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reproductive tract rho rho GTPase-activating protein transcription factor transcriptome sequencing tumor tumorigenesis unpublished works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.

抽象的最近对人类癌症基因突变的分析意外地发现了编码 p190A 的 ARHGAP35 RhoGAP（p190A）作为高度突变基因，即前30名泛癌基因。突变尤其丰富腺癌，包括肾细胞癌。 ARHGAP35 的突变谱与作为肿瘤抑制剂的作用。此外，ARHGAP35 基因座位于基因组中经常出现的区域。因癌症而消失。然而，生物信息学数据不足以确定基因的功能后果突变。因此，该提案的范围是定义与 p190A 丢失相关的致癌能力表达以及人类癌症中自然发生的 ARHGAP35 突变。我们最近获得的结果表明 p190A 及其旁系同源物 p190B - 统称为 p190 在此 - 介导上皮细胞增殖的接触抑制 (CIP)。我们还进行了全基因组 RNA-seq 分析，并确定 p190 调节受 YAP 癌蛋白。 YAP 是 TEAD 家族转录因子的转录共激活因子。 YAP的活动受 Hippo 途径控制，该途径广泛参与 CIP。在目标 1 中，我们将定义 p190 影响 Hippo 信号传导和 CIP 的机制。为此，我们将 (i) 测试 Rho 蛋白是否需要和/或足以促进 p190 下游的 CIP； (ii) 确定是否 p190 通过规范和/或非规范 Hippo 途径发出信号； (iii) 确定 p190 是否影响 YAP- TEAD介导的基因表达； (iv) 确定 p190A 和 p190B 之间是否存在明显冗余上皮细胞是环境依赖性的。在目标 2 中，我们将阐明 p190 在粘附连接 (AJ) 上的功能，该功能在通过 Hippo 途径促进 CIP。为此，我们将 (i) 测试 p190 激活 Hippo 的作用新生 AJ 形成时的途径； (ii) 确定 E-钙粘蛋白恢复 Hippo 是否需要 p190 癌细胞中的信号传导； (iii) 定义 p190 在失巢凋亡中的作用，这是一种肿瘤抑制机制由 AJ 和 Hippo 通路调节； (iv) 阐明 p190 和 p120 之间是否存在相互作用- Hippo 信号传导和 CIP 需要连环蛋白。在目标 3 中，我们将与布罗德研究所的 Gad Getz 博士合作，分析人类肿瘤 ARHGAP35 突变导致 (i) 杂合性丧失； (ii) YAP调节基因的表达； (iii) 换算和独家数据。接下来，我们将确定自然发生的 ARHGAP35 突变对 p190A 的影响 RhoGAP 活性，以及 Hippo 信号传导和 CIP。最后，我们将在体外和体内测试是否表达外源性 p190A 会减弱人类癌细胞系的致癌能力，而没有或水平非常低内源性 p190A，并且这种效应是否依赖于 GAP 活性和/或 Hippo 信号传导。这些努力对于阐明靶向 Rho 信号传导在未来个性化肿瘤治疗中的效用至关重要。

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