bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4616: Chemo-sensitisation in epithelial ovarian cancer cell lines by targeting Ankyrin Repeat Domain 1 (ANKRD1)
Authors: Lei Ying , Henderson Beric Ross , Emmanuel Catherine , Harnett Paul , Fazio Anna de
Published: May 2015

Primary and acquired chemotherapy resistance remains a major challenge in the treatment of epithelial ovarian cancer and there are currently few targeted treatments that improve clinical outcome. We screened for genes that could potentially be targeted to sensitise, or re-sensitise, ovarian cancer cells to chemotherapy and identified a novel candidate, Ankyrin Repeat Domain 1 (ANKRD1), not previously described in ovarian cancer. ANKRD1 is a putative transcription factor with roles in muscle development, inflammation and response to cell stress. ANKRD1 is a target of TGF-β/Wnt signalling, and is also a binding partner of Titin (TTN) which been identified as a candidate with a high probability of carrying ‘driver’ mutations in ovarian cancer. ANKRD1 was expressed in serous ovarian cancer samples and higher tumor levels were found in patients with worse outcome (overall survival, p=0.013), (Scurr et al, Clin Cancer Res 2008; 14: 6924-32). In this study we showed that ANKRD1 levels decrease in response to cisplatin, in concert with induction of PARP cleavage and apoptosis in human ovarian cancer cell lines, COLO316, PEO14 and OAW42. We showed that decreasing ANKRD1 expression using siRNA (ON-TARGETplus Human ANKRD1 siRNA) increased cisplatin-induced apoptosis. Increased cisplatin sensitivity was associated with endoplasmic reticulum (ER) stress, evidenced by induction of GRP78, GADD153 and increased intracellular Ca2+ release. We have screened drug libraries (Prestwick library) for agents that sensitise ovarian cancer cell lines to platins, via an ANKRD1 related mechanism, and have identified several potential candidate molecules. Taken together, these results suggest that ANKRD1 plays a significant role in the regulation of apoptosis in human ovarian cancer cells, and is a potential molecular target to enhance sensitivity of ovarian cancer to chemotherapy.

Citation Format: Ying Lei, Beric Ross Henderson, Catherine Emmanuel, Paul Harnett, Anna de Fazio. Chemo-sensitisation in epithelial ovarian cancer cell lines by targeting Ankyrin Repeat Domain 1 (ANKRD1). [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4616. doi:10.1158/1538-7445.AM2014-4616
Abstract 4638: NVP-CGM097: a novel p53-Mdm2 inhibitor exhibiting potent antitumor activity in mouse models of human cancer
Authors: Ferretti Stéphane , Berger Marjorie , Rebmann Ramona , Santacroce Francesca , Sterker Dario , Jensen Michael , Masuya Keiichi , Jeay Sébastien
Published: May 2015

Although functional inactivation of the tumor suppressor p53 is a frequent genetic event, half of all human tumors express wild-type p53. Stabilization of p53 by antagonizing its negative regulator mouse double minute 2 (Mdm2) leads to selective induction of the p53 pathway, thus offering a promising therapeutic opportunity.

NVP-CGM097 binds in the p53 pocket on the surface of Mdm2 and effectively disrupts the interaction between p53 and Mdm2. The aim of this study was to determine the pharmacokinetic (PK), pharmacodynamic (PD) and efficacy profile of this novel p53-Mdm2 inhibitor in mouse models of human cancer. During the medicinal chemistry discovery efforts, the compound series was optimized for oral administration. The selected compound, NVP-CGM097, was well absorbed in mice where the maximum plasma concentration (Tmax) was observed about 3 hours post dose followed by a prolonged absorption phase. PD effects were determined by measuring the expression levels of p53 target genes including p21, PUMA and Mdm2 in tumors. The expression levels of p53 target genes correlated with the concentrations of NVP-CGM097 demonstrating a good PK/PD relationship.

Optimal treatment schedules were investigated in the Mdm2-amplified, p53 wild-type osteosarcoma xenograft model SJSA-1. Interestingly, various schedules were highly efficacious at well-tolerated dose levels. Tumor regression was observed when NVP-CGM097 was administered in schedules ranging from daily to twice per week with no adverse effects. Following dose optimization studies, potent anti-tumor efficacy was observed in a range of p53 wild-type expressing xenograft models including Mdm2 non-amplified orthotopic AML as well as well-differentiated (WDLPS) and dedifferentiated (DDLPS) liposarcomas.

Overall, we demonstrate that the novel p53-Mdm2 inhibitor NVP-CGM097 has a good PK profile and affects PD markers in human tumors leading to tumor regression at well-tolerated dose levels. These data strongly support clinical development of NVP-CGM097.

Citation Format: Stéphane Ferretti, Marjorie Berger, Ramona Rebmann, Francesca Santacroce, Dario Sterker, Michael Jensen, Keiichi Masuya, Sébastien Jeay. NVP-CGM097: a novel p53-Mdm2 inhibitor exhibiting potent antitumor activity in mouse models of human cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4638. doi:10.1158/1538-7445.AM2014-4638
Abstract 4661: The regulatory mechanism of fibronectin in EGFR and/or HER2 positive breast cancer
Authors: Jeon Myeongjin
Published: May 2015

Fibronectin (FN) is a glycoprotein of the extracellular matrix and plays a major role in cell adhesion, migration and oncogenic transformation. Aberrant FN expression is associated with poor prognosis in a variety of cancers including breast cancer. However, the regulatory mechanism between FN and HER2 is not fully elucidated in breast cancer.

Our clinical data showed that both disease free survival (DFS) and overall survival (OS) was lower in EGFR (+) and HER2 (+) breast cancer patients than in EGFR (+) breast cancer patients. To verify the causing of poor prognosis, we analyzed secreted proteins in EGFR and/or HER2 overexpressed MDA-MB231 cells. Our results showed that the level of FN expression was significantly increased in HER2 overexpressed MDA-MB231 cells. On the other hand, the induction of FN by HER2 overexpression was decreased by trastuzumab (HER2 monoclonal antibody) and AG825 (HER2 inhibitor), respectively. In addition, we observed that the expression of FN was dose-dependently increased by EGFR ligands in SKBR3 cells which are expressed endogenous EGFR and HER2. EGF ligands-induced FN expression also decreased through the suppression of HER2 in SKBR3 cell. However, these effects did not detect in trastuzumab-resistant breast cancer cell line, JIMT-1. Taken together, we demonstrated that the induction of FN by HER2 correlates poor clinical outcome in breast cancer patients.

Citation Format: Myeongjin Jeon. The regulatory mechanism of fibronectin in EGFR and/or HER2 positive breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4661. doi:10.1158/1538-7445.AM2014-4661
Abstract 4662: Prognostic value of total lesion glycolysis from 18F-fluorodeoxyglucose positron emission tomography patients with gastric neuroendocrine carcinoma
Authors: Jung Minkyu , Kang Beodeul , Park Ji Soo , Lim Sun Min , Kim Hyo Song , Rha Sun Young , Ahn Joong Bae , Chung Hyun Cheol , Yun Mijin , Cho Arthur
Published: May 2015

Background: Gastric neuroendocrine carcinomas (NET) are very rare, aggressive tumors of the stomach. The objective of this study was to examine the predictive role of pretreatment fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG PET)-assessed metabolic parameter of the whole-body and primary tumors in patients with gastric NET

Method: We conducted a retrospective review of the 25 patients with histopathologically confirmed poorly differentiated NETs of the stomach at our hospital between January 2005 and December 2012. All of the patients underwent 18F-FDG-PET examination at diagnosis. Metabolic parameters [SUVmax, SUVmean, and total lesion glycolysis (TLG)] on pretreatment 18F-FDG-PET were analyzed for survival.

Results: Median follow up duration was 39.4 months (95% CI, 20.0-58.1 months) and median overall survival (OS) was 25.7 months (95% CI, 14.1-37.2 months). When subjects were divided into two groups according to TLG with a cutoff value of 500, the high TLG group showed significantly shorter OS compared to the low TLG group (p=0.006). However, SUVmean and SUVmax had no significant association OS (p = 0.258, and p = 0.52, respectively).

Conclusions: Our findings suggest that TLG may be more useful than SUVmean and SUVmax for predicting OS in patient with NET. The TLG measurement on 18F-FDG-PET imaging could be routinely recommended to advanced gastric cancer with neuroendocrine carcinoma

Citation Format: Minkyu Jung, Beodeul Kang, Ji Soo Park, Sun Min Lim, Hyo Song Kim, Sun Young Rha, Joong Bae Ahn, Hyun Cheol Chung, Mijin Yun, Arthur Cho. Prognostic value of total lesion glycolysis from 18F-fluorodeoxyglucose positron emission tomography patients with gastric neuroendocrine carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4662. doi:10.1158/1538-7445.AM2014-4662
Abstract 4670: Characterization of Met signaling in urothelial cancer of the bladder
Authors: Lee Young H. , Apolo Andrea , Agarwal Piyush K. , Bottaro Donald P.
Published: May 2015

Background: Mounting evidence supports Met as a target in urothelial cancer (UC). Activated Met can promote angiogenesis and tumor growth by upregulating VEGF and may play a role in UC pathogenesis. Cabozantinib inhibits both VEGFR2 and Met pathways. In this study, we assessed cabozantinib's effects on HGF-driven UC cell growth and invasion.

Methods: sMet levels in serum and urine samples from 31 patients with UC (23 metastatic, 8 muscle-invasive) were correlated with stage, presence of visceral metastases and urinary source. The effects of cabozantinib on 6 human UC-derived cell lines were studied in vitro. Intact RT4, TCC-SUP, T24M2, T24M3, J82 and SW780 cells at 80% confluence were serum deprived 16 h, then left untreated or treated with hepatocyte growth factor (HGF) and/or cabozantinib prior to analysis of Met, phospho- (p)Met, pAkt, Akt, pErk and ErK by immunoassay or immunoblotting. Cabozantinib effects on basal and HGF-induced UC cell invasion, proliferation and soft agar growth were measured.

Results: Median serum Met levels were modestly higher in patients with metastatic versus muscle-invasive disease. Urinary Met levels were clearly higher in patients with visceral metastasis (p=0.0111) and in urine from ileal conduits and neobladders compared to normally voided urine, regardless of stage (p=0.0489). Met content in UC cell lines was low in RT4 and higher in T24M2, T24M3, TCC-SUP, J82 and SW780. Basal pMet content was universally low, increased significantly by HGF and this was reversed by cabozantinib. HGF-driven increases in pAkt/Akt and pErK/ErK in all 6 cell lines were reversed by cabozantinib, as were HGF-enhanced UC cell invasion, proliferation and anchorage independent growth.

Conclusions: Median urinary sMet is significantly higher in patients with visceral metastasis and in specimens from ileal conduits and neobladders relative to normally voided urine. UC cell Met content in culture increased with disease grade; HGF stimulated activation of Met and known effectors, and enhanced invasion, growth rate and anchorage-independent grow¬¬¬th; cabozantinib effectively reversed these HGF-driven effects.

Citation Format: Young H. Lee, Andrea Apolo, Piyush K. Agarwal, Donald P. Bottaro. Characterization of Met signaling in urothelial cancer of the bladder. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4670. doi:10.1158/1538-7445.AM2014-4670
Abstract 4674: Finding cancer biomarkers in platelets
Authors: Tjon-Kon-Fat Lee-Ann , Lundholm Marie , Wurdinger Thomas , Thellenberg Camilla , Widmark Anders , Wikström Pernilla , Nilsson Jonas
Published: May 2015

Introduction: In the development of personalized medicine we need a window in to the tumor; it should be minimal invasive, being able to find the disease and follow how the tumor alters its behaviour to circumvent the treatment. Platelets could be one such window, with its ability to sequester tumor-derived material during its passage through the circulation. Material & Methods: With conventional quantitative PCR and microarray techniques “potential” biomarkers were analysed from WTA (whole transcriptome amplified) platelet-derived RNA isolated from cancer patients and healthy controls. Results: We demonstrate that platelets contain several cancer-associated RNA biomarkers. In addition, there was a distinct biomarker signature in platelets from cancer patients of different grades as compared to normal control subjects. Conclusion: These findings are preliminary, and need to be validated in a larger patient cohort. However, they support our earlier findings in gliomas that platelets may carry the needle from the haystack that will give us a window into the cancer biology of individual patients.

Citation Format: Lee-Ann Tjon-Kon-Fat, Marie Lundholm, Thomas Wurdinger, Camilla Thellenberg, Anders Widmark, Pernilla Wikström, Jonas Nilsson. Finding cancer biomarkers in platelets. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4674. doi:10.1158/1538-7445.AM2014-4674
Abstract 4677: Niclosamide analogs for treatment of ovarian cancer
Authors: Haygood Christen L. Walters , Arend Rebecca C. , Li Pui-Kai , Li Yonghe , Kurpad Chandrika , Gangrade Abhishek , Straughn John M. , Buchsbaum Donald
Published: May 2015

Objective: Niclosamide has shown promising activity against ovarian cancer in vitro;, however, it has low bioavailability in vivo. Thus, we investigated the cytotoxicity of niclosamide analogs in combination with carboplatin in ovarian cancer cell lines and against ovarian cancer patient ascites cells.

Methods: Tumorspheres were isolated from ascites collected from 8 patients undergoing ovarian cancer surgery at UAB. Tumorspheres were plated at 10,000 cells per 50 µL into low attachment plates. Eight patient samples were treated concurrently with niclosamide analog 11 (0.1 µM - 5 µM) and carboplatin (5 - 150 μM), while four 4 patient samples were treated with analog 32 (0.1 µM - 5 µM) and carboplatin (5 - 100 μM). After 48 hours, cells were analyzed for viability using the ATPlite assay. Analogs 11 and 32 were also assayed in combination with carboplatin against ovarian cancer cell lines, A2780, A2780.cp20, SKOV3ip2, and SKOV3ip2.TR.

Results: IC50 values for analog 11 ranged from 0.5 - 1 μM in the 8 patient ascites samples. With 150 μM carboplatin alone, patient samples had 40-80% cytotoxicity. In combination with analog 11 at 1 μM, 100% of cells were killed with 100 μM carboplatin in the 8 patient ascites samples. In the established ovarian cancer cell lines, cytotoxicity approached 100% at a dose of 3 - 5 μM of analog 11. In the 4 patient samples treated with analog 32, IC50 values ranged from 0.4 μM - 3 μM. With 100 μM carboplatin alone, patient samples had 40-80% cytotoxicity. In combination with 100 μM carboplatin, nearly 100% of cells were killed at a dose of 1 μM of analog 32. In the cell lines, IC50 values for analog 32 ranged from 0.9 - 2.5 μM. With combination treatment, there was increased cytotoxicity in the chemoresistant cells, A2780.cp20 and SKOV3ip2, compared to carboplatin alone.

Conclusion: These niclosamide analogs produced cytotoxicity both alone and in combination with carboplatin against ovarian cancer cells and tumorspheres from patient ascites. Future studies will evaluate the PK/PD of the analogs.

Citation Format: Christen L. Walters Haygood, Rebecca C. Arend, Pui-Kai Li, Yonghe Li, Chandrika Kurpad, Abhishek Gangrade, John M. Straughn, Donald Buchsbaum. Niclosamide analogs for treatment of ovarian cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4677. doi:10.1158/1538-7445.AM2014-4677
Abstract 4679: Novel pathway mutations in malignant mesothelioma revealed by high-throughput DNA and RNA sequencing
Authors: Miyanaga Akihiko , Masuda Mari , Tsuta Koji , Nakamura Yuka , Asamura Hisao , Gemma Akihiko , Yamada Tesshi
Published: May 2015

Background: Malignant mesothelioma (MM) is an aggressive neoplasm associated with asbestos exposure. Because MM is usually diagnosed at an advanced stage and is largely unresponsive to conventional therapy, it is necessary to identify the signal pathways driving MM and to develop therapeutics specifically targeting these pathways.

Materials and Methods: We carried out RNA sequencing of 12 MM cell lines and 4 clinical samples and exome sequencing of 6 MM cells using a so-called next-generation sequencer. We further performed pathway-targeted genome sequencing of 23 paired samples of MM tumor and normal tissue.

Results: We identified 15 novel fusion genes by RNA sequencing and confirmed them by conventional Sanger sequencing and FISH analyses. We showed that the novel gene was included in both LOH and a fusion gene, and the gene which was critical tumor-suppressor roles caused two hit and was down-regulated. The gene has been identified as a central player of the emerging signaling pathway. We further identified the accumulation of mutations in 35% of the MMs by pathway-targeted genome sequencing.

Conclusions: We indicated the need for the discovering molecular targets in the novel pathway in MMs. Our findings provide novel insights into the pathogenesis of MM and unequivocally demonstrate the power of sequencing technologies in cancer research.

Citation Format: Akihiko Miyanaga, Mari Masuda, Koji Tsuta, Yuka Nakamura, Hisao Asamura, Akihiko Gemma, Tesshi Yamada. Novel pathway mutations in malignant mesothelioma revealed by high-throughput DNA and RNA sequencing. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4679. doi:10.1158/1538-7445.AM2014-4679
Abstract 47: Y-box binding protein 1 is correlated with lymph node metastasis in intestinal type of gastric cancer
Authors: Guo TianTian , Yu Yingnan , Yip George Wai-Cheong , Baeg Gyeong Hun , Thike Aye Aye , Tan Puay-Hoon , Matsumoto Ken , Bay Boon Huat
Published: May 2015

Gastric cancer still ranks among the top ten most common cancer in the world, despite a declining trend in incidence. The multifunctional protein Y Box Binding Protein-1 (YB-1) is known to regulate transcription, translation, DNA repair and cellular response to drug treatment. This study evaluated the correlation between YB-1 protein expression levels with clinico-pathological characteristics in the intestinal type of gastric cancer, which was further validated with in vitro experimentation. Tissue microarrays (TMAs) were constructed from specimens obtained from 167 patients diagnosed with intestinal gastric cancer. Immunohistochemical analysis of the YB-1 protein was performed on the TMAs . The intensity of YB-1 staining in the intestinal type of gastric cancer was scored as absent to strong staining and the percentage of YB-1 immunostained cells was determined. YB-1 immunoreactivity was based on the weighted average intensity (WAI) for each specimen, which was calculated as the sum of percentages of stained cells for each intensity divided by the total percentage of stained cells. siRNA-mediated silencing of the YB-1 gene in MKN7 intestinal gastric cancer cells was verified by RT-PCR. Cell migration and invasion assays were carried out using commercially available kits. YB-1 immunostaining was present in all the cancer tissue samples examined. The YB-1 protein was observed to be predominantly expressed in the cytoplasm of the gastric cancer cells. Univariate analysis revealed that YB-1 immunohistochemical expression was significantly correlated with lymph node status and perforation. However, multivariate analysis showed that only YB-1 protein expression is an independent predictor of lymph node spread. Down-regulation of the YB-1 gene expression in MKN7 cells significantly inhibited cell migration and invasion, processes which are important in the metastatic cascade. The findings suggest that YB-1 expression is associated with metastasis in the intestinal type of gastric cancer and is a potential prognostic biomarker and therapeutic target in this subtype of gastric cancer.

Citation Format: TianTian Guo, Yingnan Yu, George Wai-Cheong Yip, Gyeong Hun Baeg, Aye Aye Thike, Puay-Hoon Tan, Ken Matsumoto, Boon Huat Bay. Y-box binding protein 1 is correlated with lymph node metastasis in intestinal type of gastric cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 47. doi:10.1158/1538-7445.AM2014-47
Abstract 4717: Detection of the Cyclin J (CCNJ) as a new cancer-related gene in human hepatocellular carcinoma by using a method of triple combination array analysis
Authors: Takano Nao , Nomoto Shuji , Hishida Mitsuhiro , Inokawa Yoshikuni , Hayashi Masamichi , Kanda Mitsuro , Okamura Yukiyasu , Nishikawa Yoko , Tanaka Chie , Kobayashi Daisuke , Yamada Suguru , Nakayama Goro , Fujii Tsutomu , Sugimoto Hiroyuki , Koike Masahiko , Fujii Michitaka , Takeda Shin , Kodera Yasuhiro
Published: May 2015

Background: Hepatocellular carcinoma (HCC) has a high rate of recurrence and a poor prognosis. To detect genes correlated with HCC, we have developed a new technique, triple combination array analysis, consisting of methylation array, gene expression array and single nucleotide polymorphism (SNP) array analysis.

Methods: A surgical specimen obtained from a 68-year-old female HCC patient was analyzed using triple combination array analysis, which identified a candidate cancer-related gene of HCC. Subsequently, samples from nine cell lines and 48 HCC patients were evaluated for their methylation and expression status of the identified gene using methylation-specific polymerase chain reaction (MSP) and quantitative real-time reverse transcriptase PCR, respectively.

Results: Using the triple combination array analysis, cyclin J (CCNJ) was detected as a candidate cancer-related gene. CCNJ was located on chromosome 10q24.1 and the copy number, using SNP chip array, revealed no loss of heterozygosity. According to expression array results, The expression of CCNJ in tumor tissue decreased at two points of the expression array chip, and the decreased values of the chip were −1.3 and −2.3. CCNJ was found to be hypermethylated (methylation value 0.906, range 0-1.0) in cancer tissue compared with adjacent normal tissue (0.112) using the methylation array. Using MSP, hypermethylation of the promoter region of CCNJ was shown to occur in 37 (77.1%) tumor samples. CCNJ expression was significantly decreased in cases with methylation (p<0.0001). Furthermore, HCC patients with methylated CCNJ had a significantly worse prognosis for recurrence-free survival (p=0.0353) and tended to have a worse prognosis for overall survival than those with unmethylated CCNJ (p=0.0838).

Conclusion: The present study indicates that triple combination array analysis is an effective method to detect novel genes related to HCC. We suggest that CCNJ acts as a cancer-related gene in HCC.

Citation Format: Nao Takano, Shuji Nomoto, Mitsuhiro Hishida, Yoshikuni Inokawa, Masamichi Hayashi, Mitsuro Kanda, Yukiyasu Okamura, Yoko Nishikawa, Chie Tanaka, Daisuke Kobayashi, Suguru Yamada, Goro Nakayama, Tsutomu Fujii, Hiroyuki Sugimoto, Masahiko Koike, Michitaka Fujii, Shin Takeda, Yasuhiro Kodera. Detection of the Cyclin J (CCNJ) as a new cancer-related gene in human hepatocellular carcinoma by using a method of triple combination array analysis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4717. doi:10.1158/1538-7445.AM2014-4717
Abstract 4729: Expression of lumican is negatively associated with the risk of biochemical recurrence in human prostate cancer
Authors: Velasco Marco A. De , Hatanaka Yuji , Oki Takashi , Kura Yurie , Yamamoto Yutaka , Yoshimura Kazuhiro , Shimizu Nobutaka , Nozawa Masahiro , Yoshikawa Kazuhiro , Nishio Kazuto , Uemura Hirotsugu
Published: May 2015

We used DNA chip technology in a genetically engineered mouse model of prostate cancer, and identified lumican as a candidate biomarker in prostate cancer progression. Lumican is a small leucine-rich extracellular matrix protein that has been shown to modulate cell migration and proliferation during embryonic development, and tissue repair. Expression and distinct glycosylation patterns of lumican in several tumor and stromal tissues has been shown to play important roles in vascular invasion, differentiation, proliferation, and invasion. Our goal was to determine the clinical value of lumican as a biomarker for human prostate cancer. In order to do so, we examined gene expression in publicly available databases and confirmed the protein expression in tissue microarrays constructed from prostatectomy cases. Protein expression was examined by immunohistochemistry and expression scores were compared with clinicopathological features and disease progression. Our analysis shows that in human prostate cancer, stromal expression of lumican correlated inversely with clinicopathological features associated with disease progression. In addition, biochemical recurrence was significantly longer for patients with high expression of stromal lumican. Our findings suggest that lumican may be a strong candidate prognostic biomarker and warrants prospective validation.

Citation Format: Marco A. De Velasco, Yuji Hatanaka, Takashi Oki, Yurie Kura, Yutaka Yamamoto, Kazuhiro Yoshimura, Nobutaka Shimizu, Masahiro Nozawa, Kazuhiro Yoshikawa, Kazuto Nishio, Hirotsugu Uemura. Expression of lumican is negatively associated with the risk of biochemical recurrence in human prostate cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4729. doi:10.1158/1538-7445.AM2014-4729
Abstract 473: TRIM59, a novel prognostic marker for gastric cancer, promotes tumorigenesis via ubiquitination and degradation of the p53 tumor suppressor
Authors: Zhou Zhicheng , Wang You , Li Jian , Cao Hui , Zhu Helen H. , Gao Wei-Qiang
Published: May 2015

As a key tumor suppressor, p53 plays crucial roles in gastric cancer initiation and progression. However, the underlying mechanisms for the down-regulation of p53 in gastric cancer patients are not fully understood. Several tripartite motif (TRIM) family proteins are identified to be involved in the tumorigenesis of multiple cancer types, but the significance and biological function of a new TRIM family protein, TRIM59, in terms of whether and how it affects p53 signaling in gastric cancer are largely unknown. We analyzed multiple microarray datasets from the Oncomine database and tested the expression level of TRIM59 in gastric tumors vs. their matched normal tissues and in gastric carcinoma cell lines using quantitative-polymerase chain reactions (Q-PCR) and immunohistochemical staining. We found Expression of TRIM59 was significantly up-regulated in gastric cancer and strongly associated with poor prognosis. TRIM59 knockdown led to significantly attenuated gastric cancer cell proliferation, clone formation, migration, and in vivo xenograft tumorigenicity, whereas over-expression of TRIM59 showed the opposite effects. Mechanistically, TRIM59 interacted with p53 and enhanced ubiquitination and degradation of the p53 protein. In addition, elevated expression of TRIM59 correlated with down-regulation of p53 target genes in human gastric tumors. In summary, TRIM59 is a novel prognostic marker for gastric cancer and promotes tumorigenesis through facilitating p53 ubiquitination and degradation.

Citation Format: Zhicheng Zhou, You Wang, Jian Li, Hui Cao, Helen He Zhu, Wei-Qiang Gao. TRIM59, a novel prognostic marker for gastric cancer, promotes tumorigenesis via ubiquitination and degradation of the p53 tumor suppressor. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 473. doi:10.1158/1538-7445.AM2014-473
Abstract 4734: Activation of yap1 is significantly associated with poor prognosis and cetuximab resistance in colorectal cancer patients
Authors: Lee Keun-Wook , Lee Sung Sook , Kim Sang-Bae , Park Yun-Yong , Sohn Bo Hwa , Lee Hyun-Sung , Lee Ju-Seog
Published: May 2015

Purpose: To investigate the clinical significance of activation of YAP1, a newly identified oncogene in colorectal cancer (CRC).

Patients and Methods: A gene expression signature reflecting YAP1 activation was identified in CRC cells, and CRC patients were stratified into two groups according to this signature: activated YAP1 CRC (AYCC) or inactivated YAP1 CRC (IYCC). Stratified patients in six test cohorts were evaluated to determine the effect of the signature on CRC prognosis and response to treatment with cetuximab.

Results: The activated YAP1 signature was associated with poor prognosis for CRC in four independent patient cohorts with stage I-III disease (total n = 1,028). In a multivariate analysis, the impact of the YAP1 signature on the disease-free survival rate was independent of other clinical variables [hazard ratio (HR), 1.63; 95% confidence interval (CI), 1.25-2.13; P < .001]. In patients with stage IV CRC, AYCC patients had a poorer disease control rate and progression-free survival duration after cetuximab-based monotherapy than did IYCC patients (HR, 1.82; 95% CI, 1.05-3.16; P = .03). In multivariate analysis, the effect of YAP1 activation on PFS was independent of KRAS mutation status and cetuximab-based therapy was effective only in IYCC patients without KRAS mutations.

Conclusion: Activation of YAP1 is highly associated with poor prognosis for CRC and may be useful in identifying patients with CRC resistant to treatment with cetuximab.

Citation Format: Keun-Wook Lee, Sung Sook Lee, Sang-Bae Kim, Yun-Yong Park, Bo Hwa Sohn, Hyun-Sung Lee, Ju-Seog Lee. Activation of yap1 is significantly associated with poor prognosis and cetuximab resistance in colorectal cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4734. doi:10.1158/1538-7445.AM2014-4734
Abstract 4746: Selective inhibition of mutant IDH1 via small molecule binding to the dimer interface
Authors: Deng Gejing , Licht Stuart , Shen Junqing , Yin Ming , McManus Jessica , Gee Patricia , He Tim , Gao Giang , Zhang Bailin , Mathieu Magali , Rak Alexey , Bedel Olivier , Shi Chaomei , Gross Stefan , Hoffmann Dietmar , Rooney Eamonn , Vassort Aurelie , Englaro Walter , Li Yi , Wiederschain Dmitri , Patel Vinod , Adrian Francisco , Cheng Hong
Published: May 2015

Isocitrate dehydrogenases (IDHs) catalyze the oxidative decarboxylation of isocitric acid to α-ketoglutaric acid (αKG). Point mutations in IDH1 and IDH2 confer a neomorphic enzymatic activity: the reduction of αKG to D-2-hydroxyglutaric acid (2HG), which acts as an oncometabolite by inducing hypermethylation of histones and DNA. IDH1 mutations (predominantly R132H) have been found in 50-80% of grade II gliomas. To find potential IDH1-targeted therapeutics, we conducted a full small-molecule HTS and hit validation campaign using recombinant R132H IDH1. This campaign identified Compound 1, which belongs to a phenol-azole series, as a potent and selective inhibitor of mutant IDH1. We investigated the mode of inhibition of Compound 1 and a published IDH1 mutant inhibitor with a different chemical scaffold (Compound 2). Steady-state kinetics and biophysical studies showed that both inhibitors reversibly interact with both free and substrate-bound enzyme at an allosteric site. A crystal structure of Compound 1 complexed with R132H demonstrates binding of the inhibitor at the dimer interface. A competitive binding study showed that Compound 2 competes with Compound 1, suggesting that Compound 2 also binds at the dimer interface. This is contrary to a previously published statement that Compound 2 is a competitive inhibitor with respect to αKG. In cancer cell lines engineered to produce high levels of 2HG, both inhibitors penetrate cells efficiently and inhibit 2HG production with minimal cell toxicity. This study indicates that the dimer interface pocket is a druggable binding site for IDH1 inhibitors, and suggests that Compound 1 is a promising new starting point for future structure-based drug discovery efforts.

Citation Format: Gejing Deng, Stuart Licht, Junqing Shen, Ming Yin, Jessica McManus, Patricia Gee, Tim He, Giang Gao, Bailin Zhang, Magali Mathieu, Alexey Rak, Olivier Bedel, Chaomei Shi, Stefan Gross, Dietmar Hoffmann, Eamonn Rooney, Aurelie Vassort, Walter Englaro, Yi Li, Dmitri Wiederschain, Vinod Patel, Francisco Adrian, Hong Cheng. Selective inhibition of mutant IDH1 via small molecule binding to the dimer interface. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4746. doi:10.1158/1538-7445.AM2014-4746
Abstract 4758: Disruption of CRAF-mediated MEK activation is required for effective MEK inhibition in KRAS mutant tumors
Authors: Lito Piro , Saborowski Anna , Yue Jingyin , Solomon Martha , Joseph Eric , Fellman Christof , Ohara Kazuhiro , Morikami Kenji , Miura Takaaki , Lucas Christine , Ishii Nobuya , Lowe Scott , Rosen Neal
Published: May 2015

MEK inhibitors are clinically active in BRAF V600E melanomas, but only marginally so in KRAS-mutant tumors. While MEK inhibitor treatment resulted in sustained inhibition of ERK phopshorylation in BRAF V600E tumors, it was associated with a rebound in ERK phosphorylation after prolonged exposure in KRAS mutant tumors. To understand this phenomenon, we performed an RNAi screen in a KRAS-mutant model and found that CRAF knockdown enhanced the antiproliferative effects of MEK inhibition both in vivo and in vitro. In agreement with previous findings, MEK inhibitor treatment in KRAS mutant cells resulted in CRAF reactivation and induction of MEK phosphorylation. However, MEK activated by CRAF was less susceptible to traditional MEK inhibitors, such as PD0325901, compared to when activated by BRAF V600E. Consistent with this observation, the rebound in ERK phosphorylation following MEK inhibitor treatment in KRAS mutant tumors was dependent on intact CRAF expression. Furthermore, traditional MEK inhibitors induced RAF-MEK complexes in KRAS mutant models, and disrupting these complexes enhanced the inhibition of CRAF-dependent ERK signaling. In an effort to identify more effective compounds we found that newer MEK inhibitors, such as trametinib and CH5126766, do not only target the catalytic activity of MEK, but in addition, they also impair its reactivation by CRAF. This occurs through two distinct mechanisms: trametinib disrupts RAF-MEK complex formation, as evidenced by co-immunoprecipitation of endogenous proteins and surface plasmon resonance assays with purified enzymes. Instead, CH512676 induces the interaction of MEK with RAF, yet it prevents the phosphorylation of MEK in this complex. To elucidate the mechanism responsible for the latter property, we determined the ternary structure of CH512676-bound MEK1. CH5126766 interacts with Asn 221 and Ser 222 residues in MEK1 to displace the activation segment and disrupt RAF-mediated phosphorylation. These data provide a blueprint for developing better MEK inhibitors that effectively inhibit ERK signaling in KRAS mutant tumors.

Citation Format: Piro Lito, Anna Saborowski, Jingyin Yue, Martha Solomon, Eric Joseph, Christof Fellman, Kazuhiro Ohara, Kenji Morikami, Takaaki Miura, Christine Lucas, Nobuya Ishii, Scott Lowe, Neal Rosen. Disruption of CRAF-mediated MEK activation is required for effective MEK inhibition in KRAS mutant tumors. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4758. doi:10.1158/1538-7445.AM2014-4758
Abstract 4765: Drug synergies observed for antibody and toxin components of SAR3419 ADC contribute to overall conjugate efficacy and can be combination drug or tumor cell line dependent
Authors: Rickles Richard J. , Giordano Thomas P. , Cotard Shakira F. , Grenier Jill M. , Romanelli Angela , Cai Ti
Published: May 2015

SAR3419, a novel antibody-drug conjugate (ADC), is composed of a humanized monoclonal IgG1 anti-CD19 antibody (huB4) attached by a cleavable linker to DM4, a derivative of cytotoxic agent maytansine (an inhibitor of microtubule polymerization). CD19 expression is restricted to B-cells and treatment of CD19-positive human lymphoma cells with SAR3419 can result in cell cycle arrest followed by apoptosis. As traditional monotherapies can rarely treat cancer effectively, particularly in non-solid tumours, a combination high throughput screen (cHTS) was performed where SAR3419 was combined with two hundred potential enhancer compounds across a panel of twenty cell lines using a 6x6 combination dose response matrix. The enhancer compounds compose a library of approved, emerging oncology drugs, and well-defined molecular probes. Robust synergies were observed between SAR3419 and PI3K inhibitors and examined further. Target/pathway inhibitor redundancy for the class of compounds served to validate PI3K inhibition as important for combination synergy. Surprisingly, for several of the cell lines, the molecular mechanism important for SAR3419-dependent combination activity is distinct. For example, for the Daudi and REC-1 tumor cell lines, PI3K inhibitor synergy can be attributed to the toxin DM4. In contrast for SU-DHL4 and SU-DHL6 cell lines, little combination activity is observed with toxin alone while the conjugate is strongly synergistic. By examining three-way combination activities (PI3K inhibitor x DM4 x huB4) using high resolution matrices, we show that the anti-CD19 antibody huB4 contributes to the synergy in the SU-DHL6 cell line and linkage of DM4 to huB4 (SAR3419) shifts both the potency and maximum activity for PI3K inhibitor combinations. The results demonstrate that the antibody component of an ADC can contribute to combination activity for specific drug pairings in a drug and cell-type specific manner and has wide ranging implications with respect to combination drug design.

Citation Format: Richard J. Rickles, Thomas P. Giordano, Shakira F. Cotard, Jill M. Grenier, Angela Romanelli, Ti Cai. Drug synergies observed for antibody and toxin components of SAR3419 ADC contribute to overall conjugate efficacy and can be combination drug or tumor cell line dependent. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4765. doi:10.1158/1538-7445.AM2014-4765
Abstract 4775: PI3K-p110β is required for leukemic transformation and HSC depletion in the absence of Pten
Authors: Yuzugullu Haluk , Baitsch Lukas , Steiner Allison , Clayton Linda K. , Gritsman Kira , Zhao Jean J.
Published: May 2015

The tumor suppressor PTEN, which antagonizes PI3K signaling, plays critical roles in cell cycle, cell migration and stem cell self-renewal. PTEN is frequently mutated or inactivated in T-cell acute lymphoblastic leukemia (T-ALL) and acute myelogenous leukemia (AML). In mice, HSC-specific deletion of Pten leads to myeloproliferative phenotype, transplantable leukemia, and increased cycling of hematopoietic stem cells (HSCs), leading to their depletion. Rapamycin, an mTOR inhibitor, rescues this phenotype. Thus, it is likely that PI3K itself plays an important role in Pten-deficient HSCs and leukemic cells, but the specific contributions of the PI3K isoforms are poorly understood. We demonstrate that genetic ablation or pharmacological inhibition of PI3K-p110β prevents myeloid neoplasia and prolongs survival. Importantly, p110β ablation rescues HSCs in the bone marrow (BM), and reinstates long-term hematopoietic reconstitution. Moreover, Pten deficiency leads to aberrant expression of Cxcl12 in the BM and spleen, enabling the generation of an alternative splenic niche. Inactivation of p110β rescues retention of HSCs, and perturbs the splenic niche by interfering with the Cxcl12-CXCR4 axis. This suggests that selective inhibition of p110β offers an alternative therapeutic strategy in PTEN deficient leukemias while restoring untransformed HSCs to their proper niche.

Citation Format: Haluk Yuzugullu, Lukas Baitsch, Allison Steiner, Linda K. Clayton, Kira Gritsman, Jean J. Zhao. PI3K-p110β is required for leukemic transformation and HSC depletion in the absence of Pten. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4775. doi:10.1158/1538-7445.AM2014-4775
Abstract 4780: The effects of the global loss of DNA methylation on the functional organization of the epigenome
Authors: Lay Fides D. , Liu Yaping , Kelly Theresa K. , Witt Heather , Blattler Adam , Farnham Peggy J. , Berman Ben P. , Jones Peter A.
Published: May 2015

Aberrant epigenetic changes which include alterations in DNA methylation, nucleosome positioning and histone modifications have been studied extensively in the context of tumorigenesis. The role of DNA methylation in the organization of genome-wide chromatin structure and its interaction with other epigenetic modifiers in cancer, however, remains unclear. Here, we use the Nucleosome Occupancy and Methylation whole-genome sequencing (NOMe-seq) assay to compare a colorectal cancer cell line, HCT116, with its derivatives that carry homozygous deletions of DNMT3B and are hypomorphic for DNMT1, DKO1, to examine the role of DNA methylation in the functional organization of the cancer epigenome. Upon the global loss of DNA methylation, we observe a widespread chromatin remodeling where specifically, DKO1 cells gain nucleosome depleted regions (NDRs) and/or well-phased nucleosome that are not present in the parent HCT116 cells. Interestingly, this genome-wide nucleosome repositioning in DKO1 cells is accompanied by significant histone modification changes and low level expression increase only in CpG islands (CGI), where promoters may gain permissive histone marks H2A.Z, H3K4me3 and H3K4me1 as well as the polycomb repressive mark, H3K27me3. Non-CGI promoters on the other hand, do not gain permissive or repressive histone modifications despite dramatic nucleosome reorganization, which suggests that distinct mechanisms regulate the equilibrium between DNA methylation, nucleosome and gene expression in CGI and non-CGI promoters. Taken together, our study demonstrates the cross-talk between epigenetic mechanisms wherein DNA methylation may directly control the organization of chromatin architecture.

Citation Format: Fides D. Lay, Yaping Liu, Theresa K. Kelly, Heather Witt, Adam Blattler, Peggy J. Farnham, Ben P. Berman, Peter A. Jones. The effects of the global loss of DNA methylation on the functional organization of the epigenome. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4780. doi:10.1158/1538-7445.AM2014-4780
Abstract 4781: Integrated epigenomic profiling reveals widespread demethylation in melanoma and reveals CSF-1 receptor as an aberrant regulator of malignant growth and invasion
Authors: Mo Yongkai , Giricz Orsolya , Hu Caroline H. , Dahlman Kimberly B. , Bhattacharyya Sanchari , Nguyen Hoa , Matusow Bernice , Bhagat Tushar , Shellooe Rafe , Burton Elizabeth , Tsai James , Zhang Chao , Habets Gaston , Shyr Yu , Greally John , Yu Yiting , Bollag Gideon E. , Stanley Richard , Trent Jeffrey , Kenny Paraic A. , West Brian L. , Sosman Jeffrey , Verma Amit K.
Published: May 2015

Epigenetic alterations can direct carcinogenesis by leading to transcriptional changes and inducing genomic instability. We analyzed the methylome of malignant melanoma and observed widespread loss of DNA methylation that was found to preferentially occur outside of CpG islands. Demethylation was seen to occur early during carcinogenesis, was independent of mutational status and correlated with genomic instability. Parallel transcriptomic analyses revealed that various immune and cancer associated pathways were overexpressed and were associated with promoter demethylation. The CSF1-receptor (CSF1R) was aberrantly overexpressed and hypomethylated in nearly all cases and was strikingly expressed via an aberrant upstream promoter in 10% of melanomas. shRNA mediated knockdown and inhibition of CSF1R kinase via a clinically relevant inhibitor, PLX3397, led to decreased 3D growth and invasiveness. Co-inhibition of CSF1R and BRAF resulted in synergistic blockade of BRAF-mutant melanoma xenograft growth. Thus, widespread epigenetic changes are seen in melanoma and CSF1R is a potential therapeutic target in this disease.

Citation Format: Yongkai Mo, Orsolya Giricz, Caroline H. Hu, Kimberly B. Dahlman, Sanchari Bhattacharyya, Hoa Nguyen, Bernice Matusow, Tushar Bhagat, Rafe Shellooe, Elizabeth Burton, James Tsai, Chao Zhang, Gaston Habets, Yu Shyr, John Greally, Yiting Yu, Gideon E. Bollag, Richard Stanley, Jeffrey Trent, Paraic A. Kenny, Brian L. West, Jeffrey Sosman, Amit K. Verma. Integrated epigenomic profiling reveals widespread demethylation in melanoma and reveals CSF-1 receptor as an aberrant regulator of malignant growth and invasion. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4781. doi:10.1158/1538-7445.AM2014-4781