p190 RhoGAP signaling in epithelial oncogenesis

p190 RhoGAP 信号在上皮肿瘤发生中的作用

基本信息

批准号：
10080030
负责人：
STEEN HENNING HANSEN
金额：
$ 40.49万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-02-03 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10080030
关键词：
Adenocarcinoma Adherens Junction Anoikis Area Attenuated Bioinformatics Cadherins Cancer cell line Carcinoma Catalogs Cell Communication Cell-Cell Adhesion Cells Censuses Collaborations Contact Inhibition Data E-Cadherin Endometrial Carcinoma Epithelial Epithelial Cells Family Future GRLF1 gene Gastrointestinal tract structure Gene Expression Gene Mutation Genes Genetic Transcription Genome Genomic Segment Goals Human Hydrolysis In Vitro Inhibition of Cell Proliferation Institutes Link Literature Loss of Heterozygosity Lung MDCK cell Malignant Epithelial Cell Malignant Neoplasms Mediating Mutate Mutation Oncogenes Oncogenic Oncology Oncoproteins Organ Pathway interactions Play Proteins Publishing Renal Cell Carcinoma Role Sampling Signal Pathway Signal Transduction Squamous cell carcinoma Testing Tissues Transcription Coactivator Tumor Suppressor Proteins Uncertainty Validation Work cancer genome cell type driver mutation genome-wide improved in vitro testing in vivo insight mRNA sequencing novel paralogous gene personalized cancer therapy precision oncology reproductive tract rho rho GTPase-activating protein transcription factor transcriptome sequencing tumor tumorigenesis unpublished works

项目摘要

ABSTRACT Recent analyses of gene mutations in human cancer unexpectedly identified ARHGAP35 encoding p190A RhoGAP (p190A) as a highly mutated gene, i.e. top 30 pan-cancer. Mutations were particularly abundant in adenocarcinomas, including renal cell carcinoma. The mutation spectrum for ARHGAP35 is consistent with a role as tumor suppressor. Moreover the ARHGAP35 locus is located in a region of the genome that frequently is lost in cancer. However, bioinformatics data stop short of establishing the functional consequences of gene mutations. The scope of this proposal is therefore to define oncogenic capacities associated with loss of p190A expression and with naturally occurring ARHGAP35 mutations in human cancer. We have recently obtained results demonstrating that p190A and its paralog p190B - collectively termed p190 here - mediate contact inhibition of proliferation (CIP) in epithelial cells. We moreover conducted a genome wide RNA-seq analysis, and determined that p190 modulates expression of genes regulated by the YAP oncoprotein. YAP is a transcriptional co-activator of TEAD family transcription factors. The activity of YAP is controlled by the Hippo pathway, which is widely implicated in CIP. In Aim 1 we will define mechanisms whereby p190 impacts Hippo signaling and CIP. To this end, we will (i) test whether Rho proteins are required and/or sufficient to promote CIP downstream of p190; (ii) determine if p190 signals through the canonical and/or non-canonical Hippo pathways; (iii) establish if p190 impacts YAP- TEAD-mediated gene expression; and (iv) determine if apparent redundancy between p190A and p190B in epithelial cells is context-dependent. In Aim 2 we will elucidate the function of p190 at adherens junctions (AJs), which play essential roles in promoting CIP through the Hippo pathway. In this aim, we will (i) test a role for p190 to activate the Hippo pathway upon formation of nascent AJs; (ii) establish whether p190 is required for E-cadherin to restore Hippo signaling in carcinoma cells; (iii) define a role for p190 in anoikis, a tumor suppressor mechanism that is modulated by both AJs and the Hippo pathway; and (iv) elucidate if an interaction between p190 and p120- catenin is required for Hippo signaling and CIP. In Aim 3 we will in collaboration with Dr. Gad Getz of the Broad Institute, analyze human tumors with ARHGAP35 mutation for (i) loss-of-heterozygosity; (ii) expression of YAP regulated genes; and (iii) co-mutation and exclusivity data. Next, we will determine the effects of naturally occurring ARHGAP35 mutations on p190A RhoGAP activity, as well as Hippo signaling and CIP. Finally, we will test in vitro and in vivo if expression of exogenous p190A attenuates oncogenic capacities of human cancer cell lines with no or very low levels of endogenous p190A, and if such effects are dependent GAP activity and/or on Hippo signaling. These efforts are essential to elucidate the utility of targeting Rho signaling in future personalized oncology therapy.

抽象的对人类癌基因突变的最新分析意外鉴定了编码P190A的ARHGAP35 Rhogap（P190A）作为高度突变的基因，即前30个泛伴侣。突变特别丰富腺癌，包括肾细胞癌。 ARHGAP35的突变光谱与A一致作为肿瘤抑制剂的作用。此外，ARHGAP35基因座位于基因组的区域中在癌症中丢失。但是，生物信息学数据停止无法建立基因的功能后果突变。因此，该提案的范围是定义与损失P190A相关的致癌能力人类癌症中的表达和天然存在的ArhGAP35突变。我们最近获得了结果，证明了P190A及其旁系同源物P190B-统称 P190这里 - 介导上皮细胞中增殖（CIP）的接触抑制。此外，我们进行了基因组广泛的RNA-seq分析，并确定p190调节了由 YAP癌蛋白。 YAP是TEAD家族转录因子的转录共激活因子。 YAP的活动由河马途径控制，该途径广泛与CIP有关。在AIM 1中，我们将定义机制，其中P190会影响河马信号传导和CIP。为此，我们将（i）测试是否需要Rho蛋白和/或足以促进P190下游的CIP；（ii）确定是否 P190通过规范和/或非典型河马途径信号；（iii）确定p190是否影响yap- Tead介导的基因表达；（iv）确定p190a和p190b之间的明显冗余上皮细胞与上下文有关。在AIM 2中，我们将在粘附连接处（AJS）阐明P190的功能，该功能在通过河马途径促进CIP。在此目标中，我们将（i）测试P190的角色以激活河马形成新生AJ的途径；（ii）确定e-钙粘蛋白是否需要P190恢复河马癌细胞中的信号传导； (iii) define a role for p190 in anoikis, a tumor suppressor mechanism that is 由AJ和河马途径调节；（iv）阐明p190和p120-之间的相互作用是否存在河马信号传导和CIP需要蛋白链蛋白。在AIM 3中，我们将与Broad Institute的Gad Getz博士合作，分析人类肿瘤（i）杂合性丧失的ARHGAP35突变；（ii）YAP调节基因的表达；（iii）共同致辞和排他性数据。接下来，我们将确定天然存在的ARHGAP35突变对P190A的影响 Rhogap活性以及河马信号传导和CIP。最后，我们将在体外和体内测试外源性P190A减弱了人类癌细胞系的致癌能力，没有或非常低内源性p190a，如果这些效应是依赖的间隙活性和/或河马信号传导。这些努力对于阐明未来个性化肿瘤疗法中靶向RHO信号传导的实用性至关重要。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Genome editing using FACS enrichment of nuclease-expressing cells and indel detection by amplicon analysis.

DOI：
10.1038/nprot.2016.165
发表时间：
2017-03
期刊：
Nature protocols
影响因子：
14.8
作者：
Lonowski LA;Narimatsu Y;Riaz A;Delay CE;Yang Z;Niola F;Duda K;Ober EA;Clausen H;Wandall HH;Hansen SH;Bennett EP;Frödin M
通讯作者：
Frödin M

p190 RhoGAP promotes contact inhibition in epithelial cells by repressing YAP activity.

DOI：
10.1083/jcb.201710058
发表时间：
2018-09-03
期刊：
The Journal of cell biology
影响因子：
0
作者：
Frank SR;Köllmann CP;Luong P;Galli GG;Zou L;Bernards A;Getz G;Calogero RA;Frödin M;Hansen SH
通讯作者：
Hansen SH

CRISPR/Cas9 Engineering of Adult Mouse Liver Demonstrates That the Dnajb1-Prkaca Gene Fusion Is Sufficient to Induce Tumors Resembling Fibrolamellar Hepatocellular Carcinoma.

DOI：
10.1053/j.gastro.2017.09.008
发表时间：
2017-12
期刊：
Gastroenterology
影响因子：
29.4
作者：
Engelholm LH;Riaz A;Serra D;Dagnæs-Hansen F;Johansen JV;Santoni-Rugiu E;Hansen SH;Niola F;Frödin M
通讯作者：
Frödin M