bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3318: Phosphoproteomic profiling of glioblastoma stem cells to uncover mechanisms of radiation resistance
Authors: Bassett Emily , Palanichamy Kamalakannan , Chakravarti Arnab
Published: August 2015

Glioblastoma (GBM) is the most common and aggressive primary malignant brain tumor in adults. GBM tumors universally develop resistance to chemotherapy and radiation therapy; thus, there is an urgent need to develop novel treatment strategies. In this study, we identify signaling pathways altered specifically in radioresistant glioblastoma stem cells (GSCs) following radiation treatment, in order to identify mechanisms of radiation resistance. Protein phosphorylation is a highly regulated and dynamic post-translational modification that is the primary mechanism of signal transduction. We hypothesized that phosphoproteomic profiling of GSCs can be used to identify novel signaling pathways involved in GBM radioresistance. Phosphopeptides were isolated from three GSC lines and one differentiated GBM cell line before and after radiation treatment. Mass spectrometry-based global phosphoproteomic profiling was then utilized to quantify radiation-induced changes in protein phosphorylation. About 6,000 phosphopeptides mapping to about 2,300 phosphoproteins were quantified at two post-radiation timepoints (relative to untreated) across all four cell lines. Three biological replicates of each sample were analyzed by mass spectrometry to obtain statistical significance. Fold changes of phosphoproteins following radiation treatment were then mapped to canonical signaling pathways, and pathways altered specifically in radiation-resistant GSCs were identified. We conclude that different pathways are activated in GSCs compared to differentiated GBM cells following radiation treatment, and some of these pathways may mediate radioresistance. Future efforts will focus on validation of the mass spectrometry results in vivo, and cell-based assays to identify the role of specific phosphoproteins in radioresistance pathways. Results of this study will guide future efforts to develop radiosensitizing agents that, when used in combination with current treatments, may increase the survival of patients with this devastating disease. This study was supported by funds from The Ohio State University Comprehensive Cancer Center (to AC); NIH/NCI 1U10CA180850-01(to AC); NIH/NCI 1RC2CA148190 (to AC); NIH/NCI 2K12CA133250-06 (to AC); NIH/NCI/CTEP 1R01CA169368 (to AC); NIH/NCI 1R01CA11522358 (to AC), RTOG (to AC).

Citation Format: Emily Bassett, Kamalakannan Palanichamy, Arnab Chakravarti. Phosphoproteomic profiling of glioblastoma stem cells to uncover mechanisms of radiation resistance. [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 3318. doi:10.1158/1538-7445.AM2015-3318
Abstract 3319: Identification of cervical cancer stem cells by using an imaging system for proteasome activity
Authors: Tamari Keisuke , Ishii Hideshi , Hayashi Kazuhiko , Konno Masamitsu , Kawamoto Koichi , Nishida Naohiro , Kozeki Jun , Fukusumi Takahito , Doki Yuichiro , Mori Masaki , Ogawa Kazuhiko
Published: August 2015

Cervical cancer is one of the most common cancers among women worldwide and the fourth most common cause of cancer death in women worldwide (275,100 deaths in 2008). For advanced cervical cancer, concurrent radiotherapy and chemotherapy are commonly administered. However, the 5-year survival rate remains at only ∼60%, indicating the need for significant improvements in clinical outcomes. Recurrence and metastatic potential can be due to a specific radioresistant subpopulation of cells with stem-like characteristics, cancer stem cells (CSCs) which maintain the capacity to regenerate entire tumors. Targeting of the CSCs is a promising avenue to explore to develop new therapies for this devastating cancer. Recently, cancer cells with low proteasome activity have been identified as CSCs in breast cancer, glioma, and pancreatic cancer with a fluorescence marker system for level of proteasome activity. We stably infected two cervical cancer cell lines, HeLa and CaSki, with an expression vector for a fusion protein between the green fluorescent protein, ZsGreen, and the C-terminal degron of murine ornithine decarboxylase to monitor 26S proteasome activity in living cells. we divided these cells into ZsGreen-high (low proteasome activity) and ZsGreen-low (high proteasome activity) groups according to their fluorescence intensity. Sphere forming assay revealed that the self-renewal ability of the ZsGreen-high population was significantly higher than that of the ZsGreen-low population. Tumorigenicity assay confirmed that the ZsGreen-high population exhibited higher tumorigenic potential than the ZsGreen-low population. The radioresistance of the ZsGreen-high population of both HeLa and CaSki cells and the chemoresistance of the ZsGreen-high population of CaSki cells were confirmed by clonogenic survival assay and the MTT assay, respectively. These results indicated that ZsGreen-high populations of both the HeLa and CaSki cell lines possess CSC-like properties and therapeutic resistance. We conclude that established cervical cancer cell lines are heterogeneous with regard to their intrinsic proteasome activity and that they contain CSCs-enriched subpopulations that can be identified by low proteasome activity.

Citation Format: Keisuke Tamari, Hideshi Ishii, Kazuhiko Hayashi, Masamitsu Konno, Koichi Kawamoto, Naohiro Nishida, Jun Kozeki, Takahito Fukusumi, Yuichiro Doki, Masaki Mori, Kazuhiko Ogawa. Identification of cervical cancer stem cells by using an imaging system for proteasome activity. [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 3319. doi:10.1158/1538-7445.AM2015-3319
Abstract 3322: Adding pericyte inhibition to combined anti-angiogenesis and irradiation slows tumor growth but increases metastasis in mice model
Authors: Deng Lei , He Jiazhuo , Xue Jianxin , Lan Jie , Zhou Lin , Liu Yongmei , Lu You
Published: August 2015

Increased pericyte coverage has been suggested to be one of the resistant mechanisms of anti-angiogenic treatment (AAT), but targeting pericytes disrupts tumor vascular normalization and abolishes the benefit of oxygenation for the combination of irradiation (IR) with AAT. This raises the question of whether pericyte inhibition should be added to the combination of AAT and IR. We and others have shown that Endostar (Endo), a humanized endostatin, induces a normalization window starting from D5, characterized by decreased tumor hypoxia and increased pericyte coverage of vessels. In this study, we tried to utilize the reduced hypoxia in vascular normalization window to radiosensitize tumor and attempted to test whether inhibiting pericytes after the window may result in optimal anti-tumor effect. Subcutaneous Lewis lung carcinoma was used as the tumor model. Pericyte inhibition was achieved by administering CP673451 (CP) at a dose of 20mg/kg, a platelet-derived growth factor receptor (PDGFR) inhibitor. Endo was given daily for 10 days. 12Gy/1f IR was administered on D5 post-endostar initiation. CP was given from D6 to D10 in order not to interfere with the hypoxia alleviation by Endo when IR was given. We found that percentage of pericyte-covered endothelial cells was the lowest in IR+Endo+CP group compared to IR alone and IR+Endo. IR+Endo+CP significantly reduced tumor growth rate compared to the other two groups. However, the local growth retardation was not translated to mice survival benefits, without even any trend could be suggested. We next found that trimodality treatment resulted in significantly more metastasis in mice lungs. This might be due to increased tumor hypoxia in RT+Endo+CP group, which was found to be the highest among the three. Thus, we hereby conclude that adding pericytes inhibition to the combination of IR and AAT even without interfering vascular normalization when IR is administered confers benefit to local tumor control but not to mice survival, due to increased hypoxia and distant metastasis.

Citation Format: Lei Deng, Jiazhuo He, Jianxin Xue, Jie Lan, Lin Zhou, Yongmei Liu, You Lu. Adding pericyte inhibition to combined anti-angiogenesis and irradiation slows tumor growth but increases metastasis in mice 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 3322. doi:10.1158/1538-7445.AM2015-3322
Abstract 333: Evaluation of the mixed solution of sodium hyaluronate and hydroxyethylstarch on gynecologic cancer cell line
Authors: Lee Ha-Young , Cho Hyun-Jung , Lee Shin-Wha , Lee Sang-Eun , Kim Dae-Yeon , Kim Jong-Hyeok , Kim Yong-Man , Kim Young-Tak , Nam Joo-Hyun
Published: August 2015

Effect of hyaluronic acid and hydroethyl starch mixture in ovarian cancer cell

Objective: Post-operative tissue adhesion is a very serious problem in patients with ovarian cancer. This study is to investigate the role of hyaluronic acid and hydroethyl starch mixture against the tissue adhesion through in vitro experimentations with ovarian cancer cell lines.

Methods: We used different concentrations of hyaluronic acid and hydroethyl starch (HA/HES) mixture in SKOV3 cell lines. We performed the adhesion and proliferation inhibition assay in different method, mixing and spreading cells. For the migration inhibition assay, we evaluated the effect according to the concentration of HA/HES mixture. The wound-healing assay was performed after artificial injury and physical barrier of 100% HA/HES mixture was evaluated.

Results: In adhesion and proliferation inhibition assay, the viable adherent cells were decreased according to the concentration of HA/HES mixture. The confluence increased over time, except for 100% HA/HES mixture. This result was more obvious in spreading method. The wound recovery was slow in high concentration of HA/HES mixture in the migration inhibition assay and wound-healing assay. The physical barrier of 100% HA/HES mixture inhibited the cell growth until 48 hours.

Conclusion: The proliferation and recovery inhibition of SKOV3 cell lines using HA/HES mixture was confirmed and the inhibitory effect appeared to be in proportion of the concentration of HA/HES mixture. This growth inhibition effect of HA/HES mixture will be useful clinically in patients with ovarian cancer

Citation Format: Ha-Young Lee, Hyun-Jung Cho, Shin-Wha Lee, Sang-Eun Lee, Dae-Yeon Kim, Jong-Hyeok Kim, Yong-Man Kim, Young-Tak Kim, Joo-Hyun Nam. Evaluation of the mixed solution of sodium hyaluronate and hydroxyethylstarch on gynecologic cancer cell line. [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 333. doi:10.1158/1538-7445.AM2015-333
Abstract 3333: The expression of FKTN gene in gastric cancer related to radiation exposure: research on atomic bomb survivors
Authors: Pham Trang T.B.
Published: August 2015

While radiation use increases, the Life Span Study (LSS) atomic bomb survivor cohort of Radiation Effects Reserach Foundation (RERF) from 1951-2003 shows a significant cancer risk for most major organs in the body, including stomach. To identify genes associated with gastric cancer, we utilize Escherichia coli ampicillin secretion trap (CAST) method, which is a powerful tool for novel cancer biomarkers. By this method, we found that FKTN (fukutin) was overexpressed in gastric cancer. Fukutin localizes to the Golgi apparatus and normally has low expression, but overexpresses in some cell lines. It is thought to forms a complex with protein POMGnT1, a glycosyltransferase involved in the synthesis of O-mannosyl glycan attached to mucin-like domain of alpha-dystroglycan. The POMGnT1 activity was also downregulated by fukutin mutation and resulted in Fukuyama congenital muscular dystrophy: brain malformation, one of the most common autosomal-recessive disorders in Japan. To elucidate the molecular mechanisms of radiation carcinogenesis, we analyzed FKTN expression from sporadic gastric cancer cohort and LSS cohort by immunohistochemistry. In the sporadic cohort of 695 gastric cancer samples, there was a significant correlation of FKTN expression in differentiated type gastric cancer. For mucin markers, a significant relationship was detected between FKTN and CD10 (p<0.0001). The LSS cohort of 86 gastric cancer samples showed FTKN expression downregulated in high-dose than in low-dose exposed patients (p<0.0001), but with no correlation between FKTN and mucin phenotype expression. This is the first analysis on the expression of fukutin protein in gastric cancers among atomic-bomb survivors and the radiation dose response, using immunohistochemistry. These results suggest that FTKN has a possible relationship with radiation exposure and gastric cancer among atomic bomb survivors.

Citation Format: Trang T.B. Pham. The expression of FKTN gene in gastric cancer related to radiation exposure: research on atomic bomb survivors. [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 3333. doi:10.1158/1538-7445.AM2015-3333
Abstract 3335: Targeting Egr1 selectively radioprotects normal tissues while killing cancer cells
Authors: Zhao Diana Yi , Jacobs Keith M. , Hallahan Dennis E. , Thotala Dinesh
Published: August 2015

Radiotherapy is a common therapeutic modality for a wide range of malignancies. Normal tissue toxicity reduces the therapeutic index of radiotherapy and decreases the quality of life for cancer survivors. Apoptosis is a key element of the radiation response in normal tissues like the hippocampus and small intestine, and results in neurocognitive disorders and intestinal malabsorption. The Early Growth Response 1 (Egr1) transcription factor mediates radiation-induced apoptosis since it activates the transcription of pro-apoptosis genes in response to ionizing radiation (IR). Therefore, we hypothesized that the genetic abrogation of Egr1 and the pharmacological inhibition of its transcriptional activity could attenuate radiation-induced apoptosis in normal tissues. We demonstrated that Egr1 null mice had less apoptosis in the hippocampus and intestine following irradiation as compared to their wild-type littermates. Alternatively, mice treated with Mithramycin A, a drug that prevents binding of Egr1 to target promoters, also displayed reduced apoptosis following irradiation in the intestine as compared to mice treated with vehicle. In cell culture based studies, shRNA-mediated silencing of Egr1 expression dampened apoptosis and enhanced the clonogenic survival of irradiated HT22 hippocampal neuronal cells and IEC6 intestinal epithelial cells. Mechanistically, these events involved an abrogation of p53 induction by IR and an increase in the ratio of Bcl-2/Bax expression. In contrast, targeted silencing of Egr1 in two cancer cell lines (GL261 glioma cells, HCT116 colorectal cancer cells) was not radioprotective, since it reduced their growth while also sensitizing them to radiation-induced death. These results strongly support the potential of targeted silencing of Egr1 in order to improve the therapeutic index of radiotherapy and minimize the normal tissue complications associated with radiation therapy.

Note: This abstract was not presented at the meeting.

Citation Format: Diana Yi Zhao, Keith M. Jacobs, Dennis E. Hallahan, Dinesh Thotala. Targeting Egr1 selectively radioprotects normal tissues while killing cancer cells. [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 3335. doi:10.1158/1538-7445.AM2015-3335
Abstract 334: Glucose regulates chemoresistance and stemness via Wnt/β-catenin signaling in ovarian cancer cells
Authors: Gwak HyeRan , Kim Soochi , Jo UnTek , Song Yong Sang
Published: August 2015

Glucose metabolic alteration is one of the hallmarks of ovarian cancer. Here, we first demonstrated the function of glucose as a signature molecule that regulated chemosensitivities in ovarian cancer cells. To verify the role of glucose on the chemoresistance in ovarian cancer cells, we used various concentrations of glucose containing media (0 mM to 20 mM). Glucose reduction inhibited proliferation of various ovarian cancer cell types; irrespective of p53 status and cisplatin (CDDP) sensitivities. Interestingly, CDDP (10 to 20 μM) induced apoptosis were increased and expressions of p-glycoprotein (multidrug resistance protein 1, MDR1) and ABCG2 (ATP-binding cassette sub-family G member 2) were decreased in both mRNA and protein levels by glucose free conditions. Moreover, sphere forming capacities and stemness related gene expressions, SOX2, OCT4, BMI1 and Nanog, were also decreased in ovarian cancer cells in response to glucose starvation. Suppression of CDDP resistance and stemness in ovarian cancer was found to be associated with the activation of GSK3β in response to limitation of glucose availabilities, as confirmed by the function of the GSK3β inhibitors (LiCl and SB SB216763) and also downstream molecules, β-catenin and cyclin D1. In contrast, high glucose (20 mM) environment enhanced CDDP resistance in ovarian cancer cells via elevation of p-glycoprotein and ABCG2 expressions, sphere forming capacities and stemness related gene expressions. Moreover, CDDP (10 to 20 μM) in high concentration of glucose media induced apoptotic cell death were significantly decreased. This study described the novel function of glucose in chemoresistance in ovarian cancer cells by deregulation of multidrug efflux pumps and stemness through Wnt/ β-catenin signaling cascades.

Citation Format: HyeRan Gwak, Soochi Kim, UnTek Jo, Yong Sang Song. Glucose regulates chemoresistance and stemness via Wnt/β-catenin signaling in ovarian cancer cells. [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 334. doi:10.1158/1538-7445.AM2015-334
Abstract 3341: TP508 sensitizes cancer stem cells to radiation and delays brain cancer cell relapse in vitro
Authors: Moya Stephanie M. , Kantara Carla , Sower Laurie , Carney Darrell H.
Published: August 2015

In 2014, approximately 70,000 adults and 4,500 children are expected to be diagnosed with brain cancer in the U.S alone, making this disease the second leading cause of cancer related deaths in children and young adults. Currently available therapies such as radio/chemotherapy are ineffective in completely eradicating the tumor bulk and cancer stem cells within the tumor, thus allowing recurrence of the disease. Therefore, it is important to develop novel therapeutics that sensitize cancer cells to make them more susceptible to radiation therapy and reduce the number of viable cancer stem cells. Preliminary data showed that injection of TP508, a 23 amino acid thrombin derived peptide drug, increased brain tumor shrinkage post-radiation therapy (RT) in mice. To study this phenomenon, we chose to use an in vitro model of tumor relapse that selects for and allows quantification of tumor stem cells in 3D spheroid cultures. Our goal was to determine whether TP508 could specifically sensitize cancer stem cells to the effects of RT and prevent or delay stem cell mediated in vitro tumor relapse. Medulloblastoma (Daoy) cells were grown as 3D spheroids in vitro to select for the growth of stem cells. Primary spheroids were treated on day 5 with either saline or TP508 (0.5mg/mL), and exposed to radiation (10Gy) on day 7 using a Cs137 source. One day post-RT, cells were dissociated and cultured as secondary spheroids to determine the stemness or relapse potential of the cells following RT. Daily images were taken to quantify neurosphere formation. Spheroids were also analyzed for viability, apoptosis, proliferation (PCNA), and for stem cell markers (CD133/LGR5/CD44/DCLK1/Sox-2), using western blots, flow cytometry, and immuno-fluorescent staining. TP508 treatment of Daoy cell spheroids prior to RT significantly delayed the formation of secondary spheroids, decreased cell viability, number of proliferating cells, and the number of cells expressing cancer stem cell markers. Thus, in this in vitro assay, TP508 sensitized cancer stem cells to the effects of RT and delayed relapse of brain tumor stem cells in vitro. These findings suggest that injection of TP508 prior to RT treatment may sensitize cancer stem cells to radiation and help prevent brain tumor relapse. Studies are underway to determine the mechanism by which TP508 sensitizes cancer stem cells to RT and whether TP508 has similar effects on other types of cancer stem cells. This work was supported by NCI Contract # HHSN261201300076C to DC.

Citation Format: Stephanie M. Moya, Carla Kantara, Laurie Sower, Darrell H. Carney. TP508 sensitizes cancer stem cells to radiation and delays brain cancer cell relapse in vitro. [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 3341. doi:10.1158/1538-7445.AM2015-3341
Abstract 3360: Macrophages promote resistance to pegylated arginine deiminase in malignant pleural mesothelioma
Authors: Phillips Melissa M. , Khadeir Ramsay , Tookman Laura , McCarthy Fiona , Steele Jeremy , Bomalaski John , Ghazaly Essam , Szlosarek Peter W.
Published: August 2015

Background

Approximately 50% of all malignant pleural mesotheliomas (MPM) are deficient in argininosuccinate synthetase (ASS1), the rate-limiting enzyme in arginine biosynthesis, and are therefore sensitive to arginine deprivation. This discovery in MPM has been translated into the clinic using the arginine depletor pegylated arginine deiminase (ADI-PEG20) which showed a halving in the risk of disease progression in a randomized phase II study (Szlosarek et al, ASCO 2014). However, unstudied to date, stromal resistance to ADI-PEG20 may reduce its efficacy. Here, we studied the effect of macrophages, which are abundant in mesothelioma, on the tumor cytotoxicity of ADI-PEG20.

Methods

ASS1 negative MPM cell lines treated with ADI-PEG20 were analysed using the Affymetrix Human Genome U133 Plus 2.0 array, with validation of genes involved in stromal-tumor cell communication. Additional studies involved using tumor-macrophage co-culture experiments, mass spectrometry and an MPM xenograft model.

Results

A distinct pro-inflammatory cytokine gene expression signature involved in macrophage recruitment and activation was identified in the ADI-PEG20-treated ASS1 negative MPM cell lines. Notably, a significant increase in ASS1 negative MPM cell viability was seen upon co-culture with macrophages in the presence of ADI-PEG20. This was accompanied by a significant increase in ASS1 expression in co-cultured macrophages, with a corresponding increase in argininosuccinate lyase (ASL) expression in co-cultured tumor cells and a doubling in levels of the arginine precursor, argininosuccinate, in cell supernatant. The addition of argininosuccinate to tumor cell media rescued ASS1 negative MPM cells from ADI-PEG20 cytotoxicity, while the macrophage-mediated resistance to ADI-PEG20 was abrogated following ASL knockdown in MPM cells. Finally, xenograft studies demonstrated a significant reduction in tumor volume in mice treated with ADI-PEG20 in combination with macrophage depletion, compared with ADI-PEG20 monotherapy.

Conclusion

Collectively, our data indicate that as a result of metabolic ‘cross-talk’ between macrophages and ASS1 negative MPM cells, macrophages mediate MPM resistance to ADI-PEG20 via the provision of argininosuccinate. Our studies provide a rationale for combining ADI-PEG20 with an inhibitor of macrophage recruitment in the treatment of ASS1-deficient mesothelioma.

Citation Format: Melissa M. Phillips, Ramsay Khadeir, Laura Tookman, Fiona McCarthy, Jeremy Steele, John Bomalaski, Essam Ghazaly, Peter W. Szlosarek. Macrophages promote resistance to pegylated arginine deiminase in malignant pleural mesothelioma. [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 3360. doi:10.1158/1538-7445.AM2015-3360
Abstract 3363: RAGE/S100A7/Stat3-axis enhances breast cancer growth and metastasis via modulating tumor microenvironment
Authors: Nasser Mohd W. , Wani Nissar A. , Ravi Janani , Amponsah Grace A. , Ahirwar Dinesh K. , Powell Catherine A. , Elbaz Mohamad , Zhao Helong , Shilo Konstantin , Ganju Ramesh K.
Published: August 2015

The Receptor for Advanced Glycation Endproducts (RAGE), a multifunctional, multi-ligand receptor, has been shown to play an important role in inflammation. However, not much is known about its role in breast cancer growth and metastasis. In this report, we observed that RAGE expression is upregulated in triple negative breast cancer (TNBC) cell lines, primary tumors and lymph-node metastasis samples. RAGE-/- mice show reduced breast cancer growth. Additionally, blocking RAGE with neutralizing-antibody inhibited lung metastasis in an intracardiac mouse model. Further elucidation of RAGE-mediated mechanisms revealed that RAGE binds to S100A7 and mediates S100A7-induced cell migration, Stat3 and NF-ĸB activation. Our results also indicate that S100A7/RAGE axis-modulates invasion/migration through Stat3 dependent MMP9 activation. In addition, RAGE neutralizing antibody and soluble RAGE inhibited breast cancer progression and metastasis in the inducible mS100a7a15 mouse model system. We demonstrated that RAGE/S100A7 enhanced mammary hyperplasia, tumor growth and metastasis through Stat3 activation. Notably, our studies revealed that RAGE/mS100a7a15 modulates the breast tumor microenvironment through recruitment of phospho-Stat3/MMP9-positive tumor-associated macrophages. Our studies suggest that RAGE expression could be used as a novel biomarker for aggressive/invasive breast cancer, especially TNBC. Collectively, these findings suggest that RAGE/S100A7/Stat3-axis has a novel role in linking inflammation to the development of invasive/aggressive breast cancer including TNBC.

Citation Format: Mohd W. Nasser, Nissar A. Wani, Janani Ravi, Grace A. Amponsah, Dinesh K. Ahirwar, Catherine A. Powell, Mohamad Elbaz, Helong Zhao, Konstantin Shilo, Ramesh K. Ganju. RAGE/S100A7/Stat3-axis enhances breast cancer growth and metastasis via modulating tumor microenvironment. [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 3363. doi:10.1158/1538-7445.AM2015-3363
Abstract 3375: Can prostate cancer derived neuromodulators regulate intratumoral axonogenesis
Authors: Gutkin Dmitriy W. , Shurin Galina V. , Shurin Michael R. , Keskinov Anton A.
Published: August 2015

Well-developed angiolymphatic network in solid malignant tumors has been proven as an important requirement for tumor growth and progression. However, the level of innervation of the tumor mass is not a well-known factor in tumor development and spreading. Several mechanisms may be responsible for activation and inhibition of neural elements in the tumor microenvironment, including numerous neurotrophic factors (NF) that have axon attractive and axon repellent properties. We have examined 40 cases of human prostate adenocarcinoma by immunohistochemical stains, highlighting neural elements (S-100, PGP9.5, neurofilament) and blood vessels (CD31, CD34). The result revealed that while blood vessels were abundant in the tumor tissue, the neural structures were very scarce compared to benign prostate tissue. Next, in a model system, cell-free supernatants from TRAMP C2 and RM-1murine prostate cancer cell lines were added to the murine neuron cultures, and morphology of the neurons was analyzed by assessing the number of cells with neurites, the number of neuritis per cell, and the lengths of neurites. The data showed that tumor-conditioned media of TRAMP-C2 and RM-1 cells significantly inhibit the neurite growth in mouse neuron cultures. Analysis of gene expression in murine prostatic adenocarcinoma by RT-PCR revealed expression of Semaphorin 4A, Semaphorin 3F, Ephrin A1, Ephrin B2, and Slit2. Finally, the transfection of murine prostate cancer cells with Ephrin 1A and Slit2 siRNAs has prevented their neuron-inhibiting effect. Together, our results suggest that prostate carcinoma cells express NF factors that may inhibit neurite growth and lead to down-regulated innervation of tumor mass. The role of this phenomenon in tumor growth and metastatic potential is under investigation.

Citation Format: Dmitriy W. Gutkin, Galina V. Shurin, Michael R. Shurin, Anton A. Keskinov. Can prostate cancer derived neuromodulators regulate intratumoral axonogenesis. [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 3375. doi:10.1158/1538-7445.AM2015-3375
Abstract 340: Identification of p62/IMP2 novel targets in breast cancer metastasis
Authors: Li Yang , Peng Bo , Lei Ningjing , Qian Wei , Francia Giulio , Zhang Jianying
Published: August 2015

As one of the most common types of cancer in women, breast cancer is far from being well controlled, especially when the cancer spreads to other organs in the body. Therefore, it is important to understand the molecular and cellular mechanism of metastatic breast cancer, and to identify new proteins that regulate the metastatic process, as such proteins may be used as targets for possible therapeutic intervention. Our previous study showed that p62/IMP2 (IGF2 mRNA binding protein 2) is overexpressed in breast cancer tissues. In our present study, some variants were generated with stable overexpression of p62/IMP2 in MDA-MB-231 cells and LM2-4 cells, and we found that overexpression of p62/IMP2 can increase breast cancer cell migration and reduce cell adhesion. We also found that the p62/IMP2 can bind to c-myc mRNA and increase the expression of c-myc in the two cell lines. To identify new p62/IMP2 targets related with cell adhesion, we performed a Human Extracelluar Matrix & Adhesion Molecules PCR Array to see which gene expression changes in the p62/IMP2 overexpressed cells, compared to the control cells. Our results showed that 18 adhesion molecules are up-regulated with the overexpression of p62/IMP2 (Fold regulation>2, p<0.05) while 2 are down-regulated. We selected 7 up-regulated genes (such as CTGF, HTBS1 and COL15A1) and a down-regulated gene (ITGB4) as candidates for the further evaluation by real time PCR. All candidates genes were further verified byRNA immunoprecipitation (RIP). The data from RIP suggested that connective tissue growth factor (CTGF) may be a potential target of p62/IMP2. These results provided evidences for a better understanding of breast cancer metastasis and targets for future anti-metastatic drug design.

Note: This abstract was not presented at the meeting.

Citation Format: Yang Li, Bo Peng, Ningjing Lei, Wei Qian, Giulio Francia, Jianying Zhang. Identification of p62/IMP2 novel targets in breast cancer 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 340. doi:10.1158/1538-7445.AM2015-340
Abstract 3429: Patient awareness of staging in cancer diagnosis: Helpful or hurtful
Authors: Schaap Ariel , Aggarwal Rashi , Domogauer Jason
Published: August 2015

Objective:

In this study we will examine if education levels of English-speaking patients in New Jersey Medical School's Outpatient Cancer Center is correlated with awareness of their disease. Next we will examine their level of depression, as well as explore if patients’ awareness of their disease is correlated with depression. Lastly we will explore if patients’ awareness of their disease is correlated with their level of self-perceived stigma regarding depression.

Methods:

This project is part of a larger questionnaire-based study examining depression prevalence and its associated stigma in cancer patients. Participants consisted of English-speaking oncology patients at New Jersey Medical Cancer Center outpatient oncology clinics. The parts of the questionnaire that I analyzed for this study:

1. What is your level of education?

2. What stage was your cancer in when you were diagnosed?

3. Currently is your cancer localized to a single area or has it metastasized to other areas?

4. Depression symptoms were assessed using the CES-D depression scale.

5. Self-stigma was assessed using Depression Stigma Scale.

Summary:

An increased level of education was associated with a significantly increased level of awareness of cancer staging, linear by linear association (1, N = 223) = 9.353 p = .002. 31% of patients did not know their cancer staging and/or whether or not it had metastasized. Out of the people who were aware of staging only 28.5% were depressed, whereas 34.8% of people who were unaware of staging and/or metastasis were depressed, Fisher Exact test p = .308. While this is not significant, the trend is an important one to be aware of and to examine in future research.

Patients who were aware of their staging and metastasis had a lower mean self-perceived stigma than patients who were unaware of staging and/or metastasis, 14.87 and 15.19 respectively, t(216) = .32, p = .031

Conclusion:

Doctors must recognize that patients in low educated demographic areas, such as Newark, require special attention and need to be evaluated on whether or not they understood what the doctor explained to them about their disease. Further research should focus on analyzing the exact reasons underlying this lack of patient knowledge. In addition, the correlation between lack of knowledge and depression should continue to be explored as patient control and autonomy can improve patient outcomes. Doctors should recognize that patients may have a high self-perceived stigma associated with depression, which can act as a barrier to treatment. Future studies should look at how to dispel that stigma.

Citation Format: Ariel Schaap, Rashi Aggarwal, Jason Domogauer. Patient awareness of staging in cancer diagnosis: Helpful or hurtful. [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 3429. doi:10.1158/1538-7445.AM2015-3429
Abstract 3440: High-dose-rate brachytherapy as monotherapy for favorable-risk adenocarcinoma of the prostate delivered in a single 19 Gy fraction: Preliminary results of a prospective pilot study
Authors: Dahlbeck Scott , Hansen Chase C. , deRiese Werner , Kagan A. Robert , Torres Carlos , Chiriva-Internati Maurizio , Cobos Everardo , Figueroa Jose A. , Nguyen Diane , Tijani Lukman , Evans Jaden D.
Published: August 2015

INTRODUCTION: We evaluated the acute genitourinary (GU) and gastrointestinal (GI) toxicities, health-related quality of life (HRQOL) factors, biochemical control rates, and technical feasibility of high-dose-rate (HDR) brachytherapy as monotherapy for favorable-risk prostate cancer delivered in a single 19 Gy fraction.

METHODS: A single-institution, prospective pilot study was performed by evaluating 6 patients with low- and intermediate-risk prostate cancer treated with high-dose-rate (HDR) brachytherapy as monotherapy. Patients received a single 19 Gy fraction as HDR monotherapy without the use of a transperineal hyaluronic acid injection. Patients were assessed according to the Common Terminology Criteria for Adverse Events version 4.0 and Health-Related Quality of Life (HRQOL) questionnaires. Additionally, prostate specific antigen levels have been followed for evidence of biochemical failure.

RESULTS: All 6 patients tolerated the implant well and were all discharged home the same day. Median follow-up was 15 months with all subjects followed for at least 12 months. No grade 3, 4, or 5 toxicities were observed. Two of the 6 patients experienced grade 2 GU toxicity. One patient experienced grade 2 GI toxicity. HRQOL bowel and urinary assessments revealed peak complaints at 3 months which returned to baseline at 6 months. There have been no biochemical relapses.

CONCLUSION: This is the first study using HDR brachytherapy as monotherapy for favorable-risk prostate cancer using one implant delivered in a single 19 Gy dose in the United States. All patients demonstrated acceptable acute toxicities and were pleased with their cost-effective treatment choice.

Citation Format: Scott Dahlbeck, Chase C. Hansen, Werner deRiese, A. Robert Kagan, Carlos Torres, Maurizio Chiriva-Internati, Everardo Cobos, Jose A. Figueroa, Diane Nguyen, Lukman Tijani, Jaden D. Evans. High-dose-rate brachytherapy as monotherapy for favorable-risk adenocarcinoma of the prostate delivered in a single 19 Gy fraction: Preliminary results of a prospective pilot study. [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 3440. doi:10.1158/1538-7445.AM2015-3440
Abstract 3462: Proteomic profiling reveals novel targets for combination treatment with lanreotide in neuroendocrine tumors
Authors: Fotouhi Omid , Kjellin Hanna , Hashemi Jamileh , Lu Ming , Juhlin Christofer , Höög Anders , Zedenius Jan , Larsson Catharina , Lehtiö Janne , Kjellman Magnus
Published: August 2015

Somatostatin analogues (SSAs) are well established in the treatment of neuroendocrine tumors (NETs), including small intestinal NET (SI-NET) due to their tumor growth arrest and symptom relieving characteristics. Here we determined the direct effects of lanreotide treatment on NET cell lines and a primary SI-NET culture. The cell lines HC45 and H727 were treated with a pharmacological concentration of 10 nM lanreotide for different time periods and the proteome, analyzed by in-depth HiRIEF LC-MS/MS, was compared with the proteome of non-treated cells. We quantified 6 451 and 7 801 proteins in HC45 and H727. Differential expression of the candidate proteins APC, BIRC5/survivin, BMPER, C14orf142, FYN, INSM1 and SPAG16 was confirmed by Western blotting. The possible direct anti-proliferative effect of lanreotide was evaluated in the primary SI-NET culture and the HC45, H727 and BON1 cell lines, which showed only a slight inhibition of proliferation. However, based on proteomics driven pathway analysis a significant synergy in anti-proliferative effect of lanreotide was observed when the small molecule IGF1R inhibitor, NVP-AEW541, was combined with lanreotide .This effective combination also represents an indirect model of suppressed IGF1 activity in the NET patients treated with SSA. Targeting of survivin with the small molecule YM155 dramatically reduced the proliferation of HC45. We also studied the effect of lanreotide alone or in combination with IGF-1R inhibitor on signaling pathways PI3K/Akt, Erk1/2 and P38. P38, Erk1/2 and GSK3β phosphorylation was equally increased by lanreotide and/or NVP-AEW541, however, lanreotide-induced AktS473 dephosphorylation was reversed after6 hours, unlike consistent AktS473 dephosphorylation after treating with NVP-AEW451 alone or in combination. In conclusion, we report growth inhibitory and potential feedback pro-growth signaling of the SSA lanreotide on NET cells, highlighting the role of SSAs in modulation of IGF1 in NET cell growth arrest and propose combination treatment candidate targets for improved efficacy of this agent.

Citation Format: Omid Fotouhi, Hanna Kjellin, Jamileh Hashemi, Ming Lu, Christofer Juhlin, Anders Höög, Jan Zedenius, Catharina Larsson, Janne Lehtiö, Magnus Kjellman. Proteomic profiling reveals novel targets for combination treatment with lanreotide in neuroendocrine tumors. [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 3462. doi:10.1158/1538-7445.AM2015-3462
Abstract 3485: Uncertainty envelope to evaluate drug interactivity with statistical analysis
Authors: Zhao Liang , Au Jessie L.-s. , Wientjes M. Guill
Published: August 2015

Introduction. Development of new therapies for cancer and infections frequently involves combinations of two or more drugs. Existing methods to quantify drug interactivity are suitable for analyzing drugs with parallel concentration-effect curves and similar maximum drug effects. This study established a method for the remaining situations.

Methods. Equations describing zero-interactivity effects of combination therapy were developed based on the principle that a drug cannot interact with itself (i.e., additivity only). These equations were used to convert the concentration-effect (C-E) curve of the combination to C-E curve for each of the two drugs in the combination as if they were used as single agents (single agent-equivalent curves). Confidence intervals (CI) of these curves were generated using the error propagation method. Effect data of drug combinations were analyzed by the new and existing methods.

Results. The calculated single agent-equivalent curves plus 95% CI defined the boundaries of an envelope for additive drug effects (Uncertainty Envelope). The Envelope shape and size were determined by differences in the shapes of single agent C-E curves. The nature of drug interactivity is indicated by the location of experimental data points of the combination; data points located within the Envelope indicate additivity, whereas points above and below the Envelope respectively indicate statistically significant synergy and antagonism. In comparison with the existing methods, the Envelope method yielded more conservative results (additivity vs. synergy or antagonism).

Conclusion. Uncertainty Envelope method enables the interactivity analysis between drugs with nonparallel C-E curves and unequal maximal effect, with statistical confidence.

Citation Format: Liang Zhao, Jessie L.-s. Au, M. Guill Wientjes. Uncertainty envelope to evaluate drug interactivity with statistical analysis. [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 3485. doi:10.1158/1538-7445.AM2015-3485
Abstract 3493: Disulfiram a dual mgmt and aldehyde dehydrogenase inhibitor sensitizes pancreatic cancer to gemcitabine and abraxane
Authors: Bobustuc George C. , Holmuhamedov Alisher , Srivenugopal Kalkunte S. , Frick Jacob C. , WEESE JAMES L. , Konduri Santhi D.
Published: August 2015

Background: O6 methylguanine DNA methyltransferase (MGMT) is overexpressed in a majority of cancers, including pancreatic cancer. MGMT has been the focus of significant research for its role in the repair of DNA damage caused by chemotherapeutic agents. Aldehyde Dehydrogenase as a progenitor/stem cell marker has been linked to chemotherapy and radiation resistance.

Methods: We tested the administration of Antabuse (disulfiram, DSF) at nontoxic doses in combination with gemcitabine (GEM) and Abraxane (ABX) on pancreatic cancer cells.

Results: DSF at very low doses (achievable in human serum with standard DSF clinical dosing) decreases pancreatic cancer cell growth in a dose-dependent manner. DSF further sensitizes pancreatic cancer cells to GEM and ABX and significantly inhibits pancreatic cancer growth without causing unwanted side effects to the normal pancreatic cells. Dose effect and isobologram studies confirm synergistic activity of DSF and ABX combination and moderate synergism for DSF and GEM combination. DSF, either alone or in combination with GEM (DSF ± GEM) and/or ABX (DSF ± ABX), significantly inhibits expression of MGMT, aldehyde dehydrogenase, antigen Ki-67 and BIRC5 gene (survivin) - all involved in pancreatic cancer tumorigenesis. DSF+/-GEM+/-ABX induces the expression of p21Cip1, a cell cycle inhibitor. We show an inverse correlation between aldehyde dehydrogenase activity and MGMT expression, where MGMT expression is high, aldehyde dehydrogenase activity is low and where aldehyde dehydrogenase activity is high, MGMT levels were low. The ALDH3A1 expression is significantly higher in highly metastatic cancer cells (L3.6pl) compared to other metastatic cancer cells (PANC1, Mia PaCa2, ASPC1). We further report that in our model DSF at very low doses (achievable in human serum with standard DSF clinical dosing) effectively inhibits both aldehyde dehydrogenase and MGMT in pancreatic cancer cells. DSF also induces caspase activation indicative of mitochondrial induced apoptosis in these cells.

Conclusions: Our findings suggest that DSF as dual inhibitor of MGMT and aldehyde dehydrogenase potentially provides a novel therapeutic approach to inhibiting pancreatic cancer growth/metastasis and synergistically enhances GEM and ABX activity.

Citation Format: George C. Bobustuc, Alisher Holmuhamedov, Kalkunte S. Srivenugopal, Jacob C. Frick, JAMES L. WEESE, Santhi D. Konduri. Disulfiram a dual mgmt and aldehyde dehydrogenase inhibitor sensitizes pancreatic cancer to gemcitabine and abraxane. [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 3493. doi:10.1158/1538-7445.AM2015-3493