bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4618: Elucidating the role of distinct glutaminase isoforms in cancer.
Authors: Liu Bonnie , Song Kyung , Kwong Mandy , Gao Min , Hong Rebecca , Nannini Michelle , Sampath Deepak , Belvin Marcia , Peterson David , Evangelista Marie , Daemen Anneleen , Firestein Ron , Hatzivassiliou Georgia
Published: August 2014

Glutaminase is a key enzyme for the conversion of glutamine to glutamate with subsequent use as a fuel in the TCA cycle, glutathione production and protein synthesis. A subset of tumor cells shows preferential dependency on glutamine and glutaminase for cell proliferation and survival, consistent with “metabolic addiction” to this pathway. Between two glutaminase loci in the genome, the gene for kidney-type glutaminase (Gls1) has 2 alternatively spliced products (KGA and GAC), with distinct regulation and expression from liver-type glutaminase (Gls2). Furthermore, based on distinct C-termini and enzymatic properties, it has been proposed that the Gls1 alternatively spliced isoforms (KGA and GAC) may themselves have distinct functions. Our data further elucidates the role of glutaminase in cancer by addressing the following key questions: a) what is the major role of glutaminase in cells most sensitive to its inhibition? b) is GAC interchangeable with KGA in its expression pattern and role in tumor cells? c) how do tumors adapt to glutaminase inhibition? By generating and using a panel of GAC and KGA isoform-selective reagents we show that the two isoforms have distinct patterns of expression and distinct roles in cell proliferation, migration and tumor growth in vivo. In addition, based on metabolomics, functional and complementation assays, we propose a mechanistic basis for tumor cell dependency on glutaminase and the tumor adaptive response to glutaminase inhibition. Our data reveal the basis for the addiction of tumor subsets to glutaminase and elucidate the role of distinct isoforms of kidney-type glutaminase isoforms in cancer.

Citation Format: Bonnie Liu, Kyung Song, Mandy Kwong, Min Gao, Rebecca Hong, Michelle Nannini, Deepak Sampath, Marcia Belvin, David Peterson, Marie Evangelista, Anneleen Daemen, Ron Firestein, Georgia Hatzivassiliou. Elucidating the role of distinct glutaminase isoforms in cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4618. doi:10.1158/1538-7445.AM2013-4618
Abstract 4652: Proteomic profiling of platinum based anticancer drugs.
Authors: Dzubak Petr , Ozdian Tomas , Holub Dusan , Rylova Gabriela , Rehulka Jiri , kollareddy Madhusudhan , Mlcuchova Daniela , Hajduch Marian
Published: August 2014

Carboplatin, cisplatin and oxaliplatin are structurally similar anticancer drugs and their mechanism of action is permanent coordination to DNA and interfering with DNA repair. However, these drugs have different anticancer activities in human tumors, adverse effects and dose limiting toxicities. Therefore, we performed complex proteomic profiling in order to characterize cellular effects of all three platinum based anticancer drugs. To identify the molecular basis of different pharmacological properties, expressional characteristics of cells exposed to vincristine and daunorubicin were used in parallel as prototypes of drugs with different mechanism of action. T-lymphoblastic leukemia CCRF-CEM cells were treated with 5x IC50 concentrations of individual drugs for time equivalent to half-time to induction apoptosis. Untreated (control) cells were cultured with heavy amino acid labeled media in order to enable relative quantification of peptide expression using SILAC technology, LC-MS/MS and LC-MALDI-TOF/TOF. Results were processed by Proteinscape software equipped by Mascot and Phenyx analytical algorithms. We identified 646 ± 133 proteins by ESI and 312 ± 85 proteins by MALDI in average of all fifteen replicates. Identified results were further processed by in-house written script to remove outlier peptides and to normalize individual runs. All replicates were summed, and statistic analysis was done. To see up- and down- regulated proteins, the hierarchical clustering and PCA analysis was performed. Hierarchical clustering revealed that cisplatin and carboplatin shares the same cluster what is conclusive with literature. Surprisingly, the cluster of daunorubicin and vincristine was close to cisplatin and carboplatin and the oxaliplatin was in the separated cluster from rest of four drugs. PCA analysis revealed 30 proteins with contribution higher than 1 %. The highest contribution (7.5 %) to drug clustering have proteins involved in DNA remodeling. These proteins could be used as specific biomarkers of therapy by platinum based compounds.

Citation Format: Petr Dzubak, Tomas Ozdian, Dusan Holub, Gabriela Rylova, Jiri Rehulka, Madhusudhan kollareddy, Daniela Mlcuchova, Marian Hajduch. Proteomic profiling of platinum based anticancer drugs. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4652. doi:10.1158/1538-7445.AM2013-4652
Abstract 4849: An association between genetic variant in Epidermal Growth Factor (EGF) gene and survival benefit from epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) in never smokers with advanced adenocarcinoma of the lung.
Authors: Lee Youngjoo , Yoon Kyong-Ah , Lee Jin Soo
Published: August 2014

Background

EGFR mutations are strong predictors for efficacy to EGFR-TKIs in non-small cell lung cancer (NSCLC). However, it is unknown whether inherited genetic variants can influence the EGFR-TKI efficacy in patients harboring EGFR mutations.

Methods

We genotyped 271,817 single-nucleotide polymorphisms (SNP) in 348 advanced NSCLC patients who received chemotherapy. Seventeen top survival-related SNPs were identified using cut-off minimum P-value of <5.0 x 10−5 in at least 70% of 1000 bootstrap samples. Among the 17 genes, we focused on whether SNPs in the EGF gene have predictive value for survival benefit from EGFR-TKIs in 215 never smokers with advanced adenocarcinoma of the lung, which were selected as EGFR mutation-enriched patients, receiving EGFR-TKI treatment during the disease course.

Results

The survival benefit by treatment with EGFR-TKIs was different between patients with common and variant genotypes of rs11098063 at the 4q25: EGF gene [hazard ratio (HR) of ever to never use of EGFR-TKIs, 0.72 (95% CI, 0.42-1.22) in CC genotype vs. 0.27 (95% CI, 0.15-0.50) in CT/TT genotype; interaction of SNP*EGFR-TKIs, P = 0.075] in Cox proportional hazard regression model adjusted for known prognostic factors. In the CT/TT group (n= 89), there was a significant improvement in survival with EGFR-TKIs [never vs. ever use of EGFR-TKIs; median overall survival (OS), 16 (95% CI, 12-21) vs. 37 (95% CI, 31-42) months; log-rank test, P < 0.001]. However, the CC group (n= 126) did not show a significant difference in survival according to use of EGFR-TKIs [never vs. ever use of EGFR-TKIs; median OS, 14 (95% CI, 9-18) vs. 25 (95% CI, 23-27) months; log-rank test, P = 0.082]. On the other hand, the SNP of the EGF gene was not associated with response rate or progression-free survival of EGFR-TKIs.

Conclusions

These results suggested the genetic variants at the EGF gene might contribute to the survival difference associated with EGFR-TKI treatment in EGFR mutation-enriched patients.

Citation Format: Youngjoo Lee, Kyong-Ah Yoon, Jin Soo Lee. An association between genetic variant in Epidermal Growth Factor (EGF) gene and survival benefit from epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) in never smokers with advanced adenocarcinoma of the lung. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4849. doi:10.1158/1538-7445.AM2013-4849
Abstract 4937: KIF3A promotes prostate cancer cell proliferation, migration, and invasion via activation of Dvl2 and the Wnt signaling pathway.
Authors: Liu Zun , Rebowe Ryan E. , Wang Zemin , Wang Zehua , DePaolo John S. , Guo Jianhui , Li Yingchun , Qian Chiping , Liu Wanguo
Published: August 2014

Aberrant activation of the Wnt signaling pathway is a key event in advanced prostate cancer. However, the mutations in APC complex that activate Wnt signaling in many other cancers are rare in prostate cancer. Other molecular mechanisms that regulate the Wnt signaling pathway in prostate cancer remain to be elucidated. Here we show that KIF3A, a subunit of kinesin-II motor protein, functions as an agonist of the Wnt signaling pathway in prostate cancer. KIF3A is up-regulated in the majority of human prostate cancer cell lines and primary tumor biopsies. The expression levels of KIF3A correlate with higher Gleason score, TNM grade, and metastatic status of prostate cancer. Moreover, overexpression of KIF3A stimulates the benign p69 prostate cell growth, whereas depletion of KIF3A by shRNA suppresses LNCaP cell proliferation, anchorage-independent growth, and cell migration/invasion. Mechanistically, we demonstrate that KIF3A activates DVL2 and induces transcriptional activation of Cyclin D1, HEF1, and MMP9 in prostate cancer cell lines that are required for cancer cell proliferation, migration, and invasion. Taking together, these findings indicate that KIF3A is a potent agonist of Wnt signaling in prostate cancer and suggest that up-regulation of KIF3A is causal of aberrant activation of Wnt signaling in advanced prostate cancer. Inactivation of KIF3A may improve survival of patients with advanced prostate cancer in which Wnt signaling is activated.

Citation Format: Zun Liu, Ryan E. Rebowe, Zemin Wang, Zehua Wang, John S. DePaolo, Jianhui Guo, Yingchun Li, Chiping Qian, Wanguo Liu. KIF3A promotes prostate cancer cell proliferation, migration, and invasion via activation of Dvl2 and the Wnt signaling pathway. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4937. doi:10.1158/1538-7445.AM2013-4937
Abstract 4989: Conditional NFAT activation promotes prostate oncogenesis through cell autonomous and non-cell autonomous effects.
Authors: Manda Kalyan Reddy , Tripathi Piyush , Wang Yinqiu , Humphrey Peter A. , Pan Minggui , Chen Feng
Published: August 2014

Although the activation of nuclear factor of activated T Cells (NFAT) has been linked to cellular transformation and its upregulation has been found in multiple tumor types, its potential role in prostate cancer (PCa) is unknown. We have recently found that NFATc1 activation in prostate epithelium results in prostatic intraepithelial neoplasia (PIN) that progresses to prostate adenocarcinoma in a Doxycycline (Dox)-inducible murine model. The adenocarcinoma primarily consists of Cytokeratin 8 (CK8)+, Androgen receptor (AR)+ luminal epithelial cells and reactive stroma with features similar to the most common types of human PCa. Both cells with NFATc1 activation and neighboring cells without NFATc1 activation have significant upregulation of c-Myc and activation of Stat3. Besides known and suspected NFATc1 targets, such as Spp1, we have revealed the early upregulation of a number of cytokines and cytokine receptors, as key molecular components of an inflammatory and pro-mitogenic microenvironment that promotes both NFATc1+ and NFATc1− cells to participate in tumor formation. Thus, NFATc1 activation has cell autonomous effects on the NFATc1+ cells and non-cell autonomous effects on neighboring cells through secreted factors, resulting in eciprocal prostatic epithelial-stromal interactions for promoting PCa development. Our studies present the first direct in vivo evidence that NFATc1 activation can induce PCa resembling human PCa and provide a versatile platform to further study the pathogenic mechanism of PCa.

Citation Format: Kalyan Reddy Manda, Piyush Tripathi, Yinqiu Wang, Peter A. Humphrey, Minggui Pan, Feng Chen. Conditional NFAT activation promotes prostate oncogenesis through cell autonomous and non-cell autonomous effects. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4989. doi:10.1158/1538-7445.AM2013-4989
Abstract 5060: Discrepancy between MMP-9 and its fibronectin-like domain in tumor growth and invasion.
Authors: Dayer Cynthia
Published: August 2014

Tumors are often compared to wounds that do not heal, where the crosstalk between tumor cells and their surrounding stroma is crucial at all stages of development from the initial primary growth to metastasis. Similar to wound healing, fibroblasts in the tumor stroma differentiate into myofibroblasts, also referred to as “cancer-associated fibroblasts” (CAFs), primarily, but not exclusively, in response to transforming growth factor-β (TGF-β). Myofibroblasts in turn enhance tumor progression by remodeling the stroma. Among molecules implicated in stromal remodeling, matrix metalloproteinases (MMPs), and MMP-9 in particular, play a prominent role. However, the mechanisms that regulate MMP-9 activation and function remain poorly understood. Recent evidence indicates that tumor cell surface association of MMP-9 is an important event in its activation, and more generally in tumor growth and invasion.

Based on these observations, we addressed the potential association of MMP-9 activity with cell-surface recruitment on human fibroblasts. We show for the first time that recruitment of MMP-9 to the MRC5 fibroblast cell surface occurs through the fibronectin-like (FN) domain, found only in MMP-9 and MMP-2. Functional assays suggest that the active form of MMP-9 triggers α-smooth muscle actin (αSMA) expression in resting fibroblasts that reflects myofibroblast differentiation, possibly through TGF-β activation.

Xenograft experiments using HT1080 fibrosarcoma or MDA-MD231 breast adenocarcinoma cells stably expressing the FN domain of MMP-9 in NOD/SCID mice revealed no changes in primary tumor growth. However, in the context of metastasis, expression of the FN domain by these same tumor cells dramatically increases their metastatic proclivity whereas expression of wt MMP-9 either promotes no change or actually reduces the number of metastases. Curiously, xenografts of SW480 colorectal adenocarcinoma cells stably expressing the FN domain of MMP-9 display reduced growth at both the primary (subcutaneous) injection site and the lungs, in experimental metastasis assays, of NOD/SCID mice. This discrepancy is believed to be due to the FN domain that may modulate MMP-9 activity and TGF-β activation in different ways, depending on the tumor cell type.

These observations suggest a dual role of MMP-9 and its FN domain in primary tumor growth and metastasis, underscoring the notion that the effect of MMP-9 on tumor cells may depend on their response to TGF-β activation and highlighting possible protective effects of MMPs in tumor progression and invasion.

Citation Format: Cynthia Dayer. Discrepancy between MMP-9 and its fibronectin-like domain in tumor growth and invasion. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5060. doi:10.1158/1538-7445.AM2013-5060
Abstract 5078: Response-guided anti-angiogenic therapy for hepatocellular carcinoma recurrence with clinically available safety agent.
Authors: Yoshiji Hitoshi , Noguchi Ryuichi , Kaji Kosuke , Aihara Yosuke , Namisaki Tadashi , Moriya Kei , Douhara Akitoshi , Kitade Mitsuteru , Fukui Hiroshi
Published: August 2014

The incidence of hepatocellular carcinoma (HCC) is increasing not only in Asia but also in Europe and United States. Since the high-risk group of either primary or secondary HCC development seems to be clearer than in the other types of tumors, it is likely that chemopreventive agents will be beneficial in improving the prognosis of HCC. No chemopreventive agent, however, has been approved against HCC yet. Since neovascularization plays a pivotal role in HCC, an angiostatic agent is considered as one of the promising approaches. Although an anti-angiogenic agent, sorafenib has approved for advanced heparocellular carcinoma (HCC), most of patients show adverse effects in liver cirrhosis. Insulin resistance (IR) is reportedly involved in the progression of HCC partly mediated with neovascularization. The aim of this study was to develop the response-guided anti-angiogenic therapy with a clinically used safe agent; branched-chain amino acid (BCAA), which reportedly possess anti-angiogenic and IR improving activities, for the cumulative recurrence of HCC after curative therapy. BCAA (Livact; 12 g/day) was administered after the curative therapy for HCC, and 60-month follow-up revealed that the BCAA markedly inhibited the cumulative recurrence of HCC under the condition of IR along with suppression of VEGF. Chronological studies reveled that the VEGF and AFP-L3 were significantly decreased in patients without recurrence at 6 and 12 months after the treatment, respectively. The soluble VEGFR (sVEGFR)-2 was significantly suppressed only in patients without recurrence, whereas sVEGFR-1 was not. A marked suppression of sVEGFR-2 could be achieved within only three months after the treatment. Collectively, since BCAA is widely and safely used in the clinical practice to treat the patients with chronic liver diseases, BCAA may represent a new strategy for secondary chemoprevention for the HCC patients with IR. Moreover, our findings suggest that sVEGFR-2 could become a useful response-guided predictive marker of anti-angiogenic therapy against HCC by BCAA under the condition of IR.

Citation Format: Hitoshi Yoshiji, Ryuichi Noguchi, Kosuke Kaji, Yosuke Aihara, Tadashi Namisaki, Kei Moriya, Akitoshi Douhara, Mitsuteru Kitade, Hiroshi Fukui. Response-guided anti-angiogenic therapy for hepatocellular carcinoma recurrence with clinically available safety agent. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5078. doi:10.1158/1538-7445.AM2013-5078
Abstract 5126: A poor prognosis subtype of HNSCC is consistently observed acrossmethylome, transcriptome and miRNome analysis.
Authors: Jung Alain C. , Job Sylvie , Ledrappier Sonia , Macabre Christine , Abecassis Joseph , de Reynies Aurélien , Wasylyk Bohdan
Published: August 2014

Background: Distant metastasis after treatment is observed in about 20% of Head and Neck Squamous Cell Carcinoma (HNSCC). In the absence of any validated robust biomarker, patients at higher risk for metastasis cannot be provided with appropriate therapy. In order to identify prognostic HNSCC molecular subgroups and potential biomarkers, we have performed genome-wide integrated analysis of four omic sets of data.

Material and methods: Using state-of-the art technologies, a core of 45 metastasizing and 52 non-metastasizing HNSCC patient samples were analyzed at four different levels: gene expression (transcriptome), DNA methylation (methylome), DNA copy number (genome) and miRNA expression (miRNome). Molecular subgroups were identified by a model-based clustering analysis, and their clinical relevance was evaluated by survival analysis.

Results: Transcriptome, methylome and miRNome patient subgroups with shorter metastasis-free survival were identified. A contingency analysis uncovered a R1 group of common tumors, which predicts metastasis occurrence with a higher statistical power than individual omic data sets. R1 and non-R1 samples display similar DNA copy number landscapes, but more frequent chromosomal aberrations are observed in the R1 cluster (especially loss at 13q14.2-3). R1 tumors are characterized by alterations of signaling pathways involved in cell-cell adhesion, EMT, immune response and apoptosis.

Conclusions: Integration of data across several omic profiles leads to better selection of patients at risk, identification of relevant molecular pathways of metastasis, and potential to discover biomarkers and drug targets.

Citation Format: Alain C. Jung, Sylvie Job, Sonia Ledrappier, Christine Macabre, Joseph Abecassis, Aurélien de Reynies, Bohdan Wasylyk. A poor prognosis subtype of HNSCC is consistently observed acrossmethylome, transcriptome and miRNome analysis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5126. doi:10.1158/1538-7445.AM2013-5126
Abstract 5163: MMP3 increases expression of CXCL7.
Authors: Ju Donghong , Kosir Mary A.
Published: August 2014

MMP3 leads to epithelial-mesenchymal transformation (EMT) and malignant transformation of cultured cells. Previously, increased expression of the CXC chemokine CXCL7 was associated with EMT. The hypothesis tested is that MMP3 increases the expression of CXCL7

Methods: Human breast cells (MCF10AT (proliferative), MCFDCIS.com (in situ), MCFCA1a (malignant)) cultured in serum free medium were exposed to MMP-3 for 4 days, and washed 3 times with PBS, then cultured in MMP-3 free condition medium for three more days. Cells were collected every day for RNA extraction, and analyzed for relative CXCL7 gene expression using RT real time PCR. Cells sampled from day 0 were used as control to normalize the results. GADPH was control. Cells culture supernatant also collected every day for CXCL7 protein level expression. Protein concentrations were measured by enzyme-linked immunosorbent assay (ELISA) using human CXCL7/NAP-2 Duo set ELISA Development System.

Measurements were done in triplicate; results are representative of three experiments.

Results: When exposed to MMP-3, the CXCL7 RNA level increased MCF-10AT cells RNA level increased 1.5 fold, and there was no change in CXCL7 protein level in MCF-10AT cells. For MCFDCIS.com cells, the CXCL7 RNA level increased 2.5 fold, and CXCL7 protein level increased 1.6 fold. For MCFCA1a cells, the CXCL7 RNA level increased 2.6 fold, CXCL7 protein level increased 1.5 fold. The peak expression was 3-4 days. Removal of MMP-3 caused CXCL7 expression to return back to baseline in MCF10AT, DCIS.com and malignant Ca.cl1 cells.

Conclusion: The expression of chemokine CXCL7 in breast cell (MCF10AT, DCIS.com, MCFCA1a) was up regulated at both on RNA and protein level (except for MCF10AT) when exposed to MMP-3, and removal of MMP-3 caused CXCL7 expression to return to baseline. The stromal influence by MMP3 on breast cells and breast cancer cells may direct further progression toward the malignant phenotype and provides another therapeutic pathway target.

Citation Format: Donghong Ju, Mary A. Kosir. MMP3 increases expression of CXCL7. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5163. doi:10.1158/1538-7445.AM2013-5163
Abstract 5218: Development of Anti-CD47 therapy for pediatric brain tumors.
Authors: Gholamin Sharareh , Mitra Siddhartha S. , Richard Chase Elliott , Achrol Achal , Kong Doosik , Shin Jun Jae , Monje-Deisseroth Michelle , Cho Yoon-Jae , Weissman Irving , Cheshier Samuel H.
Published: August 2014

Introduction: Extensive molecular characterization of Brain tumors done in recent years has proven valuable for tumor classification, risk stratification and outcome prediction for current treatment modalities. However the current standard of care has not improved the prognosis and carries an increased risk of cognitive impairments for children with brain tumors. A characteristic feature of tumor progression and recurrence is its ability to evade the immune system. Our hypothesis is that by modulating the innate immune system we can enhance the ability of macrophages to ‘eat and kill’ brain cancer cells. Recent data suggests that the interaction between the cell surface antigen CD47 and its binding partner SIRPα is a mechanism by which non-solid and solid tumors can evade the innate immune system. To see if anti-CD47 therapy is a potential therapy in malignant pediatric brain tumors we first looked at CD47 expression in freshly isolated patient and postmortem samples from 4 different tumor types; Diffused Intrinsic Pontine Glioma, Medulloblastoma, Ependymoma and ATRTs. We tested the hypothesis that blocking CD47- SIRPα interaction induced phagocytosis in an in-vitro phagocytosis assay. We next established orthotopic xenografts models in immune compromised mice and treated them with anti-CD47 humanized antibody, which is currently being developed for clinical trials in hematopoietic and non-CNS malignancies.

Result: CD47 expression was upregulated in all tumor types and was present in >90% of the cells in high grade tumors. Increased CD47 expression was observed in CD15+ and CD133+ putative cancer stem cell population. Blocking the CD47- SIRPα interaction increases tumor phagocytosis by macrophages in-vitro. Systemic treatment with anti-CD47 antibody significantly reduced tumor burden in an orthotopic xenograft setting.

Conclusion: Anti-CD47 therapy is a viable and effective treatment modality for pediatric high grade brain tumors.

Citation Format: Sharareh Gholamin, Siddhartha S. Mitra, Chase Elliott Richard, Achal Achrol, Doosik Kong, Jun Jae Shin, Michelle Monje-Deisseroth, Yoon-Jae Cho, Irving Weissman, Samuel H. Cheshier. Development of Anti-CD47 therapy for pediatric brain tumors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5218. doi:10.1158/1538-7445.AM2013-5218
Abstract 5282: The role of T3-regulated microRNAs in human hepatoma cells.
Authors: Huang Ya-Hui , Lin Yang-Hsiang , Chen Ching-Ying , Lin Kwang-Huei
Published: August 2014

Thyroid hormone (T3)/thyroid hormone receptor (TR) signaling regulates cellular growth, development and differentiation, which play important roles in vivo. T3/TR signaling promotes invasion while suppressing proliferation of hepatoma cells. MicroRNAs (miRNA) are small, short, endogenously expressed non-coding RNAs that negatively regulate gene expression. Aberrant miRNA expression has been observed in various tumor types, including hepatocellular carcinoma. At present, it is unclear whether T3/TR signaling regulates miRNAs in hepatoma cells. In our laboratory, miRNA profiling analysis using the high-throughput stem-loop RT-qPCR method revealed upregulation of miR-21 by T3/TR. A native thyroid hormone response element was identified in the primary miR-21 promoter region using promoter and chromatin immunoprecipitation assays. Overexpression of miR-21 promoted hepatoma cell migration and invasion, similar to that observed with T3 stimulation in hepatoma cells. In addition, anti-miR-21-induced suppression of cell migration was rescued by T3. T-cell lymphoma invasion and metastasis 1 (Tiam1), involved in cell migration and invasion, was identified as a miR-21 target, and downregulated by T3. Tiam1 was upregulated upon knockdown of miR-21, and conversely, downregulated upon miR-21 overexpression. Further, knockdown of Tiam1 enhanced migration and invasion may through increase of vimentin and MMP2 expression in hepatoma cells. The correlation among T3/TR, miR-21 and Tiam1 was observed in vivo is similar with in vitro. These findings indicate that miR-21 is directly upregulated by T3. Moreover, T3-mediated enhancement of hepatoma cell migration and invasion may through miR-21-targeted downregulation of Tiam1.

Citation Format: Ya-Hui Huang, Yang-Hsiang Lin, Ching-Ying Chen, Kwang-Huei Lin. The role of T3-regulated microRNAs in human hepatoma cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5282. doi:10.1158/1538-7445.AM2013-5282
Abstract 5289: Identification of basal-like subtype-specific MicroRNAs in lymph node negative breast cancer .
Authors: Shi Wei , Bruce Jeff , Yan Rui , Liu Fei-Fei
Published: August 2014

Background:

Breast cancer is a heterogeneous disease both clinically and biologically. With the recent recognition of the many different molecular subtypes, a desire to more specifically categorize tumors to allow tailoring of treatment to individual patients has developed. One particularly aggressive and treatment-resistant group is the “basal-like” subtype. However, the molecular events driving this subtype of breast cancer are largely unknown. Here, we propose to examine whether MicroRNAs specifically expressed in basal-like breast cancers contribute to their development and progression.

Materials and Methods:

Fifty-four formalin fixed paraffin embedded primary tumour samples from lymph-node negative breast cancer patients who were participants in a phase III randomized trial of tamoxifen (Tam) +/- whole breast radiation were collected. Intrinsic molecular subtyping was determined using semi-quantitative analysis of ER, PR, Ki-67, HER2, EGFR and cytokeratin (CK) 5/6 on tissue microarrays (TMAs) constructed from the tumor blocks. Patients were classified into the following categories: luminal A, luminal B, luminal-HER2, HER2 enriched or basal-like phenotypes. The median follow-up for this cohort was 10 years. The expression of 365 miRNAs was measured in these samples using the qRT-PCR based Taqman low density array human miRNA panel (Applied Biosystems). Data were validated in cell line models and further analyzed using level 3 TCGA breast cancer data.

Results:

Twenty-three miRNAs were identified at a significantly different expressed level in basal-like breast cancers (n = 6) compared with other subtypes (n = 48). This differential expression was validated in publically available RNA-Seq data generated by the TCGA for 20 of the 23 miRNAs. Eight of these twenty miRNAs were found to correlate with corresponding copy-number alterations at their genomic loci using the data generated by TCGA. Further analysis is currently underway to determine the potential contribution of methylation to the expression differences observed. The putative target genes and pathways involved will also be analyzed.

Conclusions:

MiRNAs are known to play numerous roles in cancer development and progression. In the current study we report several miRNAs with distinct expression patterns in basal-like breast cancers. Further characterization of the phenotypic impact of this deregulation may lead to a better understanding of this aggressive subtype and provide valuable insight needed to develop novel targeted therapies to treat these tumours.

Citation Format: Wei Shi, Jeff Bruce, Rui Yan, Fei-Fei Liu. Identification of basal-like subtype-specific MicroRNAs in lymph node negative breast cancer . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5289. doi:10.1158/1538-7445.AM2013-5289
Abstract 5299: Evaluation of apolipoprotein E polymorphism on the aggressive potential of prostate cancer cell lines.
Authors: Ifere Godwin O. , Demark-Wahnefried Wendy , Desmond Renee , Nagy Tim R.
Published: August 2014

Introduction: Defective cholesterol-modulating apolipoprotein E (ApoE) isoforms could exacerbate prostatic neoplasia. Recent evidence strongly suggests that specific genetic variants of ApoE control peripheral tissue cholesterol efflux. Defects in ApoE may result in reduced prostate cancer cell cholesterol efflux and enriched concentrations of cholesterol in membranes by reducing clearance. Membrane-rich cholesterol could potentially activate cancer-driven signaling processes.

Objective: To determine the relevance of specific ApoE phenotypes as risk factors of aggressive prostate cancer based on their influence on reverse cholesterol transport.

Methods: We evaluated ApoE genotypes of the weakly tumorigenic or nonaggressive prostate cancer cell lines (LNCaP and MDA PCA 2b), and the moderate and highly tumorigenic or aggressive prostate cancer cell lines (PC-3 and DU-145). We examined whether ApoE2/E4 variants were associated with tumor aggressiveness via the expression of caveolin-1(cav-1), a cholesterol-regulated pro-carcinogenic gene.

Results: We observed that the nonaggressive prostate cancer cell lines carried either the Apo ε3/ε3 or ε3/ε4 alleles, while the aggressive prostate cancer cell lines carried the Apo ε2/ε4 alleles. Our results showed a two-fold increase in the expression of cav-1 in the aggressive cell lines, as opposed to the nonaggressive ones. There was a significantly (p<0.01) greater cholesterol efflux from the nonaggressive cells lines compared to the aggressive cells. Also, the aggressive cell lines retained and localized more cholesterol in their membranes as opposed to the less aggressive cell lines.

Conclusions: These results suggest an association between cholesterol status and the aggressive potential of prostate cancer cells. It also highlights the need for further investigation into whether ApoE2/E4 is the dysfunctional phenotype, and whether it has the potential as a biomarker of aggressive prostate cancer.

GRANT SUPPORT: NIH R25CA047888

Citation Format: Godwin O. Ifere, Wendy Demark-Wahnefried, Renee Desmond, Tim R. Nagy. Evaluation of apolipoprotein E polymorphism on the aggressive potential of prostate cancer cell lines. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5299. doi:10.1158/1538-7445.AM2013-5299
Abstract 5316: Significant changes in D2-like dopamine gene receptors expression associated with non- small -cell lung cancer: A case control study.
Authors: Sheikhpour Mojhgan , Ahangari Ghasem , Sadeghizadeh Majid , khodadad Kian
Published: August 2014

Introduction : Lung cancer is the leading cause of cancer-related death and a major health problem worldwide. Approximately 75% of lung tumors are non - small - cell lung cancer (NSCLC) .The alterations of five different Dopamine receptor genes expression (DRD1,DRD2,DRD3,DRD4 and DRD5) in certain diseases of immune and nervous systems have been reported and play multiple role in various disorders such as cancer. Previous controlled studies demonstrated a relationship between changes in dopamine receptor gene expression profiles and clinical improvement. In the present study, it was hypothesized that the dopamine receptor gene expression profile of peripheral blood mononuclear cells in NSCLC patients could be different from those normal individuals which probably increases the risk of NSCLC.

Material and methods: We investigated dopamine receptors gene expression changes in peripheral Blood Mononuclear cells (PBMC) of 30 NSCLC patients compared to normal individuals . The PBMC was separated from normal whole blood by Ficoll-hypaque and then the total cellular RNA was extracted and cDNA was synthesized, Real Time PCR was done By using primer pairs specific for five dopamine receptors mRNA and β-actin as internal control . Finally the specificities of the obtaine PCR products for the respective dopamine receptors fragments were comfirmed by sequencing and expression comparing of two group (patients and normal cases) was done by SPSS analysis .Results: the results show that DRD2, DRD3 and DRD4 expression changes profile were significant change in NSCLC patients compared to normal individuals . but DRD1,DRD5 genes didn't show any significant and changes . Conclusions: We conclude that there is a quantitative significant difference of D2-like dopamine receptor genes expression in non small cell lung cancer . On the other hand these significant differences and profiles could contribute in diagnosis, treatment and survey and monitoring of non small cell lung cancer .In addition it could be a lead for design of future therapeutic strategies

Key words: Dopamine receptors - Non-small cell lung cancer - Gene expression

Citation Format: Mojhgan Sheikhpour, Ghasem Ahangari, Majid Sadeghizadeh, Kian khodadad. Significant changes in D2-like dopamine gene receptors expression associated with non- small -cell lung cancer: A case control study. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5316. doi:10.1158/1538-7445.AM2013-5316
Abstract 5356: EZH2-mediated H3K27me3 is involved in epigenetic repression of DLC1 (Deleted in liver cancer 1) in human cancers.
Authors: Au Leung Kuen , Wong Carmen Chak-Lui , Lee Joyce Man-Fong , Wong Chun-Ming , Ng Irene Oi-Lin
Published: August 2014

Enhancer of zeste homolog 2 (EZH2), the histone methyltransferase of the Polycomb Repressive Complex 2 catalyzing histone H3 lysine 27 tri-methylation (H3K27me3), is frequently up-regulated in human cancers, including hepatocellular carcinoma (HCC). In this study, we identified the tumor suppressor Deleted in liver cancer 1 (DLC1) as a target of repression by EZH2-mediated H3K27me3. DLC1 is a GTPase-activating protein for Rho family proteins. Inactivation of DLC1 results in hyper-activated Rho/ROCK signaling and is implicated in actin cytoskeleton reorganization to promote cancer metastasis. By chromatin immunoprecipitation assay, we demonstrated that H3K27me3 was significantly enriched at the DLC1 promoter region of a DLC1-nonexpressing HCC cell line, MHCC97L. Depletion of EZH2 in MHCC97L by short hairpin RNA reduced H3K27me3 level at DLC1 promoter and induced DLC1 gene re-expression. Conversely, transient overexpression of GFP-EZH2 in DLC1-expressing Huh7 cells reduced DLC1 mRNA level with a concomitant enrichment of EZH2 on DLC1 promoter. An inverse relation in the expression pattern between EZH2 and DLC1 was observed in the liver, lung, breast, prostate, and ovarian cancer tissues. Treating cancer cells with the EZH2 small molecular inhibitor, 3-Deazaneplanocin A (DZNep), restored DLC1 expression in different cancer cell lines, indicating that EZH2-mediated H3K27me3 epigenetic regulation of DLC1 was a common mechanism in human cancers. Importantly, we found that DZNep treatment inhibited HCC cell migration through disrupting actin cytoskeleton network, suggesting the therapeutic potential of DZNep in targeting cancer metastasis. Taken together, our study has shed mechanistic insight into EZH2-H3K27me3 epigenetic repression of DLC1 and advocated the significant pro-metastatic role of EZH2 via repressing tumor and metastasis suppressors.

Citation Format: Leung Kuen Au, Carmen Chak-Lui Wong, Joyce Man-Fong Lee, Chun-Ming Wong, Irene Oi-Lin Ng. EZH2-mediated H3K27me3 is involved in epigenetic repression of DLC1 (Deleted in liver cancer 1) in human cancers. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5356. doi:10.1158/1538-7445.AM2013-5356
Abstract 5371: Epigenetic regulation in skin cancer: global DNA methylation profiling in DMBA/TPA induced mice.
Authors: Lee Jong Hun , Shu Limin , Huang Mou-Tuan , Conney Allan H. , Kong Ah-Ng Tony
Published: August 2014

Hypermethylation of CpG islands in the promoter regions of tumor suppressor genes is postulated to be the most relevant epigenetic changes resulting in gene silencing of tumor suppressor gene in cancer. Transcriptionally repressed genes by epigenetic regulation can be reactivated by epigenetic modifications because these silenced genes are still genetically intact. Thus, it is relevant to profile global DNA methylation changes that occur in early stages of carcinogenesis (Jones & Baylin, 2002). To begin addressing this issue, we conducted global profiling of DNA methylation changes in a well-established multistage skin carcinogenesis model using methylated DNA immunoprecipitation (Medip) coupled with CpG island microarray. Tumorigenesis was induced by sequential topical application of mutagen 7,12-Dimethylbenz(a)anthracene (DMBA), and promoting agent 12-O-tetra-decanoylphorbol-13-acetate (TPA), on the dorsal skin of normal CD-1 mice. After 12 weeks, animals were sacrificed and DNA extracted from papillomas. Methylation enriched DNA fragments were pulled down and hybridized to CpG microarray covering 16,030 CpG islands (Agilent, Santa Clara, CA) The results show that significant changes in DNA methylation profile between normal skin and DMBA/TPA induced papilloma, analyzed by Genespring and Bioconductor. After normalization of probe with the gene, 186 genes from 9,867 of up-regulated methylation genes showed at least average 2-fold changes and these include some DNA damage and repair, oxidative stress and inflammatory genes such as Gadd45b and Arnt2. In summary, DMBA/TPA induced mouse skin papilloma undergo extensive aberrant CpG island hypermethylation. Genes detected from Medip-chip assay may be utilized as a potential epigenetic tool to study skin cancer.

Jones, P. A., & Baylin, S. B. (2002). The fundamental role of epigenetic events in cancer. Nature Reviews Genetics, 3, 415-428.

Citation Format: Jong Hun Lee, Limin Shu, Mou-Tuan Huang, Allan H. Conney, Ah-Ng Tony Kong. Epigenetic regulation in skin cancer: global DNA methylation profiling in DMBA/TPA induced mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5371. doi:10.1158/1538-7445.AM2013-5371
Abstract 5429: Robust in vitro cancer metabolism in non-tumorigenic microglial cells.
Authors: Flores Roberto , Kami Kenjiro , Seyfried Thomas
Published: August 2014

It is well documented that energy metabolism differs between cancerous tissue and normal tissue for a large variety of human and animal tumors. The murine J2 retrovirus contributes to oncogenesis in macrophage cells. The BV-2 microglia cells were immortalized using the v-raf/v-myc carrying J2 retrovirus. These cells, however, do not form tumors when grown in vivo in the natural C57BL/6 host. Consistent with v-raf/v-myc expression, the in vitro metabolic and proliferation rate of the BV-2 cells greatly exceeded that of the tumorigenic and metastatic VM-M3 microglia cells when grown in DMEM containing 25 mM glucose. When controlling for proliferation and ATP production, the BV-2 cells produced more lactate than did the VM-M3 cells under aerobic conditions, thus demonstrating a robust Warburg effect in the BV-2 cells. The Warburg effect is associated with HIF stabilization under aerobic conditions. Mutations in the IDH1 gene produce 2-hydroxyglutarate (2HG), a recognized “onco-metabolite” that also contributes to HIF stabilization under aerobic conditions. Mass spectrometry analysis showed that the non-tumorigenic BV-2 cells made 2-HG only under anoxic conditions. 2-HG was not detected under anoxia or normoxia in VM-M3LM cells, a less malignant clonal variant of VM-M3. It appears that the BV-2 cells have a more robust cancer metabolism than either the high malignant VM-M3 cells or the low malignant VM-M3LM cells. The VM-M3LM cells consumed more oxygen and produced less lactate per ATP than did either the BV-2 cells or the VM-M3 cells, thus displaying a reduced Warburg effect. In other words, the Warburg effect was greatest in the non-tumorigenic BV-2 cells, was lowest in the low malignant VM-M3LM cells, but was intermediate in the high malignant VM-M3 cells. The results suggest that over expression of the v-raf/v-myc oncogenes, production of the IDH1 onco-metabolite, and a robust Warburg effect are not sufficient to drive tumorigenesis in the BV-2 cells. Supported in part from Boston College Res. Expense Fund.

Citation Format: Roberto Flores, Kenjiro Kami, Thomas Seyfried. Robust in vitro cancer metabolism in non-tumorigenic microglial cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5429. doi:10.1158/1538-7445.AM2013-5429

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 5462: MEDI3185, a potent anti-CXCR4 antibody, inhibits tumor cell migration, signaling and tumor growth in preclinical models.
Authors: Kamal Adeela , Wang Youzhen , Steiner Philipp , Mazzola Anne-Marie , Wetzel Leslie , Passino Melissa , McDermott Brenda , Huang Keven , Bedian Vahe , Greenberg Norman
Published: August 2014

The chemokine receptor CXCR4 is a seven-transmembrane G-protein coupled receptor that mediates chemotaxis and cell migration upon stimulation via its ligand, stromal-derived factor 1 (SDF-1), also called CXCL12. CXCR4 is normally expressed on bone marrow stem and progenitor cells, various circulating lymphocytes, endothelial precursor cells, tissue macrophages and fibroblasts but the aberrant overexpression of CXCR4 is linked to various hematological malignancies, solid tumors and metastatic neoplasms. Moreover, CXCR4 overexpression is correlated with poor prognosis in many types of cancer, including breast, ovarian, colon, pancreatic, AML and glioblastomas. CXCR4 inhibition using siRNA, small-molecule and peptide inhibitors has demonstrated that it can inhibit tumor growth by blocking tumor cell survival/proliferation, metastasis, angiogenesis and tumor immune infiltrates. Here we describe a novel, fully human, antagonistic antibody to CXCR4, MEDI3185, which blocks SDF-1 binding to CXCR4. MEDI3185 has picomolar binding affinity to human CXCR4 and exhibits no significant binding to other chemokine receptors such as CCR4 or CXCR3. In vitro studies demonstrated that MEDI3185 inhibited tumor cell migration, blocked SDF-1 induced tumor cell signaling and induced apoptosis of tumor cells. In preclinical human tumor xenograft models in mouse, MEDI3185 showed single-agent tumor growth inhibition in multiple myeloma and B-cell Burkitt's lymphoma models and had combination activity in ovarian models. In addition, MEDI3185 extended survival as combination therapy in mouse models of CLL and also blocked lung tumor burden in a disseminated ovarian model. Combined, these data suggest that MEDI3185 is a potent CXCR4 antibody for the treatment of both hematological and solid tumors because it has pleiotropic effects on tumor biology that may enhance the efficacy of the current standard of care.

Citation Format: Adeela Kamal, Youzhen Wang, Philipp Steiner, Anne-Marie Mazzola, Leslie Wetzel, Melissa Passino, Brenda McDermott, Keven Huang, Vahe Bedian, Norman Greenberg. MEDI3185, a potent anti-CXCR4 antibody, inhibits tumor cell migration, signaling and tumor growth in preclinical models. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5462. doi:10.1158/1538-7445.AM2013-5462