bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 683: Toxicity of aerosolized azacytidine (Vidaza) in mice.
Authors: Qiu Xuan , Sellers Rani , Liang Yuanxin , Piperdi Bilal , Perez-Soler Roman , Zou Yiyu 
Published: August 2014

Abstract
Background: Azacytidine, a prototypical demethylation agent, has been approved to treat myelodysplastic syndrome. Commercial azacytidine (Vidaza) is delivered to patients by subcutaneous or intravenous administration. In a previous study, we found that aerosolized azacytidine can 1) inhibit human orthotopic lung cancers in a mouse xenograft model, 2) demethylate several promoters of tumor suppressor genes reported to be hypermethylated in human lung cancer, and 3) be better tolerated and more effective than systemic administration. These studies specifically addressed toxicity associated with aerosolized delivery of Vidaza to the lungs; specifically hematologic and pulmonary toxicity.
Methods: Vidaza was dissolved into sterile water for injection and aerosolized in a clinical standard aerosol system. Swiss mice male and female 6/group were restricted into a nose-only exposure system for receiving the aerosol. We treated the mice at 2 different doses, 7.5 mg/m2 daily x 7 days and 2.5 mg/m2 daily x 7 days. Blood, sternum (for bone marrow), and lung were collected at 3 and 6 weeks after the final aerosol treatment. The bone and lung were fixed in 10% neutral buffered formalin and routinely processed to paraffin blocks, sectioned, stained, and evaluated histologically. In a preliminary pharmacokinetic study, azacytidine concentrations in the blood and lung samples from mice receiving 2.5 mg/m2 of aerosolized azacytidine were determined by a MS-HPLC method previously described by others.
Results: Neither dose of aerosolized Vidaza resulted in bone marrow changes. A mild pneumonitis was present in lungs from mice treated with 7.5 mg/m2 at 3 weeks but was not evident at six weeks (reversible). No pulmonary toxicity was observed at the 2.5 mg/m2 dose. The preliminary pharmacokinetic study data indicate very low drug concentrations in the blood and much higher concentrations in the lungs.
Conclusion: Our study demonstrates that low dose aerosolized azacytidine can be safely administered to mice. Reversible pulmonary toxicity (pneumonitis) was observed at 7.5 mg/m2 at 3 weeks post treatment. The recommended starting dose of aerosolized azacytidine in human clinical phase I trial should be 2.5 mg/m2.
Supported by NIH grant 5R01CA154755 to Roman Perez-Soler and Yiyu Zou.
Citation Format: Xuan Qiu, Rani Sellers, Yuanxin Liang, Bilal Piperdi, Roman Perez-Soler, Yiyu Zou. Toxicity of aerosolized azacytidine (Vidaza) in mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 683. doi:10.1158/1538-7445.AM2013-683
Abstract 688: Promoter methylation of death receptor decoy genes in blood and risk of melanoma in melanoma-prone families .
Authors: Hyland Paula L. , Burke Laura S. , Pfeiffer Ruth M. , Tucker Margaret A. , Goldstein Alisa M. , Yang Xiaohong R. 
Published: August 2014

Abstract
Background: Cutaneous malignant melanoma (CMM) is an etiologically heterogeneous disease with genetic, environmental, and host characteristics and their interactions contributing to risk. Recently, gene-specific promoter methylation in blood has been associated with the risk of several cancers including breast, lung and pancreatic. We thus examined whether gene-specific promoter methylation was associated with risk of CMM in individuals from melanoma-prone families.
Methods: We measured the promoter methylation status of 15 genes (CDH1, COL1A2, DAPK, DDIT4L, HSPB6, LOX, MT1G, NPM2, p14, p16, PTEN, RASSF1, MAGEA3, TNFRSF10C and TNFRSF10D) previously shown to be aberrantly methylated in melanoma tumors using bisulfite pyrosequencing at 65 CpGs in peripheral blood mononuclear cells (PBMCs) from 236 individuals (114 melanoma cases and 122 unaffected individuals) in 64 melanoma-prone families. For functional validation we obtained gene expression data for 14 out of the 15 genes through independent expression profiling analysis. We also compared the values from each methylated promoter to mRNA expression. We used logistic regression to evaluate the association between CMM status and average promoter methylation controlling for age, sex and adjusting variance for familial correlation.
Results: In unaffected individuals, overall promoter methylation levels were low for most of these genes in blood. Only TNFRSF10C (P=0.009) and TNFRSF10D (P=0.010) showed the expected negative correlation between promoter methylation and gene expression. Therefore, we focused on these two genes in subsequent analyses. Compared to controls, CMM cases had significantly lower promoter methylation at TNFRSF10C (P= 0.002) and TNFRSF10D (P=0.045). This difference remained significant after further adjusting for CDKN2A, dysplastic nevi and pigmentation characteristics.
Significance: TNFRSF10C and TNFRSF10D encode separate decoy death receptors, whose overexpression in both normal and malignant cells can inhibit apoptosis. Our findings support a potential association between reduced promoter methylation of TNFRSF10C and TNFRSF10D in blood and risk of CMM in our melanoma-prone families.
Citation Format: Paula L. Hyland, Laura S. Burke, Ruth M. Pfeiffer, Margaret A. Tucker, Alisa M. Goldstein, Xiaohong R. Yang. Promoter methylation of death receptor decoy genes in blood and risk of melanoma in melanoma-prone families . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 688. doi:10.1158/1538-7445.AM2013-688
Abstract 716: 3-D imaging and illustration of nerve-lesion association in the mouse tongue of experimental oral cancer.
Authors: Hua Tzu-En , Liu Ko-Jiunn , Tang Shiue-Cheng 
Published: August 2014

Abstract
Introduction:
Tongue has a rich nerve supply and is in constant contact with a variety of neurotrophins in the saliva.
In tongue/oral cancer, patients suffer from pain that is often more severe than the symptom caused by other cancers, likely due to the stimulation of the nerve endings and/or compression and invasion of sensory nerves. Despite the noticeable neural component in tongue/oral cancer development, high-resolution microscopic observation of the tongue innervation in health and disease has been difficult. This is primarily due to the dispersed neural network in space that cannot be easily portrayed by the standard microtome-based 2-dimensional (2-D) microscopy. The artifact and distortion caused by microtome slicing as well as the challenge of aligning series of microtome slices in precision seriously limit our ability to examine tongue innervation. Method: To overcome the imaging limitation, we prepared transparent mouse tongue specimens by optical clearing (use of immersion solution to reduce random scattering as light travels across media; Fu & Tang, Gastroenterology,
139:p1100, 2010 and www.3d-histology.com) and combined vessel painting and
3-dimensional (3-D) neurohistology for joint visualization of the tongue tissue architectures. Cardiac perfusion of the fluorescent lectin was used to label the blood vessels. Neuronal markers including PGP9.5, tyrosine hydroxylase, vesicular acetylcholine transporter, and calcitonin gene-related peptide were used as the immunostaining targets of neural tissues. Results: We simultaneously revealed the microstructure, vasculature, and innervation of the normal tongue with μm-level resolution. Examples of 3-D features such as the neurovascular complex at the core of the filiform papilla and the taste bud innervation in the fungiform papilla were used to demonstrate the image quality. In the progression of experimental oral cancer induced by 4-Nitroquinoline-1-oxide, we observed remodeling of the tongue innervation with prominent peri- or intra-lesional nerve fibers, indicating intimate nerve-lesion interactions. Sympathetic, parasympathetic, and sensory nerves were all found associated with the lesion. Conclusion: This tongue innervation imaging method does not require tissue microtome sectioning and provides a useful tool for 3-D presentation and analysis of normal and diseased tongue in an integrated fashion.
Citation Format: Tzu-En Hua, Ko-Jiunn Liu, Shiue-Cheng Tang. 3-D imaging and illustration of nerve-lesion association in the mouse tongue of experimental oral cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 716. doi:10.1158/1538-7445.AM2013-716
Abstract 72: Targeting PI-3K/Akt signaling pathway by a PI-3K inhibitor BKM120 for radiosensitization in murine hepatocellular carcinoma.
Authors: Liu Wei-Lin , Gao Melissa , Cheng Ann-Lii , Cheng Jason Chia-Hsien 
Published: August 2014

Abstract
Purpose: Tumor control of hepatocellular carcinoma (HCC) by radiotherapy remains unsatisfactory and the sub-lethal effect associates with secondary spread. Phosphatidylinositol 3-kinase (PI-3K)/Akt pathway plays a critical role in promoting cancer cell proliferation and inhibiting cell death. Elevated PI-3K/Akt activity is associated with increased cellular resistance to irradiation. Our aim was to assess if the inhibition of PI-3K/Akt activity by a PI-3K inhibitor, BKM120, contributes to the increasing sensitivity of murine liver cancer cells to irradiation.
Methods and Materials: Murine HCC cell line (BNL) was used to evaluate the in vitro synergism of combining BKM120 with irradiation. Flow cytometry analyzed the cell cycle changes, whereas Western blot investigated the protein expressions after the combined treatment. Balb/c mice bearing ectopic BNL xenografts were treated with BKM120 and/or radiotherapy for the in vivo response.
Results: Clonogenic cell survival decreased dose-dependently either with irradiation or BKM120 treatment. Synergisms were achieved in cells treated together with greater or equal than 0.5μM BKM120 and 2 Gy radiation. BKM120 pretreatment significantly inhibited radiation-induced Akt phosphorylation. Western blots of active caspase-3 and cleaved PARP revealed that the combined BKM120 and radiation treatment strongly increased the expression of these apoptotic markers in response to irradiation compared with radiation alone. In ectopic xenografts, pretreatment with BKM120 significantly enhanced the tumor-suppressive effect by radiotherapy.
Conclusion: The apparent synergism between BKM120 and irradiation or effect of pre-treatment with BKM120 suggests an option to mitigate the activation of Akt by radiation, leading to increased cell apoptosis in murine liver cancer. These data suggest that the BKM120/radiation combination be a strategy worthy of further clinical trials.
Citation Format: Wei-Lin Liu, Melissa Gao, Ann-Lii Cheng, Jason Chia-Hsien Cheng. Targeting PI-3K/Akt signaling pathway by a PI-3K inhibitor BKM120 for radiosensitization in murine hepatocellular carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 72. doi:10.1158/1538-7445.AM2013-72
Abstract 724: New fluorescence-based cell assays for visualizing the Raf - Ras interaction and the Wnt pathway activity.
Authors: Apfel Johanna N. , Reischmann Patricia T. 
Published: August 2014

Abstract
Colorectal cancer (CRC) belongs to the most frequent cancer types (Hollande et al. 2010). As molecular mechanism for CRC, several mechanisms are presumed, including chromosome instability going along with the inactivation of the adenomatous polyposis coli (APC) gene, a key protein of Wnt signaling (Thorstensen et al. 2005). It is widely accepted that an abnormally activated Wnt pathway is a major cause for non-braked proliferation and for reduced cell differentiation. The second major cause of CRC is the deregulation of genes affecting the Ras pathway (Jeremy R. Jass, 2006).
Nevertheless, specific drugs targeting either the Wnt or the Ras pathways are rare. Therefore, we designed new, fluorescence-based reporter gene assays either for the Wnt or the Ras/Raf pathway as tools to simplify the screening for compounds modifying each pathway. Such cellular systems could be applicable for high-throughput-screening as well as Live-Cell-Imaging of compound effects, as shown in preliminary experiments. Therein, stably transfected HEK 293T cells bearing the new, Wnt-responsive reporter gene vector, were treated with LiCl, Wnt3a protein or acetylsalicylic acid in order to get a specific response. Cells showed an increase of the fluorescence level, which is measurable by a standard ELISA reader and could be visualized via fluorescence microscopy. In MDCK or MDCK-F3 cells bearing the Raf-responsive element, treated with EGF or a Raf inhibitor, the localization of the fluorescence signal changed. Moreover, treating the cells with different combination of the mentioned Wnt- or Ras-active compounds also showed a response, indicating a correlation between the two pathways. These results lead to the conclusion to combine both reporter gene vectors in order to detect the interplay of the pathways. Besides sensibilization of the assays and setting up the Live-Cell imaging experiments, this will be our further steps.
In summary, we designed several assays applicable in a screening assay for new, specific compounds targeting the two most important cancer-related pathways Wnt and Ras/Raf. Such an assay is more cost-effective than conventional assays, and also offers the possibility to characterize directly the biochemical mechanisms and interplays via Live-Cell-Imaging.
Citation Format: Johanna N. Apfel, Patricia T. Reischmann. New fluorescence-based cell assays for visualizing the Raf - Ras interaction and the Wnt pathway activity. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 724. doi:10.1158/1538-7445.AM2013-724
Abstract 736: Development of a novel circulating tumor cell technology for cancer predictive biomarker analysis.
Authors: Kang Zhigang , Dahut William , Gulley James , Yu Yunkai , Meltzer Paul S. , Cao Liang 
Published: August 2014

Abstract
Circulation tumor cells are critically important to understand tumor metastasis and may have a potential to noninvasively evaluate the cancer for clinical management. The isolation and characterization of CTC represent a major technological challenge. While the CellSeach system by J&J has shown that CTC enumeration can be an independent predictor of survival for breast, prostate and colon cancer, little molecular information can be obtained from the CTC to have an impact on patient management or clinical outcome. Most hormone dependent castration-resistant prostate cancer (CRPC) becomes refractory after one to three years of treatment, many through the amplification or mutation of the androgen receptor gene. As multiple targeted agents become available for this cancer and no tumor material can be obtained due to bone metastasis, there is an urgent need to be able to evaluate biomarkers of CRPC for clinical management of the patients. In order to assess biomarkers that may influence the management of CRPC, we worked on a novel antibody-independent CTC purification technology for isolation of Live CTC from CRPC.
Citation Format: Zhigang Kang, William Dahut, James Gulley, Yunkai Yu, Paul S. Meltzer, Liang Cao. Development of a novel circulating tumor cell technology for cancer predictive biomarker analysis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 736. doi:10.1158/1538-7445.AM2013-736
Abstract 742: Up-regulation of histone deacetylases in colon cancer.
Authors: Yang Hui , Salz Tal , Zajac-kaye Maria , Liao Daiqing , Huang Suming , Qiu Yi 
Published: August 2014

Abstract
The reversible acetylation of histones and non-histone proteins by histone acetyltransferases (HATs) and deacetylases (HDACs) play critical roles in transcriptional regulation in eukaryotic cells. Acetylation of histones is commonly associated with transcriptional activation of genes, and is thought to be responsible for the formation of a local “open chromatin” structure. In contrast, removal of acetyl groups by histone deacetylases (HDACs) frequently accompanies suppression of gene activity. Mammalian HDACs are classified into four classes (I, II, III and IV) based on the sequence homology to the yeast histone deacetylases Rpd3 (reduced potassium dependency), Hda1 (histone deacetylase1), and Sir2 (silent information regulator 2), respectively. Although the precise cellular functions of the different HDAC enzymes are still poorly understood, evidence suggests that different members of the HDAC family have distinct functions. It had been shown that overexpression of class I HDACs are more relevant to cancer development in many cancer types. In the absence of class I HDACs, cancer cells arrest either at the G1 phase of the cell cycle or the G2/M transition, resulting in the loss of mitotic cells, cell growth inhibition, and an increase in the percentage of apoptotic cells. Therefore, HDAC inhibitors had been used as a class of chemotherapeutic agents in cancer treatment. However, little is known about factors involved in regulation of class I expression in cancer cells. Our study focuses on the regulation of HDAC1 and 2 expressions in colon cancer cells. We found that compared with normal colon cell lines, both mRNA and protein level of HDAC1 and HDAC2 are higher in colon cancer cells, which suggests that HDAC1/2 is regulated at transcriptional level in colon cancer. HDAC1 and HDAC2 promoters were cloned and the promoter reporter activity was studied. Our result shows that HDAC1 and 2 promoters are highly active in colon cancer cells. Through mutation study on various putative transcription factors binding sites, we found that promoter activity is regulated by transcription factor Sp1/Sp3, which are also elevated in colon cancer cells lines and tissues. Chromatin immunoprecipitation assay also indicate that Sp1/Sp3 is recruited to HDAC1/HDAC2 promoter. Inhibition of Sp1/Sp3 through inhibitor or shRNA decreased HDAC1/2 expression and reduced cell proliferation. These results indicate that Sp1/Sp3 may be major regulators for HDAC1 and HDAC2 overexpression in colon cancer and this finding may shed light on future treatment of colon cancer.
Citation Format: Hui Yang, Tal Salz, Maria Zajac-kaye, Daiqing Liao, Suming Huang, Yi Qiu. Up-regulation of histone deacetylases in colon cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 742. doi:10.1158/1538-7445.AM2013-742
Abstract 745: Role of G-quadruplex structures in the human RET promoter region in the regulation of this gene.
Authors: Sun Daekyu , Shin Yoon-Joo 
Published: August 2014

Abstract
The medullary thyroid carcinoma (MTC) is a tumor type characterized by germline point mutations of the RET (REarranged during Transfection) proto-oncogene. Since these mutations are known to induce oncogenic activation of RET tyrosine kinase, various therapeutic strategies interfering with the oncogenic function of RET protein have been developed and tested for treating RET-associated cancers in several preclinical and clinical studies. Our previous study has demonstrated that the G-rich strand in the polypurine/polypyrimidine (pPu/pPy) region of the RET proximal promoter is capable of forming intramolecular G-quadruplex structures in vitro in the presence of K+, which can be further stabilized by a G-quadruplex interactive agents. Thus, the detailed mechanism by which G-quadruplex structures regulate RET transcription has been studied using mutation analysis and promoter reporter assays together with a detailed analysis of transcription factor binding to the RET promoter in vivo. To this end, we have explored the Flp-In cell system using homologous recombination, which allows integration of a single copy of a wild type or G-quadruplex-mutant RET promoter–luciferase construct into a specific genomic recombination locus (FRT site) in a host Flp-In the human embryonic kidney 293 (HEK293) cell line. Thus, differences in reporter expression levels from isolated isogenic cell clones can be ascribed more confidently to promoter-specific effects, rather than enhancer/silencer artifacts due to variations in integration sites. Using these isogenic cell lines, we are able to provide strong evidence for G-quadruplex-dependent transcriptional repression of the RET gene in vivo.
Citation Format: Daekyu Sun, Yoon-Joo Shin. Role of G-quadruplex structures in the human RET promoter region in the regulation of this gene. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 745. doi:10.1158/1538-7445.AM2013-745
Abstract 755: FK506 binding protein 51 (FKBP51) binds to p300 and acts as transcriptional co-regulator of ABCG2 gene expression in melanoma.
Authors: Romano Simona , Nappo Giovanna , Cesaro Elena , Candela Antonio , Romano Maria Fiammetta 
Published: August 2014

Abstract
Several lines of evidence support a role for FKBPs in chromatin modifications and cancer. Particularly for some of them, DNA association and regulation of gene expression has been demonstrated, mediated by histone- or transcription factor- chaperone activity. FKBP51 is an immunophilin that functions also as co chaperone of several proteins and receptors. We previously found this molecule has an essential role in melanoma resistance to the therapy and aggressiveness. Melanoma cells expressing stem cell markers (MCSC) produced high levels of FKBP51. In addition, the number of MCSC (identified by ABCG2 expression) was increased in FKBP51 over expressing melanoma cell lines. By converse, reduced expression of cancer stemness genes was measured in FKBP51 silenced melanoma. Since ABCG2+ cells also contained high levels of the general transcriptional co-activator, namely E1A binding protein p300, we hypotesized that FKBP51 might act as a p300 co chaperone with a role in transcriptional regulation of stemness markers. By co-IP assays, we show that FKBP51 interacts with p300 and partecipates in transcriptional complexes with Smad 2 and p300. By chromatin immunoprecipitation assays (ChIP) we demonstrate that FKBP51 binds to chromatin. We used different couples of primers to amplify different regions of the ABCG2 promoter, in FKBP51-immunoprecipitated chromatin and found enrichment of DNA fragments corresponding to a first sequence localized at 3450 bp (-3450) from the transcription start site (TSS), a second at -3219, and a third at -939. Differently, no enrichment in closest regions was found, suggesting the specificity of FKBP51 binding to ABCG2 promoter. In addition, we found p300 bound to the same regions of ABCG2 promoter, and such binding was impaired by FKBP51 downmodulation, suggesting the immunophilin regulatated the binding of p300 to DNA. In conclusion, our study supports the conclusion that FKBP51 partecipates in the transcriptional apparatus that controls expression of melanoma stemness genes, acting as co chaperone of p300.
Citation Format: Simona Romano, Giovanna Nappo, Elena Cesaro, Antonio Candela, Maria Fiammetta Romano. FK506 binding protein 51 (FKBP51) binds to p300 and acts as transcriptional co-regulator of ABCG2 gene expression in melanoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 755. doi:10.1158/1538-7445.AM2013-755