权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Determination of the role of CBP and p300 mediated Wnt signaling on colonic cells

确定 CBP 和 p300 介导的 Wnt 信号传导对结肠细胞的作用

基本信息

批准号：
7874159
负责人：
Michael Bordonaro
金额：
$ 18.32万
依托单位：
GEISINGER COMMONWEALTH SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-03-01 至 2014-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7874159
关键词：
Aftercare Apoptosis Binding Biological Biological Assay Biological Models Butyrates CREB-binding protein Cancer Cell Growth Cancer cell line Cell Culture Techniques Cell Differentiation process Cell Fraction Cell Line Cell Proliferation Cell physiology Cells Cessation of life Characteristics Chemopreventive Agent Colon Colon Carcinoma Colonic Neoplasms Colorectal Cancer Complex Dependence Development Disease EP300 gene Exhibits Flow Cytometry Future Gene Expression Genes Genetic Programming HCT116 Cells Heterogeneity Histone Deacetylase Inhibitor Human In Vitro Lead Link Maintenance Malignant Epithelial Cell Measures Mediating Medical Students Methodology Methods Modeling Morbidity - disease rate Neoplasm Metastasis Neoplasms Normal Cell Nuclear Pathway interactions Pattern Phenotype Play Population Prevention Proteins Relative (related person)Research Research Proposals Role Science Signal Pathway Signal Transduction Sorting - Cell Movement Staging Students Testing Therapeutic Training Transcription Coactivator Transfection Universities Variant Vorinostat adenoma beta catenin cancer cell cancer initiation cell type clinically significant cofactor design graduate student human CREBBP protein in vivo inhibitor/antagonist lymph nodes medical schools molecular oncology mortality neoplastic prevent programs public health relevance research study response small molecule tumor tumor progression tumorigenesis tumorigenic

项目摘要

DESCRIPTION (provided by applicant): The Wnt signaling pathway, mediated through active beta-catenin, is responsible for initiating the majority of cases of human colorectal cancer (CRC), and we have previously shown that hyper-activation of this pathway by histone deacetylase inhibitors (HDACis), such as butyrate, can induce the death of CRC cells. An important cellular switch that mediates the effects of Wnt signaling activation is variation in the association between beta-catenin and the transcriptional coactivators CREB binding protein (CBP) and p300. Association of CBP with beta-catenin is thought to activate a set of genes linked to cell proliferation, while the p300-mediated Wnt genetic program is believed to promote cell differentiation. Small molecule agents have been discovered that modulate CBP/p300 Wnt transcriptional programs by altering the association of CBP and p300 to beta-catenin. ICG-001 and ICG-427 inhibit CBP and p300 mediated Wnt activity, respectively, while IQ-1 prevents the shift from CBP-mediated to a p300-mediated Wnt activity. Aim 1 of this proposal is designed to determine the role of CBP and p300 mediated Wnt signaling in the response of CRC cells to HDACis. Cells will be cotreated with HDACis and ICG-001, ICG-427, or IQ-1 and the levels of Wnt activity, apoptosis, proliferation, differentiation, and CBP- or p300-beta-catenin binding measured. Aim 2 of this proposal is to determine the role of CBP/p300-mediated Wnt activity in the maintenance of high- and low-Wnt fractions in single CRC cell populations, which may mirror similar heterogeneity observed in human tumors and which may be of clinical significance. Aim 3 will compare the effects of CBP and p300 mediated Wnt activity on CRC initiation and progression, utilizing CRC cell line model systems of initiation and progression: the normal colon cell lines CCD-841CoN, the adenoma line LT97, the primary colon carcinoma cell line SW480, and the lymph node metastasis cell line SW620. Cells will be treated with HDACis and the small molecule agents, and assayed as described above. We will also attempt to use changes in CBP and p300 mediated Wnt signaling to shift colonic cells between cell type, modifying CBP and p300 mediated gene expression in the LT97 adenoma line to shift the adenoma phenotype to more characteristic of the CCD- 841CoN normal cells, or the SW480 carcinoma cells. We will utilize microarray analyses to determine the patterns of gene expression responsible for these CBP or p300 mediated changes in colonic neoplastic phenotype. The findings generated from this study will lead to future, more in-depth projects to further dissect the action of CBP/p300 Wnt-mediated transcriptional programs in colonic neoplasia, with an emphasis on methods to modulate these genetic programs for chemopreventive effect. PUBLIC HEALTH RELEVANCE: The Wnt signaling pathway is responsible for initiating the majority of cases of human colorectal cancer, and we have previously shown that hyper-activation of this pathway by histone deacetylase inhibitors (HDACis) can induce the death of colorectal cancer cells. Wnt signaling mediated by the proteins CBP or p300 differentially influence decisions of cancer cell growth, differentiation, or death, and small molecule agents exist that can specifically block CBP and p300 mediated Wnt signaling. Through the use of these small molecule inhibitors, we will determine how CBP and p300 influence colonic tumorigenesis; the findings of this study can be utilized to devise methodologies, targeting CBP and p300 mediated Wnt activity, to prevent the development and progression of human colorectal cancer, decreasing the morbidity and mortality associated with this disease.

描述（由申请人提供）：通过活性β-连环蛋白介导的Wnt信号通路是引发大多数人类结直肠癌（CRC）病例的原因，我们之前已经证明，组蛋白脱乙酰酶抑制剂（HDACis）过度激活该通路丁酸盐等可以诱导CRC细胞死亡。介导Wnt信号传导激活作用的一个重要细胞开关是β-连环蛋白与转录共激活因子CREB结合蛋白（CBP）和p300之间的关联变化。CBP与β-连环蛋白的结合被认为激活了一组与细胞增殖相关的基因，而p300介导的Wnt遗传程序被认为促进了细胞分化。已经发现小分子试剂通过改变CBP和p300与β-连环蛋白的结合来调节CBP/p300 Wnt转录程序。ICG-001和ICG-427分别抑制CBP和p300介导的Wnt活性，而IQ-1阻止从CBP介导的Wnt活性转变为p300介导的Wnt活性。本提案的目的1旨在确定CBP和p300介导的Wnt信号传导在CRC细胞对HDACis的应答中的作用。将用HDACis和ICG-001、ICG-427或IQ-1共处理细胞，并测量Wnt活性、凋亡、增殖、分化和CBP-或p300-β-连环蛋白结合的水平。该提案的目的2是确定CBP/p300介导的Wnt活性在单个CRC细胞群中维持高和低Wnt组分中的作用，这可能反映了在人类肿瘤中观察到的相似异质性，并且可能具有临床意义。目的3利用大肠癌细胞系模型系统：正常结肠细胞系CCD-841 CoN、腺瘤细胞系LT 97、原发性结肠癌细胞系SW 480和淋巴结转移细胞系SW 620，比较CBP和p300介导的Wnt活性对大肠癌发生和进展的影响。细胞将用HDACis和小分子试剂处理，并如上所述进行测定。我们还将尝试使用CBP和p300介导的Wnt信号传导的变化来在细胞类型之间转移结肠细胞，修改LT 97腺瘤系中CBP和p300介导的基因表达，以将腺瘤表型转移到更具CCD-841 CoN正常细胞或SW 480癌细胞特征的细胞。我们将利用微阵列分析来确定负责这些CBP或p300介导的结肠肿瘤表型变化的基因表达模式。这项研究的结果将导致未来更深入的项目，以进一步剖析CBP/p300 Wnt介导的转录程序在结肠肿瘤中的作用，重点是调节这些遗传程序以实现化学预防效果的方法。公共卫生相关性：Wnt信号通路是引发大多数人结直肠癌病例的原因，我们以前已经证明，组蛋白脱乙酰酶抑制剂（HDACis）过度激活该通路可以诱导结直肠癌细胞死亡。由蛋白CBP或p300介导的Wnt信号传导差异性地影响癌细胞生长、分化或死亡的决定，并且存在可以特异性阻断CBP和p300介导的Wnt信号传导的小分子试剂。通过使用这些小分子抑制剂，我们将确定CBP和p300如何影响结肠肿瘤发生;本研究的结果可用于设计靶向CBP和p300介导的Wnt活性的方法，以预防人类结直肠癌的发展和进展，降低与这种疾病相关的发病率和死亡率。