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

A long noncoding RNA antagonizes the PBAF complex to promote ccRCC progression

长非编码 RNA 拮抗 PBAF 复合物，促进 ccRCC 进展

基本信息

批准号：
9982035
负责人：
Rebekah Brooks
金额：
$ 3.26万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9982035
关键词：
Address Affect Binding Biological Assay Bromodomain Carcinoma Cell Cycle Regulation Cell Line Cells Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Code Complex Conventional (Clear Cell) Renal Cell Carcinoma Data Ectopic Expression Feedback Future Genes Genetic Transcription Genomics Glutamine Glycolysis Grant Growth Hypoxia Inducible Factor Individual Kidney Neoplasms Knock-out Lead Lipids Malignant Epithelial Cell Mediating Messenger RNA Metabolic Metabolism Mutate Mutation Normal tissue morphology Patients Proteasome Inhibitor Proteins Proteome Proteomics RNA RNA Binding RNA-Binding Proteins Renal carcinoma Repression Role SMARCA4 gene Sucrose System TRIM25 gene Translations Tumor Burden Tumor Suppressor Proteins Tumor Volume Ubiquitination Untranslated RNA VHL protein Xenograft procedure circadian differential expression experimental study in vivo knock-down metabolomics molecular clock mutant novel therapeutic intervention overexpression polybromo promoter protein expression scaffold tumor tumor growth tumor progression tumorigenesis ubiquitin-protein ligase

项目摘要

Project Summary: We discovered that a completely uncharacterized long noncoding RNA (lncRNA) ADIRF- AS1 binds to the tumor suppressor Polybromo- and BRG1-associated factors-containing (PBAF) chromatin modifying complex, including the three unique protein components of the PBAF complex: ARID2, BRD7, and PBRM1. The PBAF complex bromodomain containing protein PBRM1 is mutated in approximately 40% of clear cell renal carcinoma (ccRCC), where loss of PBRM1 increases proliferation and overall tumor burden. Publicly available patient data revealed that ADIRF-AS1 is more highly expressed in ccRCC tumors compared to normal tissues. We overexpressed ADIRF-AS1 in a panel of ccRCC cell lines and found that ADIRF-AS1 overexpression significantly promoted proliferation only in ccRCC cell lines with wildtype expression of PBAF protein components. Importantly, we found that overexpression of ADIRF-AS1 decreased protein expression of PBAF protein components. Next, we sought to identify how ADIRF-AS1 negatively regulates the PBAF complex. We performed a lncRNA pulldown assay followed by proteomics analysis and found that along with components of the PBAF complex, ADIRF-AS1 also associates with an E3 ubiquitin ligase TRIM25, which has been shown to target PBRM1 for proteasomal degradation. We validated that silencing TRIM25 increases PBRM1 expression and found that proteasome inhibitors rescued decreased expression of PBRM1 after ADIRF-AS1 overexpression. Therefore, I hypothesize that ADIRF-AS1 interacts with the PBAF complex to promote its degradation by acting as a scaffold for PBRM1 and TRIM25, promoting tumor formation and progression. To address this hypothesis, we propose two aims to: (1) Determine the role of ADIRF-AS1 expression in PBAF mediated metabolism and tumorigenesis, and (2) Characterize the mechanism by which ADIRF-AS1 negatively regulates the PBAF complex through TRIM25 to promote tumorigenesis.

项目概述：我们发现了一种完全未知的长链非编码RNA (lncRNA) ADIRF-