRNAs under physiological conditions.
This study was supported by the 7th framework program of the European Commission (FP7-259348, “ASSET”) and the Austrian Science Fund FWF (24708-B21)
Citation Format: Raphaela Schwentner, David Herrero-Martin, Maximilian Kauer, Heinrich Kovar. Definition of the Ewing sarcoma specific oncomir-1 targetome. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3080. doi:10.1158/1538-7445.AM2015-3080
Abstract 3096: CDK4/6 as a therapeutic target in malignant melanoma
Authors: Teh Jessica L. , Aplin Andrew 
Published: August 2015

Abstract
Melanoma is the most deadly form of skin cancer and FDA-approved therapies such as vemurafenib and ipilimumab are associated with short-term responses and/or severe toxicities for patients. Recent favorable clinical data with cyclin-dependent kinase (CDK) 4/6 inhibitor, palbociclib (PD’991) in ER+ breast cancer has rekindled an interest in targeting cell cycle progression to stop aberrant cell growth. Thus, we set out to analyze the potential use of CDK4/6 inhibitors in melanoma. Due to the inability of CDK4/6 inhibitors to induce apoptosis as a single agent, we explored viable combinatorial strategies that will trigger both pro-senescent and pro-apoptotic pathways that may lead to an effective treatment modality for melanoma patients. Here, we demonstrate the benefit of combining a MEK inhibitor, trametinib (GSK’212) with PD’991 both in vitro and in vivo. We did not observe any enhanced effects of combining both GSK’212 and PD’991 in cells that are wild-type for BRAF and NRAS or an RB1-null BRAF mutant line. However, B-RAF and N-RAS mutant cells exhibited an enhanced response to the combinatorial agents as compared to single agents. Furthermore, we observed a dramatic increase in apoptosis in cell lines that are sensitive to combined GSK’212 and PD’991 treatment. Expression of Inhibitor-of-Apoptosis (IAP) protein, survivin, was associated with response. Finally, because it is important to understand how resistance develops, we have validated a novel E2F reporter system in vivo to quantitatively and temporally measure the efficacy of CDK4/6 inhibitors and possible re-activation of the pathway during acquired drug resistance.
Citation Format: Jessica L. Teh, Andrew Aplin. CDK4/6 as a therapeutic target in malignant melanoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3096. doi:10.1158/1538-7445.AM2015-3096
Abstract 3103: Cyclin E amplification predicts sensitivity of primary Uterine Serous Carcinoma (USC) cell lines to the cdk2 inhibitor CYC065
Authors: Cocco Emiliano , Bellone Stefania , Lopez Salvatore , Bonazzoli Elena , Predolini Federica , Black Jonathan D. , Santin Alessandro D. 
Published: August 2015

Abstract
We evaluated the in vitro effectiveness of the cdk2 inhibitor CYC065 on multiple primary chemotherapy-resistant USC cell lines with or without CCNE1 amplification.
CCNE1-amplified primary cell lines were significantly more sensitive than wild type USC cell lines to CYC065 in vitro (i.e., IC50: mean±STDV = 61.75±13.22 nM and 103.16± 21.9 nM for CCNE1-amplified USC-ARK-2 and USC-ARK-7 cell lines, respectively and 539.2±182.1 nM for the wild type USC-ARK-4 cell line, p = 0.0048). Consistently, low concentrations of CYC065 (i.e., 100 - 300 nM) caused a dose dependent arrest in the G1 phase of the cell cycle specifically in CCNE1-amplified primary USC cell lines. Importantly, CCNE1 knockdown in the USC-ARK-2 cell line resulted in a 9.29-fold increase in the CYC065 IC50 when compared to the MOCK-transfected USC-ARK-2 cell line (p = 0.021). Finally, when primary CCNE1-amplified USC cell lines also harboring ERBB2 amplification (50% of CCNE1-amplified USC cell lines) were incubated in vitro with the combination of CYC065 and Herceptin (a monoclonal antibody targeting the product of the ERBB2 gene, HER2/neu), an increased inhibitory effect was reported in the combination treatment when compared to Herceptin or CYC065 used as single agent (i.e.,% viable cells: mean±STDV = 71.4±0.85, 65.4±14.2, 42.2±2.1 for the Herceptin, the CYC065 and the combination treatment on USC-ARK-2, respectively; p = 0.014).
Together these findings identify CYC065 as a promising drug to be considered alone or in combination in the treatment of patients harboring CCNE1-amplified USC.
Citation Format: Emiliano Cocco, Stefania Bellone, Salvatore Lopez, Elena Bonazzoli, Federica Predolini, Jonathan D. Black, Alessandro D. Santin. Cyclin E amplification predicts sensitivity of primary Uterine Serous Carcinoma (USC) cell lines to the cdk2 inhibitor CYC065. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3103. doi:10.1158/1538-7445.AM2015-3103
Abstract 3106: Elucidating molecular mechanisms linking microRNA-206 loss to tumor progression and metastasis
Authors: Watt Kathleen D. , Truesdell Peter , Craig Andrew W. 
Published: August 2015

Abstract
Metastasis is the leading cause of cancer-related deaths, and this occurs when tumor cells invade basement membranes and blood vessels to colonize other tissues. Metastasis is facilitated by extracellular matrix (ECM)-degrading structures called invadopodia. Recently, several microRNAs (miRNAs) have been identified as tumor and metastasis suppressor genes. Loss of expression of miR-206 has been correlated with poor prognosis in gastric cancer, breast cancer, rhabdomyosarcoma and melanoma. A previous study by others has demonstrated that restoring miR-206 expression in human breast cancer cells was sufficient to block metastasis in tumor xenograft assays. However, the targets of miR-206 that cause metastasis in these cancer models have not been fully characterized. Here, we have tested the effects of miR-206 on invadopodia in metastatic cancer cells, and identified a novel target that may explain this effect. Transducer of Cdc42-mediated Actin assembly-1 (Toca-1) is an adaptor protein that promotes formation of invadopodia and metastasis in breast cancer models, and has a predicted miR-206 binding site within the 3′ UTR of Toca-1 transcripts. To test whether the high levels of Toca-1 expression that we have observed in metastatic cancer cell lines was due to loss of miR-206, we rescued miR-206 expression using transient, stable or inducible approaches in MDA-MB-231 breast cancer, A375 melanoma, and H1299 lung cancer cell lines. In each cell model, Toca-1 expression was reduced upon miR-206 expression at both the mRNA and protein levels, thus validating Toca-1 as a new target of miR-206. Further testing of the phenotypes induced by miR-206 rescue revealed dramatic defects in ECM degradation and cell invasion in breast cancer and melanoma cell models. In subcutaneous tumor xenograft assays, stable rescue of miR-206 in H1299 tumors led to defects in tumor growth compared to those expressing a scrambled control miRNA. In addition, miR-206 expression caused a dramatic reduction in the numbers of lung metastases. We are currently extending this to our melanoma models and inducible models to define changes in metastasis-related targets of miR-206 that are aberrantly expressed in metastatic cancers. In conclusion, our study demonstrates for the first time a link between miR-206 and suppression of invadopodia formation via Toca-1 silencing, which likely contributes to the metastasis suppressing activity of miR-206 in multiple cancer types.
Citation Format: Kathleen D. Watt, Peter Truesdell, Andrew W. Craig. Elucidating molecular mechanisms linking microRNA-206 loss to tumor progression and metastasis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3106. doi:10.1158/1538-7445.AM2015-3106
Abstract 311: High-throughput spheroid formation in a 384-well format using magnetic 3D bioprinting
Authors: Tseng Hubert , Gage Jacob A. , Haisler William L. , Souza Glauco R. 
Published: August 2015

Abstract
Technical limitations in the rapid and reproducible formation of multicellular tumor spheroids prevent their widespread use in high-throughput screening for cancer research and drug development. Existing systems for spheroid formation require lengthy processing times, and make simple tasks like media exchange and sample retention challenging. An ideal 3D cell culture system would print spheroids rapidly, while being easy to handle and process for high-throughput applications.
To address this unmet need, we use magnetic 3D bioprinting for high-throughput spheroid formation. Magnetic 3D bioprinting is based on the principle of magnetizing cells using a magnetic nanoparticle assembly. Once magnetized, these cells can be directed and aggregated using magnetic forces; specifically, magnetized cells can be aggregated into spheroids in 384-well plates. These spheroids can be cultured long-term, and, as they are magnetic, the spheroids can be held down or transferred out of the well, avoiding the technical issues of other 3D systems to retain samples. Moreover, the magnetic nanoparticles have no effect on cell behavior and do not interfere with fluorescence.
We demonstrated this method by printing spheroids of HepG2, HCT-116, A549, and PANC-1 cancer cells in 384-well plates. We printed spheroids rapidly (15 min) and reproducibly, with cell numbers as small as 100 cells. We assayed the spheroids by using their contraction over time as an endpoint, which we validated as a toxicity endpoint correlating with cell migration, proliferation, and viability. In running these experiments we also demonstrated that these spheroids are amenable to high-content testing, with multiple experiments being performed on the same spheroid to yield more data per test.
Thus, this study introduces magnetic 3D bioprinting for high-throughput spheroid formation, where multicellular tumor spheroids that represent native tumor environments can be rapidly and easily printed for cancer research and compound screening.
Citation Format: Hubert Tseng, Jacob A. Gage, William L. Haisler, Glauco R. Souza. High-throughput spheroid formation in a 384-well format using magnetic 3D bioprinting. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 311. doi:10.1158/1538-7445.AM2015-311
Abstract 3117: Hsa-miR-140-5p inhibits colorectal cancer stem cell invasion and metastasis by suppressing Smad2 and autophagy
Authors: Zhai Haiyan , Fesler Andrew , Ju Jingfang 
Published: August 2015

Abstract
Colorectal cancer is one of the most common cancers and frequently metastasizes to the liver. TGF-β signaling has been shown to regulate cancer metastasis by promoting epithelial to mesenchymal transition and cancer stem cell (CSC) phenotypes. Smad2 is a key element downstream of TGF-β. In this study, we showed that hsa-hsa-miR-140-5p-5p directly targets Smad2 and overexpression of hsa-miR-140-5p in colorectal cancer cell lines decreases Smad2 expression level, which leads to decreased level of cell invasion and proliferation, as well as increasing cell cycle arrest. Ectopic expression of hsa-miR-140-5p in colorectal CSCs inhibited CSC growth and sphere formation in vitro by disrupting autophagy, an essential process for cancer cell survival and chemoresistance. We have systematically identified the targets of hsa-miR-140-5p involved in autophagy. Furthermore, overexpression of hsa-miR-140-5p in CSCs abolished the tumor formation as well as metastasis in vivo by using subcutaneous xenografts and tail vein injection of CSCs in NOD/SCID mice. In addition, there is a progressive loss of hsa-miR-140-5p expression from normal colorectal mucosa to primary tumor tissues, with further reduction in liver metastatic tissues, quantified by qRT-PCR. The functional and clinical significance of hsa-miR-140-5p suggests that it is a key regulator in colorectal cancer progression and metastasis. As a result, it may have potential as a novel therapeutic molecule to treat colorectal cancer.
Citation Format: Haiyan Zhai, Andrew Fesler, Jingfang Ju. Hsa-miR-140-5p inhibits colorectal cancer stem cell invasion and metastasis by suppressing Smad2 and autophagy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3117. doi:10.1158/1538-7445.AM2015-3117
Abstract 313: Examining the role of ABCA1 cholesterol transporter in ovarian cancer spheroids
Authors: Williams Rebekka , Russell Amanda , Bongers Angelika , Sagnella Sharon , Fife Christopher , Jessup Wendy , DeFazio Anna , Chenevix-Trench Georgia , Haber Michelle , Norris Murray , Henderson Michelle 
Published: August 2015

Abstract
Epithelial ovarian cancer (EOC) is a devastating disease which accounts for a large proportion of gynaecological cancer-related deaths. Poor survival is largely due to late diagnosis of the disease which typically presents with peritoneal dissemination. In order to achieve reduced tumor burden and improved survival, it is imperative to identify new prognostic markers and potential therapeutic targets. Recent work by our group identified high ATP- Binding Cassette A1 (ABCA1) transporter expression as being associated with poor outcome in EOC. ABCA1 has been widely studied as a cholesterol transporter, however there is little research investigating its potential roles in cancer biology. The aim of this work was to ascertain whether ABCA1 may promote aggressive characteristics in EOC. siRNA-mediated suppression of ABCA1 in EOC cell lines led to reduced colony forming ability, growth and migration by comparison with controls. By culturing EOC cell lines (Skov3, 27/87 and HEY) in a 3-dimensional spheroid model, we have shown that ABCA1 suppression significantly reduced overall spheroid volume (p = ≤0.0003, ANOVA with Dunnett's multiple comparison test). In Skov3 and 27/87 spheroids, reduced volume was associated with cell death; however in HEY spheroids, ABCA1 suppression significantly reduced cell numbers in the absence of detectable cell death (p = ≤0.0002, ANOVA with Dunnett's multiple comparison test). Detailed characterisation of spheroid architecture revealed an increased volume-to-cell ratio in those depleted of ABCA1, indicating a lower degree of compaction compared to controls. It was further observed that whilst HEY spheroids derived from control siRNA-transfected cells develop a prominent necrotic core this feature was dramatically impaired by ABCA1 suppression. Compaction and core necrosis are both well-documented processes in spheroid biology that are indicative of cellular contractility and a hypoxic tumour environment, both key characteristics of more aggressive, chemo-resistant and recurrent tumors. Collectively these data suggest that ABCA1 may be involved in regulating multiple processes associated with aggressive and invasive characteristics of EOC, highlighting ABCA1 as a potential therapeutic target.
Citation Format: Rebekka Williams, Amanda Russell, Angelika Bongers, Sharon Sagnella, Christopher Fife, Wendy Jessup, Anna DeFazio, Georgia Chenevix-Trench, Michelle Haber, Murray Norris, Michelle Henderson. Examining the role of ABCA1 cholesterol transporter in ovarian cancer spheroids. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 313. doi:10.1158/1538-7445.AM2015-313
Abstract 3130: Discovery of tumor suppressor microRNA using smoking-related lung carcinogenesis cell model
Authors: Sung Jae Sook , Lee Jong Won , Kim Yeul Hong 
Published: August 2015

Abstract
Cigarette smoking is an important risk factor for lung cancer and induces DNA hypermethylation. Hypermethylation can silence tumor suppressor miRNAs. However, it is less known regarding the role of miRNA methylation in lung carcinogenesis, especially smoking-related lung cancer. We are now to discover the smoking-induced altered tumor suppressor miRNA in human lung carcinogenesis cell model. First, we examined miRNA expression profiles using Affymetrix GeneChip miRNA4.0 in human lung carcinogenesis cell model; NHBE, BEAS2B, 1799, 1198, 1170I cells (PNAS. 89:6693-97. 1992, Cancer Biol Ther. 5:665-73. 2006). And we have validated candidate tumor suppressor miRNAs by TaqMan real time PCR. We first discovered 6 candidate tumor suppressor miRNAs (miR-4448, miR-3180-3p, miR-584-5p, miR-3658, miR-8064, miR-371b) and we confirmed known decreased expression of several miRNAs in lung carcinogenesis cell model. Next, we tested methylation of candidate tumor suppressor miRNAs promoter CpG site by pyrosequencing. Increase of methylation was observed for 4 of the 6 candidate tumor suppressor miRNAs in human lung carcinogenesis cell model. Furthermore, methylation of miR-584-5p, miR-3658, miR-8064, miR-371b were greater in 1198 and 1170I cells exposed to cigarette smoking condensate (CSC). In the further study, we will confirm the role of tumor suppressor miRNA in smoking-related lung carcinogenesis by functional study. These finding suggest that tumor suppressor miRNAs are miRNA silenced by epigenetic mechanisms during smoking-induced lung carcinogenesis. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2013R1A1A2062146) and the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (HI13C1611).
Citation Format: Jae Sook Sung, Jong Won Lee, Yeul Hong Kim. Discovery of tumor suppressor microRNA using smoking-related lung carcinogenesis cell model. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3130. doi:10.1158/1538-7445.AM2015-3130
Abstract 314: A rapid 3D tumor spheroid analysis method using the Celigo imaging cytometry
Authors: Chan Leo Li-Ying , Cribbes Scott , Vinci Maria , Kessel Sarah , Patterson Lisa , Eccles Sue 
Published: August 2015

Abstract
The current 2D methods for cancer drug discovery have had some difficulty in identifying potential drug candidates that can be used for clinical testing. To overcome this challenge, there has been an increase in research of 3D tissue culture that facilitated the development of new in-vitro tumor model assays. Traditional 2D and 3D analysis method relied heavily on visual observation using microscopy. However, the method is time-consuming and has high variations. Automated plate-based imaging cytometer can be employed to rapidly analyze and characterize 3D tumor spheroids, which can be used to generate both quantitative and qualitative results. In this work, we demonstrate a novel 3D tumor spheroid analysis method using the Celigo imaging cytometer for spheroid counting, size analysis, tumor migration and invasion, tumor viability, and dose response of drug induced/inhibited tumor growth. The plate-based imaging cytometer utilizes bright-field and three fluorescence channels (Blue, Green, and Red) for multi-channel analysis. By utilizing the F theta lens technology, uniform bright-field image is captured for more accurate counting of the entire well. In addition, Celigo analysis software is used to report numerous parameters allowing detailed spheroid characterization. In addition to direct spheroid counting in the well, the use of specific fluorescent dyes and probes allow the user to define viable and hypoxic areas within spheroids and monitor migration and invasion on or into supporting cells and/or tissues. The results showed that Celigo imaging cytometer can accurately count and measure spheroid sizes in response to drug induction. Furthermore, tumor migration and invasion were clearly observed and quantified in the captured images. By utilizing the 3D spheroid imaging cytometry method, researchers can quickly characterize and quantify tumor spheroids, which can improve the efficiency of identification of potential cancer drug candidates.
Citation Format: Leo Li-Ying Chan, Scott Cribbes, Maria Vinci, Sarah Kessel, Lisa Patterson, Sue Eccles. A rapid 3D tumor spheroid analysis method using the Celigo imaging cytometry. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 314. doi:10.1158/1538-7445.AM2015-314