bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2381: The effect of ERK5 inhibition in clear cell renal cell carcinoma
Authors: Kanno Hidenori , Naito Sei , Ichiyanagi Osamu , Tsuchiya Norihiko 
Published: July 2017

Abstract
Introduction Clear-cell renal cell carcinoma (ccRCC) is the most prevalent kidney cancer and is characterized by von Hippel-Lindau tumor suppressor gene (VHL) mutation. VHL protein acts in a ubiquitin ligase complex, mediating the proteasomal degradation of ubiquitinized proteins, including hypoxia-inducible factor (HIF). Therefore, HIF accumulates in VHL-defective cell lines. A recent study revealed that extracellular signal-regulated kinase 5 (ERK5) is degraded through the ubiquitin-proteasome system, in a process mediated by VHL and HIF. Our objectives were to examine the effects of ERK5 in ccRCC and to investigate ERK5 as a potential therapeutic target. Methods ERK5 expression was investigated in surgically resected ccRCC specimens using immunohistochemistry. To confirm that ERK5 is degraded by the VHL pathway, ERK5 levels were examined by western blotting in Caki-1 VHL wildtype renal cell carcinoma (RCC) cells and in A498 and A704 VHL-mutant RCC cells with or without the MG132 proteasome inhibitor. Furthermore, VHL-mutant cell lines were examined, with or without siRNA and XMD8-92-mediated ERK5 inhibition, to investigate the role of ERK5 in RCC cells using flow cytometry, MTS assays, and western blotting . Results In surgical specimens, RCC cells showed higher levels of ERK5 expression than did normal cells. In addition, MG132 enhanced ERK5 expression in wildtype cell lines, but not in VHL-mutant cell lines. Therefore, ERK5 degradation involves the VHL pathway. Furthermore, ERK5 inhibition resulted in increase of the sub-G1 population as observed by flow cytometry and cleaved RARP expression as seen by western blotting, indicating an increase in apoptotic cells. ERK5 inhibition also suppressed cell viability and downregulated p16 and Bcl-2 expression. Conclusion Our results show that ERK5 inhibition contributes to the downregulation of anti-apoptotic and cell cycle proteins and suggest that ERK5 is a promising therapeutic target for ccRCC.
Citation Format: Hidenori Kanno, Sei Naito, Osamu Ichiyanagi, Norihiko Tsuchiya. The effect of ERK5 inhibition in clear cell renal cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2381. doi:10.1158/1538-7445.AM2017-2381
Abstract 2382: RACK1 regulates PKA activity to drive tumorigenesis in colon cancer
Authors: Hayes Sheri L. , Dowling Catríona M. , Coffey John C. , Kiely Patrick A. 
Published: July 2017

Abstract
RACK1 (Receptor for Activated C Kinases) is a scaffolding protein with 7 WD repeats that interacts with the Insulin-like Growth Factor I receptor (IGF-IR), integrins, and other signalling proteins. RACK1 functions as a scaffolding protein and regulator of many key biological processes and it is highly expressed in most tissue. We recently reported that RACK1 is essential for IGF-I-mediated regulation of PP2A activity and AGAP2 activity at focal adhesions. Downstream of IGF-1R signaling in cancer cells, the scaffolding properties of RACK1 are altered providing distinct migratory advantages to the cell and suggesting that RACK1 is an important regulator of IGF-I signalling in cancer progression. It is important to further characterise the role of RACK1 in cancer and to delineate the molecular mechanisms by which it regulates key signalling pathways. We have identified Protein Kinase A (PKA) as RACK1 interacting protein. PKA is a complex, multicomponent enzyme which is a fundamental protein that functions in cell survival, proliferation, and cytoskeletal remodelling among others. Our hypothesis is that RACK1 regulates the PKA axis in cancer cells leading to Akt and MAP Kinase activation downstream of IGF-1R signalling. We have demonstrated that RACK1 is required to mediate PKA activity in colon cancer cells. Although we show that PKA expression is not altered in colon cancer tissue, we believe that PKA activity is altered significantly in the disease state. Using PKA inhibitors, we have shown that PKA regulates IGF-I-mediated MAP Kinase activation in colon cancer cell lines. In conclusion, this research project will help elucidate the role of PKA in colon cancer. Inhibiting or promoting specific protein interactions with RACK1 will provide very novel therapeutic opportunities and anti-cancer drug targets.
Citation Format: Sheri L. Hayes, Catríona M. Dowling, John C. Coffey, Patrick A. Kiely. RACK1 regulates PKA activity to drive tumorigenesis in colon cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2382. doi:10.1158/1538-7445.AM2017-2382
Abstract 2384: Aberrant expression of PDZ binding kinase modulates human pancreatic cancer cell invasion
Authors: Hinzman Charles , Trevino Jose , Banerjee Partha P. 
Published: July 2017

Abstract
Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer-related deaths and projected to be the second leading cause of cancer deaths by 2030. Incidence to mortality ratio for PDAC is almost one suggesting its aggressive phenotype. The poor survival rate of PDAC patients is due to metastasis to distant organs, and therefore, recent focus has been aimed towards the identification of molecular targets that drives metastatic disease. We are investigating an under-studied kinase, PDZ-domain binding kinase (PBK) as a target for pancreatic cancer (PanCa) therapy. PBK is a dual-specificity serine/threonine kinase involved in mitosis and invasive behavior of cancer cells. It is expressed physiologically in testis, but aberrant expression results in an aggressive phenotype in a variety of cancers. Our results demonstrate that PBK expression varies across a panel of human PanCa cell lines, and directly correlates with their invasive ability. Similarly, a panel of patient-derived primary PanCa shows higher PBK levels associated with aggressive behavior and invasion. To assure specificity of PBK to this invasive phenotype, we measured the effect of PBK overexpression or knockdown on the PanCa lines’ invasive ability. Overexpression of PBK in low PBK expressing cells increased PanCa invasion ~10-fold, while PBK knockdown reduced invasive ability ~5-fold. Immunohistochemical analyses of resected human PanCa specimens revealed that PBK is present in the majority of human PanCa tissue samples tested but not in the adjacent normal pancreatic tissue. Interestingly, aberrant PBK localization was found in even early stages of PanCa development and was sustained throughout its progression. In search of molecular effectors of invasion regulated by the PBK, we examined various genes/proteins that are involved in cancer cell invasion and metastasis. RelB a well-known metastatic inducer in various cancers was found to be a major payer in PanCa and regulated by PBK. These data indicate a novel molecular pathway of PBK where its presence plays a role in invasion of human PanCa cells. Therefore, the role of PBK in promoting cancer cell invasion, combined with its general lack of expression in normal cells, nominates PBK as a potentially important therapeutic target for pancreatic cancer.
Citation Format: Charles Hinzman, Jose Trevino, Partha P. Banerjee. Aberrant expression of PDZ binding kinase modulates human pancreatic cancer cell invasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2384. doi:10.1158/1538-7445.AM2017-2384
Abstract 2386: HDAC6 regulates ERK1’s kinase activity via deacetylation
Authors: Wu Jheng-Yu , Zhang Xiaohong 
Published: July 2017

Abstract
Extracellular signal-regulated kinases 1/2 (ERK1/2) are important kinases regulating cell proliferation and cell migration, and have been established as therapeutic targets for cancer treatment. Previously, we found that ERK1 phosphorylates histone deacetylase 6 (HDAC6), a class IIb HDAC, to regulate its enzymatic activity. HDAC6 plays an important role in several types of cancers. HDAC6 governs cancer cells’ migration by cooperating with cytoskeleton proteins and their associated proteins, and HDAC6 is arising as a determinant for cisplatin-resistance in non-small cell lung cancer (NSCLC) and ovarian cancer. Here, we have shown that HDAC6 manipulates ERK1’s kinase activity via deacetylation. We demonstrated that both ERK1 and ERK2 interact with HDAC6 physically via GST pull-down assays. To examine whether ERK1/2 are acetylated, we transfected GST-ERK1 or GST-ERK2 into HEK293T cells, and treated the cells with a pan-HDAC inhibitor, Trichostatin A. We showed that the level of acetylated GST-ERK1/2 increased in a dose- and time-dependent manner. Furthermore, when GST-ERK1 or GST-ERK2-expressed cells were treated with an HDAC6-specific inhibitor, ACY-1215, the level of acetylated GST-ERK1/2 increased. These results suggest that HDAC6 is a deacetylase for ERK1/2. In addition, we determined that acetyltransferases CBP and p300 acetylate ERK1/2. We found a novel acetylation site located in ERK1 N-terminus by mass-spectrometry analysis. We then mutated this lysine (K) site with either Glutamic acid (Q) or Arginine (R) to mimic the acetylation or deacetylation status, and performed kinase assays using ELK1 as a substrate. The acetylation mimicking mutant exhibits a decreased kinase activity toward ELK1, while the deacetylation mimicking mutant exhibits a similar level of kinase activity as the wild-type ERK1 does, suggesting that acetylation/deacetylation alters ERK1 enzymatic activities. Therefore, HDAC6 may regulate ERK1’s kinase activity via deacetylation of this site. Furthermore, we have shown that HDAC6 is overexpressed in lung cancer tissue samples and plays an important role in chemo-sensitivity in lung cancer. We predict that inhibition of the HDAC6-ERK1 pathway may provide clinical benefit for lung cancer patients.
Citation Format: Jheng-Yu Wu, Xiaohong Zhang. HDAC6 regulates ERK1’s kinase activity via deacetylation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2386. doi:10.1158/1538-7445.AM2017-2386
Abstract 2389: LINE-1 tumor hypomethylation is associated with shorter recurrence-free survival in localized clear-cell renal cell carcinoma
Authors: Malouf Gabriel G. , Mouawad Roger , Allanic Frederick , Compérat Eva , Roupret Morgan , Khayat David , Spano Jean-Philippe 
Published: July 2017

Abstract
Background: Cancer cells are characterized by alterations of DNA methylation patterns involving global DNA hypomethylation and site-specific CpG island promoter hypermethylation. Global DNA hypomethylation is thought to play a role in genetic instability and tumor aggressiveness. Long interspersed nucleotide element-1 (L1/LINE-1) repetitive elements represent 40% of the genome and their methylation is a good indicator of the global DNA methylation level. Although LINE-1 methylation has been previously shown to be associated with prognosis of patients with cancer, the value of LINE-1 methylation in predicting recurrence of patients with localized clear-cell renal cell carcinomas remains unknown.
Material and Methods: We quantified the LINE-1 methylation using bisulfite pyrosequencing in cohort of 200 patients with resected clear-cell renal cell carcinomas (AJCC stage I-III). LINE-1 methylation of adjacent normal kidney was also assessed in 128 cases. Threshold of tumor LINE-1 hypomethylation was defined as LINE-1 methylation in normal samples minus three standard deviation.
Results: Median methylation of tumor samples was 59.01% versus 61.61% for normal adjacent kidney samples (p&lt;10-5). LINE-1 methylation level of normal adjacent kidney was associated with tumor size (Spearman R=0.21, P=0.02) but not with age (p=0.24) or Leibovitch score (p=0.05). Tumor LINE-1 hypomethylation was observed in 20 samples (n=10%). Strikingly, those were enriched for tumors occurring in female (60% versus 27.7%) (p=0.005); of note, no association was found between LINE-1 tumor hypomethylation and tumor grade (p=0.31), stage (p=1), age (p=0.24) and Leibovitch score (p=0.82). In multivariate analysis, only Leibovitch score and LINE-1 hypomethylation were independently associated with poor recurrence-free survival.
Conclusion: LINE-1 hypomethylation is associated with shorter recurrence-free survival in patients with resected clear-cell renal cell carcinomas, suggesting the possibility of using it as predictive biomarker of recurrence.
Citation Format: Gabriel G. Malouf, Roger Mouawad, Frederick Allanic, Eva Compérat, Morgan Roupret, David Khayat, Jean-Philippe Spano. LINE-1 tumor hypomethylation is associated with shorter recurrence-free survival in localized clear-cell renal cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2389. doi:10.1158/1538-7445.AM2017-2389
Abstract 2390: Clinical validation of an epigenetic field of susceptibility to detect significant prostate cancer from 2 non-tumor biopsies
Authors: Yang Bing , McCormick Johnathon , Schultz Adam , Leverson Glen , Sonn Geoffrey , Magi-Galluzi Cristina , Klein Eric A. , Fumo Michael , Jarrard David F. 
Published: July 2017

Abstract
Background: An epigenetic field of cancer susceptibility exists for prostate cancer that gives rise to multifocal disease in the peripheral zone. In a previous studies (Neoplasia 2013, J Urol 2014) we identified using methylation arrays over 50 altered regions in normal prostate tissue of men with prostate cancer. In the current multicenter study, a validation was performed to determine the predictive strength of this approach.
Methods: We evaluated 2 archived, cancer negative prostate biopsy core tissue from 129 subjects from 4 urological centers. All negative cases (controls) underwent 2 or more repeat negative biopsies within 24 mo with a central review of all histopathology. Cancer cases were negative biopsies selected from patients who ultimately went prostate removal to confirm a final grade &gt;Gleason Score 7. Biopsies were analyzed using pyrosequencing for DNA methylation changes at multiple CpGs surrounding the genes EVX1, CAV1, PLA2G16, FGF1, SPAG4 and NCR2. Analyses used multiplex logistic regression modeling with backward elimination.
Results: Patients diagnosed with GS&gt;7 cancer (77) and the control group (52) were similarly matched except for PSA (7 vs 5.8; p&lt;0.03) and prostate size (46 vs 36; p&lt;0.02). We observed robust methylation differences for all genes (p&lt;0.05) (Table). A multiplex regression model for biomarker performance incorporating a gene combination of EVX1, NCR2, PLAG2 and SPAG4 discriminated non-tumor from tumor-associated tissues (AUC 0.744, p&lt;0.0001). A multiplex model incorporating 3 genes (EVX1, PLA2G16, NCR2) and clinical information (PSA, age) identified patients with GS&gt;7 prostate cancers (AUC 0.808, p=0.001).
Conclusions: A widespread epigenetic field defect can be used to detect GS&gt;7 prostate cancers in patients with histologically negative biopsies. This assay is sensitive in requiring only 2 biopsies and detects alterations in nontumor cells at distance from the cancer. This has the potential to decrease the need for repeat prostate biopsies, a procedure associated with cost and complications.
Support: UW Ignitor; Gen X foundation
Citation Format: Bing Yang, Johnathon McCormick, Adam Schultz, Glen Leverson, Geoffrey Sonn, Cristina Magi-Galluzi, Eric A. Klein, Michael Fumo, David F. Jarrard. Clinical validation of an epigenetic field of susceptibility to detect significant prostate cancer from 2 non-tumor biopsies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2390. doi:10.1158/1538-7445.AM2017-2390
Abstract 2392: A multiplex histone H3 PTM assay for epigenetic biomarker discovery in tissue biopsy and archived clinical samples
Authors: Jelinek Mary Anne , Ritland Melissa , Westergren AJ 
Published: July 2017

Abstract
Dysregulation of epigenetic mechanisms is known to play an important role in the development and progression of cancer. One such mechanism is manifested as altered levels of histone modifications involved in regulating gene transcription. N-terminal histone tails can have a variety of modifications, such as phosphorylation, methylation and acetylation at specific amino acid residues which are conserved throughout eukaryotes and function by altering chromatin structure and creating binding sites for chromatin readers, writers or erasers.
Numerous studies have reported aberrant global levels of several histone H3 and H4 post-translational modifications (PTMs) in a wide range of solid tumor types. These changes have been shown to be predictive of clinical outcome, raising the possibility that histone modifications have potential as epigenetic biomarkers. Clinical samples have immense potential for biomarker identification since they are often accompanied with valuable information pertaining to patient history, treatment courses and disease outcome. The preferred method for clinical sample preservation is formalin-fixation followed by paraffin embedding (FFPE). While extraction and downstream analysis of DNA and RNA from FFPE samples is now routine, proteomic studies of FFPE samples is hampered by extensive protein cross-linking generated by formalin fixation. Analysis of histone PTMs in patient archival samples is limited to low-throughput immunohistochemical staining, and is not an ideal approach for mining large sample cohorts for biomarker identification. Western blot or ELISA methods, which have large sample requirements, are simply not an option for FFPE samples.
We have developed a multiplex bead-based ELISA assay which enables simultaneous interrogation of thirteen histone H3 PTMs. The assay is performed in 96-well plates and is ideally suited for profiling histone modification levels in limiting or small samples. We will present the application of this technology in a variety of sample types including frozen tissue, formaldehyde-cross linked cells and tissues, and in small molecule inhibitor screens with as few as 2,000 cells per well. The ability to simultaneous detect up to thirteen histone H3 PTMs provides a unique feature that enables determination of for on-target and off-target effects within the same sample.
Citation Format: Mary Anne Jelinek, Melissa Ritland, AJ Westergren. A multiplex histone H3 PTM assay for epigenetic biomarker discovery in tissue biopsy and archived clinical samples [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2392. doi:10.1158/1538-7445.AM2017-2392
Abstract 2393: Decreased expression of 5-hydroxymethylcytosine is associated with grade of malignancy in lung cancer
Authors: Tominaga Yoshiaki , Ito Ken-ichi , Nakayama Jun 
Published: July 2017

Abstract
Purpose: The ten-eleven translocation (TET) proteins are critical regulators in epigenetic modification. It is well known that TET proteins catalyze the oxidation of 5-methylcytosine (5-mC) to generate 5-hydroxymethylcytosine (5-hmC), and 5-hmC levels reflect diverse functions of TET proteins. It has been reported that the expression of 5-hmC in cancer tissues is decreased compared with that in normal tissues in several organs. However, 5-hmC expression in lung cancer has not been fully analyzed. To examine whether or not 5-hmC was associated with phenotypes or progression of lung cancer, we evaluated 5-hmC expression in a variety of histological types of lung cancers.
Materials and methods: The expression of 5-hmC was analyzed by immunohistochemistry in 109 paired specimens of lung cancer and adjacent normal tissues obtained from the patients underwent operation at Shinshu University Hospital. Eighty-nine adenocarcinomas, 17 squamous cell carcinomas, two large cell carcinomas, and one small cell carcinoma were analyzed in the study.
Results: 5-hmC expression in female was significantly lower than that in male, and was lower in smokers than non-smokers. 5-hmC expression was positive in 34.1±22.0% of lung cancer tissues and 69.4±18.8% of normal tissues. The expression of 5-hmC was significantly decreased in the lung cancer tissues compared with that in the adjacent normal tissues (p&lt;0.001). With regard to the histological types, the positive ratios of 5-hmC expression were 36.0±21.7% in adenocarcinoma, 24.0±20.3% in squamous cell carcinoma, 8.0±7.3% in large cell carcinoma, and 13.0% in small cell carcinoma. Thus, the frequency of positive expression of 5-hmC was decreased in accordance with the increase of grade of malignancy of lung cancer, and a negative correlation between 5-hmC expression and the grade of malignancy of lung cancer was observed. On the other hand, no difference of the expression of 5-mC was observed between lung cancer tissues and normal tissues.
Conclusions: Our results indicate a possibility that 5-hmC may be related to carcinogenesis and progression or differentiation of lung cancer.
Citation Format: Yoshiaki Tominaga, Ken-ichi Ito, Jun Nakayama. Decreased expression of 5-hydroxymethylcytosine is associated with grade of malignancy in lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2393. doi:10.1158/1538-7445.AM2017-2393