bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4158: Identification of somatic mutations in EGFR/KRAS/ALK-negative lung adenocarcinoma from never smokers
Authors: Kim Han Sang , Ahn Jin Woo , Yoon Jung-Ki , Han Soo Min , Jang Hoon , Eun Sungho , Shim Hyo Sup , Kim Hyun Jung , Kim Dae Joon , Lee Jin G. , Lee Chang Young , Bae Mi Kyung , Chung Kyung Young , Kim Eun Young , Jung Ji Y. , Kim Se Kyu , Chang Joon , Kim Hye Ryun , Kim Joo Hang , Lee Ji Hyun , Bang Duhee , Cho Byoung Chul
Published: May 2015

Background: Lung adenocarcinoma is a highly heterogeneous disease with various etiologies, prognoses, and responses to therapy. Although genome-scale characterization of lung adenocarcinoma has been performed, somatic mutation analysis of never-smoker EGFR/KRAS/ALK-negative lung adenocarcinoma has not been comprehensively conducted.

Methods: Patients with lung adenocarcinoma were eligible if they were never smoker without EGFR/KRAS mutation and ALK rearrangement. Whole exome sequencing was performed using the Illumina HiSeq 2,000 platform. Candidate loci identified in whole exome sequencing were validated using hybridization capture method and Sanger sequencing.

Results: We present an analysis of whole exome sequencing data from 16 EGFR/KRAS/ALK-negative lung adenocarcinomas with additional 54 tumors in two independent cohort sets. We identified 27 genes potentially implicated in the pathogenesis of lung adenocarcinoma. In addition to targetable PI3K/mTOR signaling (TSC1, PIK3CA, AKT2) and receptor tyrosine kinase (RTK) signaling (ERBB4) genes, some of the genes including SETD2 and PBRM1 (chromatin remodeling), CHEK2 and CDC27 (cell cycle), CUL3 and SOD2 (oxidative stress), and CSMD3 and TFG (immune response) have not been previously highlighted in lung adenocarcinomas. In total (N=70), TP53 is the most frequently altered gene (11%), followed by SETD2 (6%), CSMD3 (6%), ERBB2 (6%), and CDH10 (4%). In pathway analysis, majority of altered genes were involved in cell cycle/DNA repair (P<0.001), and cAMP-mediated protein kinase A signaling (P<0.001).

Conclusions: Genomic makeup of EGFR/KRAS/ALK-negative adenocarcinomas in never-smokers was remarkably diverse. Impairment in cell cycle/DNA repair may be involved in tumorigenesis and therapeutic targets in lung adenocarcinoma.

Note: This abstract was not presented at the meeting.

Citation Format: Han Sang Kim, Jin Woo Ahn, Jung-Ki Yoon, Soo Min Han, Hoon Jang, Sungho Eun, Hyo Sup Shim, Hyun Jung Kim, Dae Joon Kim, Jin Gu Lee, Chang Young Lee, Mi Kyung Bae, Kyung Young Chung, Eun Young Kim, Ji Ye Jung, Se Kyu Kim, Joon Chang, Hye Ryun Kim, Joo Hang Kim, Ji Hyun Lee, Duhee Bang, Byoung Chul Cho. Identification of somatic mutations in EGFR/KRAS/ALK-negative lung adenocarcinoma from never smokers. [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 4158. doi:10.1158/1538-7445.AM2014-4158
Abstract 4162: Circulating C-reactive protein and lung cancer risk: A nested case-control study in the Southern Community Cohort Study
Authors: Wu Jie , Zheng Wei , Wen Wanqing , Hargreaves Margaret K. , Blot William J. , Cai Qiuyin
Published: May 2015

Lung cancer is the leading cause of cancer deaths in the United States and many other countries. Cumulative evidence suggests that chronic inflammation may play an etiologic role in lung carcinogenesis. C-reactive protein (CRP), a circulating biomarker of systemic inflammation, has been associated with increased lung cancer risk in several retrospective and a few prospective studies. However, prior studies included only a small number of African Americans with lung cancer, a group with higher lung cancer incidence rates than other ethnic or racial groups. We conducted a nested case-control study to evaluate the association of pre-diagnostic circulating levels of CRP with subsequent risk of lung cancer using data and biological samples collected in the Southern Community Cohort Study (SCCS), a prospective cohort study, in which approximately 86,000 men and women aged 40-79 years, two-thirds of whom are African American, were recruited from 2002 to 2009. Plasma CRP levels in 396 incident lung cancer cases (258 African Americans and 130 Whites) were compared with 784 matched controls. Overall, baseline plasma CRP levels were 29.4% higher among participants who subsequently developed lung cancer than among those who remained free of the disease (geometric mean: 4.4 µg/ml vs. 3.4 µg/ml; P = 0.0015). This difference was seen primarily among African Americans (geometric mean: 4.1 µg/ml vs. 3.0 µg/ml; P=0.0026) and not among Whites (geometric mean: 5.0 µg/ml vs. 4.5 µg/ml; P=0.3613). In both African Americans and Whites, elevated risk of lung cancer was associated with high circulating levels of CRP. However, the association was only statistically significant among African Americans; adjusted odds ratios (ORs) for developing lung cancer increased from 1.0 to 1.46 (95%CI: 0.89-2.40); 1.52 (95%CI: 0.88-2.61), and 2.95 (95%CI: 1.77-4.93) with increasing quartiles of plasma CRP (P-trend = 0.0018). The findings from this study suggest that circulating CRP level is positively associated with prospective lung cancer risk among African Americans.

Citation Format: Jie Wu, Wei Zheng, Wanqing Wen, Margaret K. Hargreaves, William J. Blot, Qiuyin Cai. Circulating C-reactive protein and lung cancer risk: A nested case-control study in the Southern Community Cohort Study. [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 4162. doi:10.1158/1538-7445.AM2014-4162
Abstract 4170: Oncogenic activation of LMO2 by EGFR signaling regulates STAT3 phosphorylation and transcriptional activity in human glioma
Authors: Fu Jun , Koul Dimpy , Wu Shaofang , Zheng Siyuan , Verhaak Roel G. , Yung W.K
Published: May 2015

Glioblastoma multiforme (GBM) is a highly heterogeneous disease characterized by multiple genetic abnormalities. Amplification and mutation of the epidermal growth factor receptor (EGFR) gene represent signature genetic abnormalities occurred in over 60% of GBM. The successful application of EGFR-targeted therapy for the treatment of GBM is thus dependent on a thorough understanding of the intricate signal pathways cross-linkage that underlies the pathobiology of this disease. The LIM-domain only factor 2 (LMO2) gene is a crucial oncogene involved in hematopoietic development and leukemogenesis. Herein, we showed, from the TCGA GBM data, that LMO2 is highly expressed in EGFR-aberrant GBM samples and might serve as a downstream effector of EGFR signaling in GBM. Depletion of LMO2 expression impairs the maintenance of glioma cell proliferation in vitro and in vivo. In addition, EGFR signaling induces LMO2 nuclear translocation and enables LMO2 to form a triplex with EGFR and Signal transducer and activator of transcription 3 (STAT3). More importantly, LMO2 serves as a bridging molecule to promote the formation of EGFR/STAT3 complex and to enhance EGFR-mediated STAT3 Y705 phosphorylation upon EGF stimulation, which in turn up-regulates the expression of STAT3-responsive genes involved in the regulation of cell proliferation and tumor development. In conclusion, these findings reveal that LMO2 might serve as a critical effector of EGFR signaling in human glioma and highlight the essential function of LMO2 in EGF-stimulated STAT3 activation and tumorigenesis.

Citation Format: Jun Fu, Dimpy Koul, Shaofang Wu, Siyuan Zheng, Roel G. Verhaak, W.K Yung. Oncogenic activation of LMO2 by EGFR signaling regulates STAT3 phosphorylation and transcriptional activity in human glioma. [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 4170. doi:10.1158/1538-7445.AM2014-4170
Abstract 4176: Identifying analytes associated with poor prognosis in CRC
Authors: French Christi Lynn , Weaver Alissa
Published: May 2015

Cancer patients with undetectable micrometastases at the time of diagnosis are in danger of subsequent metastatic growth that worsens their outcome. Therefore, identification of these patients with latent metastases could reduce morbidity and mortality by helping physicians to choose whether to treat with an adjuvant therapy. In colorectal carcinoma (CRC), 20-30% of stage II patients will have a recurrence at a distant site after removal of the primary tumor. In order for cancer cells to metastasize, they must switch to an “invasive phenotype”. To identify molecular changes in cancer that drive invasion, our laboratory previously built a molecular network model of invasive subcellular structures termed invadopodia. Centrality and random walk analyses identified candidate signaling hubs that may control invadopodia formation or activity. To identify which of these hubs control invasion in specific cancers, we are mining reverse phase protein array (RPPA) data from publicly available human tumor datasets. In this study, we focused on analysis of RPPA data from TCGA in CRC patients, segregated by poor prognosis and good prognosis. We tested three different definitions of poor prognosis: recurrence, survival, and TNM status at diagnosis. We found several promising analytes that were significantly associated with each poor prognosis definition, and recurrence provided the best separation of good vs. poor prognosis. Interestingly, many of the analytes significantly associated with survival are involved in apoptosis. Additionally, we refined the analysis by using a subset of patients (stage I-II), which resulted in even better separation of good vs. poor prognosis. We identified several analytes that are already significantly associated with the invadopodia network (src kinase and PI 3-kinase). We also identified some potential new invadopodia regulators, such as Cox2. This method has potential both to improve CRC survival by identifying patients that need adjuvant therapy as well as identify novel drivers of invadopodia.

Citation Format: Christi Lynn French, Alissa Weaver. Identifying analytes associated with poor prognosis in CRC. [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 4176. doi:10.1158/1538-7445.AM2014-4176
Abstract 4181: Accurate and fast detection and comparison of larger clinically relevant insertions and deletions
Authors: Hein Anne-Mette K. , Dekker Patrick , Joecker Anika , Boysen Cecilie , Thomson Naomi , Oester Bodil , Arens Anne , Knudsen Bjarne , Knudsen Thomas , Forsberg Roald
Published: May 2015

Larger somatic insertions and deletions in tumor samples are often of significant clinical impact. Many of them are cancer driver mutations and play an important role in drug treatment1.

Detecting the accurate breakpoints of larger insertions and deletions is often problematic and inaccurate. Furthermore, due to ambiguous positioning of them in the human genome, they are hard to compare with known deletions and insertions in publicly available databases.

Here we present a complete analysis workflow to accurate identify, filter and annotate larger insertions and deletions with information from publicly available databases such as COSMIC. We apply our pipeline on publicly available tumor/normal matched pair data from a patient with Massive Acinic Cell Carcinoma (A. Nichols et al. (2013) Case Reports in Oncological Medicine, Article ID 270362), which has not been investigated for larger insertions and deletions before. We show that we can identify insertions and deletions, which should be reported as part of the list of somatic variants, the authors have presented. Moreover, we will point out potential challenges and how to solve them while comparing own results to data in databases.

Citation Format: Anne-Mette K. Hein, Patrick Dekker, Anika Joecker, Cecilie Boysen, Naomi Thomson, Bodil Oester, Anne Arens, Bjarne Knudsen, Thomas Knudsen, Roald Forsberg. Accurate and fast detection and comparison of larger clinically relevant insertions and deletions. [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 4181. doi:10.1158/1538-7445.AM2014-4181
Abstract 4187: Splicing factors ESRP1/ESRP2 as regulators of endocrine resistance in breast cancer
Authors: Gokmen-Polar Yesim , Nellamraju Yaseswini , Gu Xiaoping , Janga Sarath C. , Badve Sunil S.
Published: May 2015

Deregulation of alternative splicing (AS) has been implicated in the development of drug resistance, tumor progression and metastasis. RNA-binding proteins (RBPs) as controllers of AS may act as master regulators of cellular behavior. Among the RBPs, we previously have shown that the expression of epithelial splicing regulatory proteins 1 and 2 (ESRP1 & ESRP2) have significantly elevated in cases with high Oncotype DX scores and in ERα-positive cells with acquired tamoxifen resistance (MCF-7/ LCC2) (SABCS 2013). This study seeks to identify genes and their splice variants that are regulated by ESRP1 & ESRP2 and could cause development of endocrine resistance. To identify alternatively spliced genes by ESRP1 & ESRP2 in endocrine resistance, we have performed a four step bioinformatics approach as follows: 1) Motif analysis to identify genes targeted by ESRP1 & ESRP2, 2) Comparison of differentially expressed genes in tamoxifen treated samples (GSE17700 and GSE17705) with genes targeted by ESRP1 & ESRP2 identified above, 3) Protein-protein interaction from BIOGRID and 4) survival data using publically available datasets (KM Plotter). Motif analysis identified 15,450 genes targeted both by ESRP1 (P<3.39e-05) and ESRP2 (P< 5.53e-05). 2212 of these genes were differentially expressed in tamoxifen treated cases. This set of genes was further categorized based on the ESRP1 & ESRP2 enhancer/ suppressor motifs ratio resulting in 1260 genes. Further collective analysis of survival data from KM plotter (P<0.001) and number of protein-protein interactions (>50) narrowed down 79 candidate genes that are associated with tamoxifen resistance. Some of the genes (AURKA, ERRB3, ESR2, EGFR, and MDM2) have been previously implicated in the de novo or acquired resistance to tamoxifen. Validation of splicing variants of these genes in human tumors is ongoing. Elevated expression of ESRP1 & ESRP2 and its target genes play an important role in the development of tamoxifen resistance and recurrence of ER+ breast cancer. ESRP1 & ESRP2 by affecting splicing have the potential to induce tamoxifen resistance via multiple genes and pathways. Targeting ESRP1 & ESRP2 may offer novel avenues for combating endocrine-resistance in breast cancer.

Citation Format: Yesim Gokmen-Polar, Yaseswini Nellamraju, Xiaoping Gu, Sarath C. Janga, Sunil S. Badve. Splicing factors ESRP1/ESRP2 as regulators of endocrine resistance in 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 4187. doi:10.1158/1538-7445.AM2014-4187
Abstract 4189: Discovery of a novel carcinoma-associated EF hand containing protein by mining the dark matter of the human proteome
Authors: Delgado Ana Paula , Brandao Pamela , Hamid Sheilin , Ven Wim Van de , Narayanan Ramaswamy
Published: May 2015

The human genome contains thousands of novel and uncharacterized proteins. These unknown genes are difficult to mine due to lack of adequate information. Hence, they remain largely untapped. Reasoning that new druggable targets for cancer can be discovered from these uncharacterized proteins, we have developed a protocol to mine these proteins for cancer relevance. Using various cancer-related databases, we have developed a database of uncharacterized ORF proteins. Numerous ORFs from this database offer druggable targets. To develop a proof of concept for mining these targets, we have chosen one target, C1ORF87 for detailed characterization. The C1ORF87 was analyzed using bioinformatics approaches for mRNA and protein expression, SNPs, LOH, protein motifs and domains and for gene network interactions. We have identified the C1ORF87 gene as a calcium binding protein with an EF-Hand motif. The EF-hand motifs are present in numerous calcium binding proteins, which exert a complex function in tumor growth and suppression. We show that the C1ORF87 gene is down-regulated in the lung and breast carcinomas and up-regulated in the liver carcinomas. Hematopoietic and neuronal tumors were largely negative. Its expression in select normal tissues suggests that the C1ORF87 may be a putative druggable gene. Our results suggest that the C1ORF 87 gene may play a significant role in specific carcinomas. We further demonstrate that the dark matter of the human proteome can be readily mined to predict putative cancer targets. Detailed characterization of the C1ORF87 gene will be presented.

Citation Format: Ana Paula Delgado, Pamela Brandao, Sheilin Hamid, Wim Van de Ven, Ramaswamy Narayanan. Discovery of a novel carcinoma-associated EF hand containing protein by mining the dark matter of the human proteome. [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 4189. doi:10.1158/1538-7445.AM2014-4189
Abstract 4195: Peroxisome proliferator-activated receptor α is regulated by E2F1-induced miR-19a in a feedback loop in glioma progression
Authors: Zhang Junxia , Shi Yan , Wang Yingyi , Luan Wenkang , You Yongping
Published: May 2015

The peroxisome proliferator-activated receptor α (PPARα) is a nuclear receptor, which belongs to the super-family of steroid hormone receptor. To date, the biologic function of PPARα in human cancers is still controversial and needs further investigation. In our study, we explored the clinical feature, biological function and potential mechanism of PPARα in glioma. PPARα expression was significant lower in high grade tissues than low grade tissues in whole genome gene profiling of 158 glioma tissues. PPARα activation using fenofibrate inhibited glioma cells proliferation, invasion and aerobic glycolysis. Further, PPARα was a direct target of miR-19a and miR-19a was modulated by E2F1 at transcriptional level. Moreover, PPARα activation repressed E2F1 activation by modulating transcriptional complexes formed with E2F1 and pRB proteins. Finally, we confirmed PPARα/E2F1/ miR-19a feedback loop in vivo using nude mouse glioma xenograft and human glioma tissues. To our knowledge, it is first time to demonstrate that PPARα regulates glioma development by E2F1-induced miR-19a in a feedback loop and provide important clues for understanding the key roles of PPARα functional network in glioma.

Note: This abstract was not presented at the meeting.

Citation Format: Junxia Zhang, Yan Shi, Yingyi Wang, Wenkang Luan, Yongping You. Peroxisome proliferator-activated receptor α is regulated by E2F1-induced miR-19a in a feedback loop in glioma progression. [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 4195. doi:10.1158/1538-7445.AM2014-4195
Abstract 420: Tet family proteins and 5-hydroxymethylcytosine in esophageal squamous cell carcinoma
Authors: Murata Asuka , Baba Yoshifumi , Karashima Ryuichi , Ida Satoshi , Imamura Yu , Ishimoto Takatsugu , Iwagami Shiro , Sakamoto Yasuo , Miyamoto Yuji , Yoshida Naoya , Baba Hideo
Published: May 2015

Objective: DNA methylation is a fundamental epigenetic process that modulates gene expression. Cancer cells exhibit two types of DNA methylation alterations: global DNA hypomethylation and site-specific CpG island promoter hypermethylation. Although the base 5-methylcytosine (5mc) and the DNA methyltransferase (DNMT) family of enzymes that catalyze DNA methylation are well-studied, the mechanisms of DNA demethylation are poorly understood. The Ten-eleven translocation (Tet) family of enzymes (Tet1, Tet2, and Tet3) has been implicated in DNA demethylation. These enzymes possess the dioxygenase activity and convert 5-mC to 5-hydroxymethylcytosine (5-hmC). Frequent Tet mutational inactivation or reduction has been reported to associate with decreased 5-hmC levels in various types of myeloid leukemia and melanoma. However, the status of 5hmC and Tet expression in esophageal squamous cell carcinoma (ESCC) remains still unknown.

Method: We analyzed the 5-hmC status in DNA from ESCC frozen tissues utilizing the ELISA test, and 5-hmC expression was evaluated by immunohistochemistry. We next examined the expression levels of Tet family mRNA in both normal and tumor tissues by RT-qPCR.

Results: 5-hmC level was significantly lower in ESCC than in paired normal tissue in ELISA test (n=33, P<0.0001). The similar result was observed in immunohistochemistry. We found that the Tet2 mRNA expression was significantly lower in ESCCs than paired normal tissues (n=32, P<0.0001). The expression level of Tet2 mRNA was significantly associated with the 5hmC status (n=31, r=0.54, P=0.0085). Finally, 5-hmC status did not correlate with overall survival (P = 0.65).

Conclusion: The 5hmC expression was decreased in ESCC tissues, and was associated with Tet2 expression level, suggesting that 5hmC and Tet2 might play a crucial role in the development of ESCCs.

Citation Format: Asuka Murata, Yoshifumi Baba, Ryuichi Karashima, Satoshi Ida, Yu Imamura, Takatsugu Ishimoto, Shiro Iwagami, Yasuo Sakamoto, Yuji Miyamoto, Naoya Yoshida, Hideo Baba. Tet family proteins and 5-hydroxymethylcytosine in esophageal squamous cell 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 420. doi:10.1158/1538-7445.AM2014-420
Abstract 4203: A transducer module consisting of Toll-like receptor 9 and Bruton's tyrosine kinase triggers acute myeloid leukemia blast proliferation
Authors: Oellerich Thomas , Mohr Sebastian , Bohnenberger Hanibal , Dubey Anjali , Münch Silvia , Wicht Johannes , Oellerich Mark F. , Perske Christina , Bug Gesine , Ströbel Philipp , Urlaub Henning , Serve Hubert
Published: May 2015

Acute myeloid leukemia (AML) is a genetically heterogeneous disease and mutations occur in various genes including those encoding for signaling molecules. A well characterized example is the tyrosine kinase Flt3, which harbours internal tandem duplications in about one third of AML patients that render the kinase constitutively active. AML cells often become addicted to such constitutively activated survival inducing signaling pathways that can be therapeutically exploited by using small molecule kinase inhibitors.

By screening bone marrow biopsies of AML patients for kinase expression profiles, we detected Bruton's tyrosine kinase (Btk) in AML blasts in 26 out of 30 analyzed cases. Btk acts as an oncogene in chronic lymphocytic leukemia as well as in certain B-cell lymphoma subtypes. As compared to diseases of lymphoid origin far less is known about the role of Btk in myeloid neoplasias.

Here, we show that Btk is often constitutively activated in AML and moreover, that genetic as well as pharmacological inhibition of Btk in AML cell lines as well as primary patient-derived AML cultures strongly diminishes AML cell growth. By applying quantitative mass spectrometry, we identified a signalosome consisting of Btk and Toll-like receptor 9 that is needed to constitutively transduce proliferation signals through the NFkB pathway. This newly identified TLR9/Btk signaling axis is of clinical interest as it is effectively druggable by Btk inhibitors which have so far only indications for the treatment of lymphoid malignancies.

Citation Format: Thomas Oellerich, Sebastian Mohr, Hanibal Bohnenberger, Anjali Dubey, Silvia Münch, Johannes Wicht, Mark F. Oellerich, Christina Perske, Gesine Bug, Philipp Ströbel, Henning Urlaub, Hubert Serve. A transducer module consisting of Toll-like receptor 9 and Bruton's tyrosine kinase triggers acute myeloid leukemia blast proliferation. [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 4203. doi:10.1158/1538-7445.AM2014-4203
Abstract 4217: Nuclear expression of BCL10 has a role in the regulation of cell growth and progression of pancreatic cancer through the activation of NF-κB-related signaling
Authors: Kuo Sung-Hsin , Yang Shih-Hung , Yeh Kun-Huei , Yeh Pei-Yen , Tien Yu-Wen , Lee Hsiao-Wei , Chen Li-Tzong , Cheng Ann-Lii
Published: May 2015

Background:

We previously reported that activation of TNF-α results in upregulation of NF-κB and nuclear translocation of BCL3 and BCL10 in breast cancer cell line and lymphoma cell line (J Biol Chem 2006;281:167-75, Blood 2008;112:2927-34). Furthermore, we recently demonstrated that BCL10 plays an important role in controlling the growth of cervical cancer cells through NF-κB dependent cyclin D1 regulation (Gynecol Oncol 2012;126:245-51). Since NF-κB activation has been shown to occur in the pathogenesis of pancreatic cancer, we sought to investigate whether BCL10-signaling possesses clinical significance in relation to pancreatic cancer.

Methods:

Three pancreatic cancer cell lines (PANC-1, AsPC-1, and BxPC-3) were used in this study. Cell cycle was analyzed by flow cytometry. The expression of protein and mRNA of BCL10- and NF-κB- related signaling was assessed by immunoblotting and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. The DNA-binding activity of NF-κB was determined by the luciferase assay. The expression of BCL10, and NF-κB in tumor cells from recurrent, locally advanced, and metastatic pancreatic patients who received chemotherapy was examined using immunohistochemical staining.

Results:

We transfected three pancreatic cancer cell lines with BCL10 small interfering RNA (siRNA), and discovered that the down-regulation of BCL10 inhibited the viability of these pancreatic cancer cells through the G1 arrest. BCL10 siRNA treatment inhibited the expression of p-IKKβ and p-IκB, and nuclear BCL3 translocation, and also down-regulated NF-κB activation and its downstream cell cycle proteins, c-Myc, and cyclin B1. The results of qRT-PCR showed that BCL10 siRNA treatment inhibited the mRNA expression levels of BCL10, BCL3, c-Myc, and cyclin B1. Our results reveal that patients with nuclear BCL10 expression had a worse median overall survival than those without (7.9 months versus 16.9 months, p = 0.007). Furthermore, nuclear expression of BCL10 was closely associated with nuclear BCL3 expression (p < 0.001), and NF-κB activation (p < 0.001).

Conclusions:

Our findings indicate that nuclear BCL10 plays an important role in controlling the growth and progression of pancreatic cancer cells through NF-κB-related signaling pathway.

Citation Format: Sung-Hsin Kuo, Shih-Hung Yang, Kun-Huei Yeh, Pei-Yen Yeh, Yu-Wen Tien, Hsiao-Wei Lee, Li-Tzong Chen, Ann-Lii Cheng. Nuclear expression of BCL10 has a role in the regulation of cell growth and progression of pancreatic cancer through the activation of NF-κB-related signaling. [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 4217. doi:10.1158/1538-7445.AM2014-4217
Abstract 4228: 6,7,4-Trihydroxyisoflavone, a daidzein metabolites, regulated glycogen synthase kinase 3β/β-catenin signaling and inhibited cell proliferation in MCF10DCIS.com human breast cancer cells
Authors: Park Hyun-Chang , Bao Cheng , Ko Jiwon , Lee Jaehoo , Lee Hong Jin
Published: May 2015

Recent studies have demonstrated that hepatic daidzein metabolites exerted significant activities in regulating different types of cancer including skin and colon. However, the role of daidzein metabolites on breast cancer has not been extensively studied. In this study, we found that 6,7,4′-trihydroxyisoflavone exerted the strongest inhibitory effect on the proliferation of MCF10DCIS.com human breast cancer cells among the daidzein metabolites we tested. Cell cycle at S and G2/M phase were arrested by 6,7,4-trihydroxyisoflavone, which were derived from regulating cyclins and cyclin dependent kinases (CDKs) including cyclin B, CDK1 and CDK2, as well as CDK inhibitor, p27 in a dose dependent manner. Furthermore, 6,7,4′-trihydroxyisoflavone increased the GSK3β activity through inhibiting the phosphorylation at Ser9, which was caused by the enhancement of phosphatase activity. These effects of 6,7,4′-trihydroxyisoflavone led to inhibition of the nuclear translocation of β-catenin, and regulation of cell cycle related proteins, cyclinB and CDK2. Taken together, 6,7,4′-trihydroxyisoflavone regulated GSK3β/β-catenin signaling, and arrested at S and G2/M phase in MCF10DCIS.com human breast cancer cells, and it suggests that the hepatic metabolite of daidzein, 6,7,4′-trihydroxyisoflavone, may be a potent agent in inhibiting MCF10DCIS.com human breast cancer growth.

Citation Format: Hyun-Chang Park, Cheng Bao, Jiwon Ko, Jaehoo Lee, Hong Jin Lee. 6,7,4-Trihydroxyisoflavone, a daidzein metabolites, regulated glycogen synthase kinase 3β/β-catenin signaling and inhibited cell proliferation in MCF10DCIS.com human breast cancer cells. [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 4228. doi:10.1158/1538-7445.AM2014-4228
Abstract 4230: 5α-dihydrotestosterone down-regulates HER2 and sensitizes breast cancer cells to Herceptin-induced apoptosis in vitro
Authors: Sultana Aisha , Naqvi Zulfiqar Ali , Khan Shaista , Siddiqui Anwar Ali , Lalani El-Nasir
Published: May 2015

Androgen inhibits growth of malignant breast cancer (BCa) cells by mechanisms that remain poorly defined. Here, we have investigated how androgen suppress proliferation and induces apoptosis in BCa cells. Treatment of MCF-7 cells with 1nM 5α-dihydrotestosterone (DHT) inhibited cell proliferation as well as endogenous HER2 expression, both at mRNA and protein levels, suggesting a link between androgen receptor (AR) and HER2 signaling pathways. Expression of HER2 and AR was also determined in patients with BCa. We found a survival advantage of AR expression in patients with HER2 positive BCa, whereas, loss of AR expression was associated with poor survival. This finding would suggest that AR loss may provide a growth advantage to HER2 over-expressing tumors. We showed the functional consequence of the stimulation of AR-signaling and inhibition of HER2-signaling on breast epithelial cell growth and on expression of 84 genes involved in BCa and estrogen-mediated signaling pathway. Exposure to a combination therapy of DHT and Herceptin resulted in additive anti-proliferative effect in human MCF-7 cells as compared to either agent alone. Furthermore, combined treatment of DHT with Herceptin reduces the expression of genes predominantly involved in proliferation, invasion, anti-apoptosis and cell cycle regulation.

These findings provide the key insights into how Herceptin and DHT produce anti-proliferative effect in BCa cells. In conclusion, adding DHT to Herceptin may provide a rational treatment option for BCa, not only by direct inhibition of cell proliferation, but also causing changes in the expression of many genes that are critically involved in the control of proliferation. Establishment of any interaction between AR and HER2 may improve the understanding of carcinogenesis and will provide helpful information in the development of novel preventive and therapeutic strategies.

Citation Format: Aisha Sultana, Zulfiqar Ali Naqvi, Shaista Khan, Anwar Ali Siddiqui, El-Nasir Lalani. 5α-dihydrotestosterone down-regulates HER2 and sensitizes breast cancer cells to Herceptin-induced apoptosis in vitro. [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 4230. doi:10.1158/1538-7445.AM2014-4230
Abstract 4233: Mef2C enhances 1,25-dihydroxyvitamin D3-induced monocytic differentiation of human myeloid leukemia cells by regulating C/EBPβ expression
Authors: Zheng Ruifang , Wang Xuening , Studzinski George P.
Published: May 2015

Myogenic enhancer factors 2 (Mef2) are members of the family of MADS (MCM1-agamous-deficiens-serum response factor)-box transcription factors known to have multiple roles in morphogenesis of skeletal, cardiac and smooth muscle cells. Recently, Mef2C was found to be also involved in hematopoiesis. Bone marrow cells isolated from Mef2C knockout mice demonstrated decreased monocytic differentiation in response to cytokine stimulation, whereas constitutive expression of Mef2C promoted monopoiesis. The above findings led us to examine the role of Mef2C in 1,25-dihydroxyvitamin D3 (1,25D)-induced monocytic differentiation. Human acute myeloid leukemia (AML) cell lines, HL60 and U937, were used in this study. We found that knockdown of Mef2C with small interfering RNA (siRNA) significantly decreased the expression of the monocytic marker CD14, as well as of nonspecific esterase (NSE), suggesting that Mef2C is required for 1,25D-induced monocytic differentiation in AML cells. Our laboratory has previously demonstrated that optimal monocytic differentiation induced by 1,25D requires ERK5, and that ERK5 expression shows positive correlation with Mef2C expression. Here we show that Mef2C knockdown demonstrates similar inhibition of CD14 expression as ERK5 knockdown. CCAAT-enhancer-binding proteins (C/EBPs) α and β are two major transcription factors that can directly regulate CD14 expression. Since 1,25D has been shown to increase both C/EBPα and β expression, and these two transcription factors form heterodimers to enhance CD14 expression, we used siRNA to knock down Mef2C and found decreased expression of 1,25D-induced C/EBPβ, but not C/EBPα. Similar results were obtained using an alternative method to titrate down Mef2C with dsDNA containing its DNA binding sequence. We conclude that Mef2C is a functional downstream target of ERK5, and that 1,25D-induced CD14 and NSE expression is mediated through ERK5/Mef2C activation. (Supported by T32 CA134268 training grant and R01 CA044722 both from the National Cancer Institute, NIH, and by the American Institute for Cancer Research grant 10A049)

Citation Format: Ruifang Zheng, Xuening Wang, George P. Studzinski. Mef2C enhances 1,25-dihydroxyvitamin D3-induced monocytic differentiation of human myeloid leukemia cells by regulating C/EBPβ expression. [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 4233. doi:10.1158/1538-7445.AM2014-4233
Abstract 4240: Targeting Galectin-3 to reverse tumor stemness and drug resistance
Authors: Kato Shumei , Seguin Laetitia , Franovic Aleksandra , Kato Hisashi , Camargo Maria , Desgrosellier Jay , Anand Sudarshan , Weis Sara , Shattil Sanford , Cheresh David A.
Published: May 2015

We have recently linked integrin αvβ3 expression to metastatic potential, stemness, and therapy resistance due to its capacity to couple to KRAS. This leads to the induction of RalB/TBK-1 and the activation of NFκB to shift tumor cells to a stem-like phenotype. To understand how integrin αvβ3 couples to KRAS, we generated β3 domain swap mutants containing either the intracellular or extracellular domains of β1 integrin. Each chimera partially reduced KRAS association with αvβ3, suggesting interactions on each side of the plasma membrane may contribute. Since the short β3 cytoplasmic tail has no putative KRAS binding regions, we considered Galectin-3 as a putative scaffolding partner to facilitate the αvβ3/KRAS association due to its reported binding to both KRAS and integrins. Indeed, treating cells with GCS-100, a complex polysaccharide that has the ability to bind to and block the effects of Galectin-3, decreased Galectin-3 surface expression, αvβ3 clustering, and αvβ3/KRAS association. Exposing cells to this drug also decreased tumor stem properties. Our observations provide a molecular basis for the αvβ3/KRAS/Galectin-3 complex and highlight the potential for Galectin-3 blockade as a therapeutic approach to reverse the aggressive behavior of αvβ3-expressing tumors.

Citation Format: Shumei Kato, Laetitia Seguin, Aleksandra Franovic, Hisashi Kato, Maria Camargo, Jay Desgrosellier, Sudarshan Anand, Sara Weis, Sanford Shattil, David A. Cheresh. Targeting Galectin-3 to reverse tumor stemness and drug resistance. [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 4240. doi:10.1158/1538-7445.AM2014-4240
Abstract 4242: Effects of dual inhibition of EGFR and Aurora A kinase in a lung cancer model
Authors: Bagnyukova Tetyana , Pan Xiaofan , Zhang Jiping , Egleston Brian , Astsaturov Igor , Golemis Erica , Borghaei Hossein
Published: May 2015

Lung cancer is the leading cause of cancer related death in the US. Patients with advanced disease generally have a poor prognosis with a median survival of around 10 to 12 months with standard chemotherapy. Astsaturov and colleagues (2010) employed a synthetic lethal screening method and have reported that the combined inhibition of the EGFR and Aurora A kinase pathways had synergistic effect on cancer cell viability in head and neck and colon cancer cell lines suggesting a potential benefit of such combination therapy in patients. Our study aims to examine the effects of combination of erlotinib (E), an EGFR small-molecule inhibitor, and Alisertib (MLN8237, A), an inhibitor of Aurora A kinase, in lung cancer models. Treatment of two human NSCLC cell lines, A549 and H322M, with E+A showed synergistic activity in vitro. These cell lines were chosen for the xenografts studies in CB17 SCID mice. After tumors reached a volume of 100 mm3, mice were treated with E+A for three weeks. The xenografts were sensitive to growth inhibition by E (10 mg/kg/day) and A (20 mg/kg/twice daily) alone and the combination had an additive effect. In order to understand early cell signaling events in tumors treated with these agents we repeated the xenograft experiment by treating mice with a one time dosing of each drug alone and in combination. Three tumors per group were collected at 0 , 6 , 24 , or 48 hours after treatment and a panel of signaling molecules downstream of the EGFR and Aurora A kinase pathways were examined by Western blot. These included EGFR/pEGFR; HER2; HER3; MEK/pMEK; ERK/pERK; Src/pSrc and Akt/pAkt among others. Our results suggest that combined inhibition of both pathways in lung cancer cell lines and xenografts can lead to a reduced proliferative index as indicated by quantitative changes in these proteins and that this combination might be effective as a treatment strategy. A clinical trial with this combination is in progress at our center.

Citation Format: Tetyana Bagnyukova, Xiaofan Pan, Jiping Zhang, Brian Egleston, Igor Astsaturov, Erica Golemis, Hossein Borghaei. Effects of dual inhibition of EGFR and Aurora A kinase in a lung cancer model. [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 4242. doi:10.1158/1538-7445.AM2014-4242
Abstract 4249: The Pi3Kβ/δ inhibitor AZD8186 has potential to treat tumours in combination with key signalling pathway inhibitors
Authors: Barry Simon T. , Cronin Kathryn , Cumberbatch Marie , Ellston Rebecca , Foster Emily , Hancox Urs , Hanson Lyndsey , Harrington Liz , Lenaghan Carol , Symeonides Stefan , Trigwell Cath , Ward Lara
Published: May 2015

AZD8186 inhibits Pi3K isoforms Pi3Kβ and δ, with selectivity over Pi3Kα and γ. In solid tumours Pi3Kβ drives tumour growth when the tumour suppressor PTEN is deleted, mutated or downregulated. It also mediates signals from specific GPCR receptors. Pi3Kδ signals downstream of the B-cell receptor, creating potential for targeted treatment of haematological malignancies such as CLL, MCL & indolent NHL and possibly DLBCL. AZD8186 is differentiated from many other agents that target Pi3K family members as it isn't likely to impact glucose control. AZD8186 has single agent activity in a range of models, although maximal benefit is anticipated when used in combination. Loss of PTEN mediated control, and hence dependency on PI3Kβ occurs in many solid tumour types, and is altered in up to 40-50% of tumours in some individual disease states. Commonly PTEN dysregulation is associated with other activation of other signaling pathways. To explore this we have combined AZD8186 with a number of different agents. In HCC70 (triple negative breast cancer) and PC3 (prostate cancer) xenografts combination of AZD8186 (25mg/kg bid) with single dose docetaxel (15mg/kg) gives >90% tumour growth inhibition (TGI), compared to 40-50% with docetaxel alone. AZD8186 also combines with other targeted agents. In HCC70, AZD8186 (25mg/kg bid) combined with selumetinib (10mg/kg) gave 94% TGI compared to 66%, and 47% with each single agent. In combination with AZD2014 (mTORC1/2 inhibitor) (15mg/kg qd) in HCC70 xenografts AZD8186 (25mg/kg bid) gave regressions (-23%) compared to TGI of 87% and 77% for each agent alone. In 786-0 (renal cancer) xenografts AZD8186 (12.5mg/kg bid) and AZD2014 (15mg/kg qd) gave regression of -82% compared to TGI of 33% and regression of -39% with each agent alone. This data establishes the potential for AZD8186 to be used in combination with a number of different agents including the ability to customise dose and schedule to optimise both tolerability as well as anti-tumour effects. Further exploration of the combination opportunities for AZD8186 with other molecular targeted agents would inform on the potential for inhibitors of Pi3Kβ and δ to give benefit in different tumour types.

Citation Format: Simon T. Barry, Kathryn Cronin, Marie Cumberbatch, Rebecca Ellston, Emily Foster, Urs Hancox, Lyndsey Hanson, Liz Harrington, Carol Lenaghan, Stefan Symeonides, Cath Trigwell, Lara Ward. The Pi3Kβ/δ inhibitor AZD8186 has potential to treat tumours in combination with key signalling pathway inhibitors. [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 4249. doi:10.1158/1538-7445.AM2014-4249
Abstract 4252: Regulation of the oncogene ZNF217 by localization in breast cancer
Authors: Messana Matthew J. , Yang Chao , Littlepage Laurie E.
Published: May 2015

Poor prognosis in breast cancer patients occurs when malignant tumors adapt to environmental insults, become resistant to chemotherapy, evade immune surveillance and metastasize to other tissues. Tumors are thought to arise from individual cells with multiple mutations, including amplification of genomic regions that provide a growth advantage. A region on human chromosome 20 called 20q13 is increased in ∼25% of early stage human breast cancers and correlates with poor prognosis in patients. We have studied a novel oncogene ZNF217 within this region. ZNF217 is key in promoting breast cancer: it is not only a prognostic indicator of breast cancer progression in patients who have the worst prognosis but also is itself a drug target and/or a marker of patient response to therapy. We find that ZNF217 protein is expressed most strongly in a small subset of cells within normal mammary epithelium and localizes predominantly in the nucleus of mammary epithelial cells. In contrast, the localization of ZNF217 is heterogeneous in breast tumors, with localization in both the nucleus and cytoplasm. Moreover, a truncated form of the protein localizes exclusively in the cytoplasm. We hypothesize that ZNF217 cytoplasmic localization affects ZNF217 function during cancer progression and can be used to predict poor prognosis in breast cancer patients. Thus, our first aim is to identify the regions of ZNF217 that are required for ZNF217 localization in the nucleus and cytoplasm. Second, we will utilize integrated biological approaches to determine the consequences of mislocalization of ZNF217 on cancer progression. We will determine which regions of ZNF217 are required for oncogenesis by overexpressing ZNF217 truncations in mammary epithelial cells and using these cells in a variety of assays that model steps in cancer progression_assays that have previously shown that ZNF217 promotes cancer progression. If our data shows that the cytoplasmic ZNF217 is implicated in oncogenesis, further studies will be required to identify the mechanisms by which ZNF217 is truncated in cells. Hopefully, this will aid in the development of innovative cancer diagnostics and therapeutics for breast cancer and other cancers.

Citation Format: Matthew J. Messana, Chao Yang, Laurie E. Littlepage. Regulation of the oncogene ZNF217 by localization in 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 4252. doi:10.1158/1538-7445.AM2014-4252
Abstract 4254: ESE1 binds to beta catenin and modulates its expression
Authors: Yang Xu-yu , Kim Kui Jin , Shao Hong Jun , Lee Seong-Ho
Published: May 2015

ESE-1 is an epithelium-specific transcription factor that functions in the differentiation of intestinal epithelial cells. ESE-1's structure predicts its interaction with a number of other proteins with various cellular functions. Previously we have shown that tolfenamic acid (TA) could induce nuclear translocation of ESE1 in colorectal cancer cells, and plays a critical role in the activation of apoptosis. Here we show that β-catenin, an important protein regulating cell-cell adhesion and gene transcription, binds and colocalizes with ESE-1 in cell cytoplasm. The binding sites were mapped to carboxy terminus ESE-1, which includes the ETS domain, and N-terminus of β-catenin, which contains several important phosphorylation sites. C-terminus of ESE1 also binds to α-catenin, and ESE1/ β-catenin interaction was abrogated with suppression of either β-catenin or α-catenin in HEK293 cells by si RNA treatment; moreover, suppression of both α-catenin and β-catenin further impaired ESE1/ β-catenin binding. Furthermore, knockdown of endogenous ESE1 significantly decreased β-catenin level in HCT116 cells. Current data support a role of ESE1 in the regulation of β-catenin function in colon cancer.

Citation Format: Xu-yu Yang, Kui Jin Kim, Hong Jun Shao, Seong-Ho Lee. ESE1 binds to beta catenin and modulates its expression. [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 4254. doi:10.1158/1538-7445.AM2014-4254