bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 324: High throughput triculture: A breast cancer spheroid model for drug screening
Authors: Benton Gabriel J. , DeGray Gerald , Arnaoutova Irina , Kleinman Hynda K. , George Jay 
Published: August 2015

Abstract
Many drugs that show promise in preclinical evaluation fail miserably during clinical trials, and the inability of current preclinical models to properly predict drug response may be attributed to oversimplified or non-existent tumor microenvironments. To provide a more physiologically predictive model for drug screening, we have developed an in vitro, 384 well, triculture system that presents key chemical (i.e. low pH, low glucose, low oxygen), extracellular matrix (ECM), and cellular (tumor cells, stromal cells and endothelial cells) attributes of tumor tissues. By using ECM proteins, we can promote the proper physiology for each of these cell types in triculture. To evaluate interactions between each cell type, they are fluorescently labeled with fluorophores with different excitation and emission spectra; MCF7 and MDA-MB-231 human breast cancer cell lines express a red fluorescence protein, while human umbilical vein endothelial cells (HUVECs) and human adipose-derived mesenchymal stem cells (hMSCs) are labeled with stable lipophilic membrane dyes. Breast tumor microtissues spontaneously assemble when breast cancer cells are co-cultured with hMSCs under low adhesion conditions. Once formed, these microtissues are deposited onto preformed tubule networks comprised of HUVECs and hMSCs and are embedded within a hydrogel composed of ECM proteins. Cellular interactions and dissemination are monitored via fluorescence microscopy, and cell proliferation of the breast cancer cells is quantified using a fluorescence plate reader. The breast cancer cells exhibit cell-cell interactions with endothelial tubules and stromal cells, forming microtissues. Furthermore, the response to anti-cancer drugs, paclitaxel and fluorouracil, in an in vitro triculture system is similar to the response for the same drugs in xenografts model, unlike 2D or 3D monoculture. This high throughput triculture cancer model will provide more insight into pathological mechanisms and can better predict response to anti-cancer drug treatment.
Citation Format: Gabriel J. Benton, Gerald DeGray, Irina Arnaoutova, Hynda K. Kleinman, Jay George. High throughput triculture: A breast cancer spheroid model for drug screening. [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 324. doi:10.1158/1538-7445.AM2015-324
Abstract 3240: Understanding clonal complexity of a tumor xenograft model via cellular barcoding technology
Authors: Caushi Justina X. , Bhang Hyo-eun C. , Li Jie , Kao Iris , Krishnamurthy Radhakrishna Viveksagar , Cooke Vesselina G. , Korn Joshua M. , Ruddy David A. , Kasibhatla Shailaja , Stegmeier Frank 
Published: August 2015

Abstract
Recent advances in next-generation sequencing have revealed the presence of genetic heterogeneity and clonal evolution within tumors. Intratumoral heterogeneity has been implicated in the disease progression, metastasis, therapeutic responses and development of drug resistance. Although cancer cell line xenograft models have been extensively used in cancer research to test drug efficacy, it is unknown how much clonal heterogeneity is maintained in the process of cell line xenograft establishment. In order to quantitatively assess the clonal complexity in xenograft models, here we applied a cellular barcoding technology using the HCC827 cell line. HCC827 is a non-small cell lung cancer cell line containing an exon 19 deletion which has been shown to be clinically relevant due to its initial response to EGFR inhibitors followed by resistance. This barcoding tool allowed us to label each individual cell with one unique DNA barcode via lentiviral infection and monitor the clonal heterogeneity within the implanted cell population by quantifying the number of unique barcodes. We were able to estimate the percentage of implanted clones that actually contributed to the formation of cell line xenografts. This study provides valuable insight on the clonal diversity present in xenograft models, which will further elucidate the heterogeneous nature of tumors.
Citation Format: Justina X. Caushi, Hyo-eun C. Bhang, Jie Li, Iris Kao, Viveksagar Krishnamurthy Radhakrishna, Vesselina G. Cooke, Joshua M. Korn, David A. Ruddy, Shailaja Kasibhatla, Frank Stegmeier. Understanding clonal complexity of a tumor xenograft model via cellular barcoding technology. [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 3240. doi:10.1158/1538-7445.AM2015-3240
Abstract 3241: Tumor-targeting Salmonella typhimurium A1-R inhibits peritoneal dissemination of ovarian cancer and prolongs survival of the tumor-bearing nude mice
Authors: Matsumoto Yasunori , Miwa Shinji , Zhao Ming , Zhang Yong , Yano Shuya , Uehara Fuminari , Yamamoto Mako , Hiroshima Yukihiko , Toneri Makoto , Bouvet Michael , Matsubara Hisahiro , Hoffmann Robert 
Published: August 2015

Abstract
Peritoneal dissemination of cancer arising from an intra-abdominal organ has poor prognosis and is treatment resistant. A peritoneal dissemination nude mouse model of ovarian cancer was made with the human ovarian cancer cell line SKOV-3-GFP. Tumor-targeting Salmonella typhimurium A1-R (A1-R) was administrated i.v. or i.p. to the tumor-bearing nude mice. Survival of the tumor-bearing nude mice was prolonged by i.v. administration of A1-R (p = 0.025) as well as by i.p. administration of A1-R (p = 0.00003), compared with the untreated control. There was no significant difference in survival between i.v. and i.p. administration (p = 0.22). However, i.p. administration was less toxic. The present results suggest A1-R has clinical potential for what is currently a highly-treatment-resistant disease.
Citation Format: Yasunori Matsumoto, Shinji Miwa, Ming Zhao, Yong Zhang, Shuya Yano, Fuminari Uehara, Mako Yamamoto, Yukihiko Hiroshima, Makoto Toneri, Michael Bouvet, Hisahiro Matsubara, Robert Hoffmann. Tumor-targeting Salmonella typhimurium A1-R inhibits peritoneal dissemination of ovarian cancer and prolongs survival of the tumor-bearing nude mice. [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 3241. doi:10.1158/1538-7445.AM2015-3241
Abstract 3243: Notch inhibitors and chemoradiation in an orthotopic glioblastoma model
Authors: Yahyanejad Sanaz , Granton Patrick , van Hoof Stefan , Barbeau Lydie , Theys Jan , Verhaegen Frank , Vooijs Marc 
Published: August 2015

Abstract
Glioblastoma multiforme (GBM) is the most common and malignant brain tumor in adults. Aggressive multimodal treatment using surgery followed by radiotherapy and chemotherapy extends the median survival of GBM patients to approximately one year after diagnosis. Treatment is not curative because of the intrinsic and acquired radiation resistance of a subpopulation of tumor cells. Notch inhibition has been shown to impair the tumorigenic capacity of these cells as well as enhance their sensitivity towards radiation. Therefore, we investigated if a highly potent and clinically approved Notch pathway inhibitor (g-secretase inhibitor) improves tumor control when combined with radiotherapy and chemotherapy in an orthotopic GBM mouse model.
To investigate the treatment efficacy of combinations treatments between standard of care treatment (radiotherapy and temozolomide) and Notch inhibitors, we used a three dimensional spheroid growth assay using both established and primary human glioma cell lines in which spheroid volume growth delay was quantitatively monitored. In addition, we assessed the expression of the putative glioma stem cell marker CD133 using flow cytometry. Furthermore, therapeutic efficacy of these combination treatments was als assessed in vivo in an orthotopic glioma tumor model (U87-luc) wherein tumor progression was evaluated using bioluminescence (BLI) and contrast-enhanced micro-computed tomography (CT) imaging. A small animal precision irradiation platform (PXI, XRAD 225Cx, CT, USA) was used for micro-CT imaging and irradiation delivery of conformal doses between 2- 10 Gy to intracranial tumors in mice receiving single or combination treatments with Notch inhibitors and chemotherapy.
Results: Notch blockade alone did not affect the sphere volume compared to control, whereas combination treatment with radiation and / or temozolomide resulted in a substantial spheroid growth delay (p = 0.004). Irradiation enhanced the expression of the stem cell marker CD133, while Notch blockade reduced CD133 expression. In intracranial tumours we found a strong correlation between CT and BLI imaging of tumor growth (Pearson coefficient (r) = 0.85, p = 0.001). The potential of Notch inhibition combined with precision radiotherapy and chemotherapy in our orthotopic GBM model is currently being determined and the outcome of this study will be presented.
Citation Format: Sanaz Yahyanejad, Patrick Granton, Stefan van Hoof, Lydie Barbeau, Jan Theys, Frank Verhaegen, Marc Vooijs. Notch inhibitors and chemoradiation in an orthotopic glioblastoma 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 3243. doi:10.1158/1538-7445.AM2015-3243
Abstract 3246: Novel luciferase labelling technique to improve imaging of orthotopic prostate and pancreatic cancer models
Authors: Bernoulli Jenni , Tuomela Johanna , Bozza Matthias , Fagerlund Katja M. , Suominen Mari I. , Morris George , Halleen Jussi M. , Harbottle Richard 
Published: August 2015

Abstract
Bioluminescent-labelling allows sensitive non-invasive sequential imaging of tumor development and early metastasis. However, current methods for the genetic modification of cells typically use integrating genotoxic viruses that can potentially disrupt the molecular behavior of cancer cell lines due to their random nature of integration. Here, we utilized a non-viral DNA vector that comprises an S/MAR (Scaffold/Matrix Attachment Region) element to stably modify cells to be further used in xenograft studies to allow long term expression without affecting cell behavior or silencing over cell divisions. Human PC-3 prostate cancer cells and BxPC-3 pancreatic cancer cells were stably transfected with a pCAG-LUC-S/MAR and cultured for 4 weeks under selection. Colonies that formed after this period were isolated and expanded in normal medium and evaluated for luciferase expression and molecular integrity of the DNA vector. For in vivo studies, PC-3 cells were inoculated orthotopically into the prostate, and BxPC-3 cells into the pancreas using athymic and BALB/c nude mice. For comparison, similar experiments with the corresponding study designs were performed with the non-labelled parental cell lines. The luciferase transfected cells maintained their original properties with stable expression. Luciferase-labelled PC-3 and BxPC-3 cells inoculated orthotopically into the prostate and pancreas, respectively, were successfully followed for 5 weeks with non-invasive bioluminescence imaging by IVIS. The results demonstrated high-quality follow-up of tumor growth compared with tumor models using non-labelled cells. In conclusion, S/MAR DNA vectors are able to produce genetically modified cells without the limitations of random genomic integration, whilst providing extra-chromosomal mitotic stability and sustained transgene expression at high level. When utilized in orthotopic xenograft studies, these luciferase expressing cells formed a reliable and essential non-invasive imaging platform that improves substantially efficacy testing of anticancer drug candidates.
Citation Format: Jenni Bernoulli, Johanna Tuomela, Matthias Bozza, Katja M. Fagerlund, Mari I. Suominen, George Morris, Jussi M. Halleen, Richard Harbottle. Novel luciferase labelling technique to improve imaging of orthotopic prostate and pancreatic cancer models. [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 3246. doi:10.1158/1538-7445.AM2015-3246
Abstract 3251: Regulation of TNBC kinome by the metastasis suppressor Raf kinase inhibitory protein (RKIP)
Authors: Yesilkanal Ali E. , Frankenberger Casey , Rabe Daniel , Johnson Gary L. , Rosner Marsha R. 
Published: August 2015

Abstract
Triple negative breast cancer (TNBC) is the most aggressive subtype of breast cancer with very low survival rates. Metastatic progression of tumors is the major cause of death in TNBC patients. Metastatic spread of tumor cells is generally associated with resistance to therapy and poor prognosis. Therefore, there is a clinical need for understanding the molecular and cellular basis of metastasis and developing effective therapies against metastatic progression. However, since metastasis is a multi-step process, unraveling its complexity is a major challenge. One effective way of tackling this question is to study natural blockers of the metastatic process, metastasis suppressors, and identify the mechanisms by which they regulate metastasis. Raf kinase inhibitory protein (RKIP) is a metastasis suppressor in TNBC, which acts as a homeostatic factor that regulates kinase activity. RKIP blocks invasion in vitro and experimental metastasis in vivo. Therefore, RKIP constitutes a powerful tool for studying metastatic signaling pathways in cancer cells. RKIP is known to regulate the activity of certain kinases such as Raf-1, GRK2, NIK, and GSK3β, affecting major pathways such as MAPK, NFκB, and WNT. However, our preliminary data suggests that RKIP affects a much larger part of the TNBC kinome. Using proteomics approaches, we have identified putative novel kinase targets of RKIP, and used bioinformatics analysis to build RKIP-regulated signaling networks. Elucidating RKIP's function at a systems level will reveal the interplay between key metastatic signaling cascades and potentially identify novel anti-metastatic target combinations in TNBCs.
Citation Format: Ali E. Yesilkanal, Casey Frankenberger, Daniel Rabe, Gary L. Johnson, Marsha R. Rosner. Regulation of TNBC kinome by the metastasis suppressor Raf kinase inhibitory protein (RKIP). [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 3251. doi:10.1158/1538-7445.AM2015-3251
Abstract 3261: Cytochrome p450 3a5 manipulates the progression of hepatocellular carcinoma via selectively targeting ros/mtorc2/p-akt (s473) signaling
Authors: chen lei , Wang hongyang 
Published: August 2015

Abstract
CYP3A5, one of the cytochrome P450 superfamily members, is involved in the metabolism of drugs, carcinogens, and endogenous molecules. Previous studies mainly focused on the role of CYP3A5 polymorphism or altered enzyme activity during carcinogenesis. However, the potential physiological function of CYP3A5 in tumor progression remains largely unknown. In our present study, the expression of CYP3A5 was firstly reported frequently down-regulated in not only 159 clinical HCC samples but also 4 external HCC cohort(n = 695). Tumors with reduced level of CYP3A5 were more likely to have aggressive characteristics (vascular invasion and poor differentiation), together with the worse overall survival time and time to disease recurrence. The exogenous expression of CYP3A5 was associated with the induction of TIMP1/2 and dysfunction of MMP2/9, which suppressed HCC migration and invasion in vitro and in vivo mechanically through inhibiting AKT signaling. Strikingly, the selective inhibition of AKT phosphorylation at Ser473(instead of Thr308) residue was found in CYP3A5-stable cells with an mTORC2-dependent manner. Co-immunoprecipitation and kinase assay further confirmed that mTORC2 kinase activity rather than Rictor/mTOR complex integrity was necessary for the inhibition of p-AKT(S473). Importantly, CYP3A5-induced ROS accumulation was identified as the key factor for the regulation of mTORC2 activity, in line with the decreased GSH/GSSG contents in most clinical HCC samples with relatively lower metastatic capacity. Together, these findings highlighted the novel function of CYP3A5 in the regulation of tumor metastasis via enhancing ROS level and in turn arresting mTORC2/p-AKT(S473) signaling, which may serve as a potential prognostic biomarker and therapeutic target for HCC.
Citation Format: lei chen, hongyang Wang. Cytochrome p450 3a5 manipulates the progression of hepatocellular carcinoma via selectively targeting ros/mtorc2/p-akt (s473) signaling. [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 3261. doi:10.1158/1538-7445.AM2015-3261
Abstract 3275: A novel approach to treat medulloblastoma: The omega-3 fatty acids DHA and EPA reduce medulloblastoma tumor growth in vitro and in vivo
Authors: Ljungblad Linda M. , Johnsen John-Inge , Wickström Malin , Kogner Per , Gleissman Helena 
Published: August 2015

Abstract
Background: The omega-3 fatty acids docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) exert antitumoral properties shown in several cancer types. DHA is found in abundance in the brain and is of importance in normal brain development and maintenance of function including cognitive functions.
Medulloblastoma is the most common malignant CNS tumor of childhood. Intensive multimodal treatment is necessary including surgery, radiotherapy and chemotherapy. However, treatment often results in significant neurological sequelae and the risk of resistant relapses is still significant. The neuroprotective and antitumoral properties of omega-3 could therefore be of great benefit. Our aim was to investigate the effect of DHA and EPA on medulloblastoma in vitro and in vivo.
Methods: In a panel of medulloblastoma cell lines the cytotoxic activity of DHA and EPA was studied using cell viability assays. The molecular mechanisms were characterized using cell- and molecular biology techniques. Mice with human medulloblastoma xenografts were treated with; DHA alone or with DHA and EPA for 30 days from time of tumor take while tumor growth was monitored. Control mice were fed with normal chow. The incorporation of fatty acids was analyzed in tumors, erythrocytes and brain tissue by LS-MS/MS.
Results: DHA and EPA induced medulloblastoma cell toxicity with IC50 values ranging from 1.9 to 68 μM in six medulloblastoma cell lines. DHA reduced the prostaglandin E2 production, indicating a possible mechanism of action. In vivo, omega-3 supplementation resulted in significant reduction of tumor growth when established tumors were treated. DHA levels were increased in both tumor tissue and erythrocyte membrane compared to control. The in vivo treatment was non-toxic.
Conclusions: DHA and EPA exert toxic effect on medulloblastoma cells in vitro and reduce tumor growth in vivo. These omega-3 fatty acids could therefore be good candidates for improving current therapy and at the same time protect healthy neurons from therapy-induced toxicity promoting cognitive function in survivors.
Thus, omega-3 fatty acid supplementation has the potential of reducing tumor growth as well as reducing sequelae.
Citation Format: Linda M. Ljungblad, John-Inge Johnsen, Malin Wickström, Per Kogner, Helena Gleissman. A novel approach to treat medulloblastoma: The omega-3 fatty acids DHA and EPA reduce medulloblastoma tumor growth in vitro and in vivo. [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 3275. doi:10.1158/1538-7445.AM2015-3275
Abstract 3282: Induction and characterization of cyclopamine resistance in SK-N-DZ neuroblastoma and DAOY medulloblastoma cell lines
Authors: de la Rosa Javier , Paredes Amaia , Asensio-Salazar Javier , Fan Xing , Meléndez Bárbara , Rey Juan A. , Castresana Javier S. 
Published: August 2015

Abstract
Neuroblastoma and medulloblastoma are malignant solid tumors of childhood and early adolescence, that often reappear after clinical treatment due of an acquired chemo-resistance. These tumors present an increased expression of the Sonic Hedgehog signaling pathway. In the last years, inhibitors of this pathway, such as cyclopamine are being studied as a possible alternative to current treatment. In this research, we aim at analyzing the appearance and characteristics of resistance derived from the treatment with mutation-selective concentrations of cyclopamine. The aim of this study is the design of different ways to bypass the resistance generated by the tumor once its biology is completely understood.
Cells were exposed to several treatment cycles. A 24 h cyclopamine exposure was performed when cell culture reached 60-70% confluence. Basal half-maximal inhibitory concentration (IC50) was 10 μM. Hence, SK-N-DZ cells received 5 treatment cycles of cyclopamine at 12 μM dissolved in DMSO; whereas Daoy cells received 5 treatment cycles of cyclopamine at 10 μM. The control group only received the diluent.
Anchorage-independent growth assays (in soft agar), clonogenicity assays, cell migration scratch (wound-healing) assays, and relative mRNA expression were studied during cycles 1-5.
During the first 5 treatment cycles, mRNA expression of SMO remained constant. CD15 expression decreased. MYCN, Gli and expression showed an increasing trend. Decrease in the pro-apoptotic status was also described. Soft agar showed an increase tendency in relative colony formation capacity. No statistical differences were found in the clonogenicity assays for the SK-ND-Z cells.
The results highlighted the evidence that cells became resistant because they were able to grow in soft agar without attachment, to migrate and to overexpress genes related to cancer stem cells. Results shown are congruent with a resistance generation possibly owing to multidrug efflux pumps. However, more clear results would be seen with 48 h treatment. Further analysis based on this work could include 48 h treatment, water-dissolvable cyclopamine performance study, a more complete gene analysis, or studies with other neurological tumors.
Note: This abstract was not presented at the meeting.
Citation Format: Javier de la Rosa, Amaia Paredes, Javier Asensio-Salazar, Xing Fan, Bárbara Meléndez, Juan A. Rey, Javier S. Castresana. Induction and characterization of cyclopamine resistance in SK-N-DZ neuroblastoma and DAOY medulloblastoma cell lines. [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 3282. doi:10.1158/1538-7445.AM2015-3282