Directional Motility and ERM Scaffolding in Pathfinder Pancreatic Carcinoma Cells

Pathfinder 胰腺癌细胞的定向运动和 ERM 支架

基本信息

批准号：
8779713
负责人：
Kathleen M Mulder
金额：
$ 16.64万
依托单位：
PENNSYLVANIA STATE UNIV HERSHEY MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-01-01 至 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8779713
关键词：
3-Dimensional A kinase anchoring protein Actin-Binding Protein Actins Address Adenocarcinoma Cell Behavior Binding Binding Proteins Biosensor Cause of Death Cell membrane Cell physiology Cells Complex Cyclic AMP-Dependent Protein Kinases Cytoskeleton Development Diagnosis Disease Disease Progression Distant Distant Metastasis Environment Epithelial Cells Event Extracellular Matrix Degradation Extracellular Matrix Proteins Family Fibroblasts Fibronectins Focal Adhesions Growth Human Interphase Cell Intraepithelial Neoplasia Invaded Label Lead Life Link Malignant Epithelial Cell Malignant Neoplasms Malignant neoplasm of pancreas Mediating Membrane Mitogen-Activated Protein Kinases Modeling Molecular Mutation Neoplasm Metastasis Pancreas Pancreatic Ductal Adenocarcinoma Pancreatic carcinoma Patients Play Population Process Production Property Protein Family Proteins Recruitment Activity Regulation Role Scaffolding Protein Signal Pathway Signal Transduction Site Staging Stress Fibers Testing Therapeutic Time Tumor Cell Invasion Tumorigenicity Validation base cancer cell cell motility cellular imaging cohort ezrin genetic regulatory protein in vitro Model in vivo in vivo Model inhibitor/antagonist knock-down member migration moesin molecular dynamics mutant neoplastic cell novel novel therapeutic intervention pancreatic neoplasm protein complex public health relevance radixin protein rapid growth rho scaffold stellate cell targeted treatment tumor tumor growth

项目摘要

DESCRIPTION (provided by applicant): Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies, with median survival times of less than a year. Activating KRas mutations are present at diagnosis in up to 95% of patients, with alterations already existing in 92% of early-stage pancreatic intraepithelial neoplasms. In addition, PDAC displays rapid invasion locally, in addition to distant metastasis, processes that require multiple molecular events that are critical to target therapeutically. While the mode of invasion varies among different PDAC cells (ie, single-cell, collective), cells at the leading edge of invasion zones may develop pathfinder cell functions, which can create the microtracks through which the tumor cells will spread, by degradation of the extracellular matrix (ECM). Actin-rich membrane protrusions (invadopodia) develop in specific cell populations, which lead the migratory cohort of invading cells. Thus, it is critical to understand the mechanisms involved in the formation of these pathfinder cells and the molecular dynamics that occur at the invading edges. Among these events, mutant KRas is required for the formation of invadopodia, and specific cellular scaffolds are required for mediating the effector signaling pathways involved. For example, Ezrin, a scaffold protein that links the plasma membrane to the actin cytoskeleton, as well as other members of the ezrin-radixin-moesin (ERM) family, are present in actin-rich invadopodia and play important roles in PDAC migration. In addition, discrete pools of active RhoA at the leading and lagging edges of motile PDACs are critical for invasive behavior. We have described a novel anti-motility target (km23-1), the depletion of which was shown to diminish Ras and ERK activity, as well as TGFss1 production and cancer cell motility and invasion. It also interacts with numerous actin-regulatory proteins and can modulate RhoA activity and Ezrin expression. We hypothesize that km23-1 plays an important role in the formation and activity of invadopodia by facilitating the assembly of Ezrin- enriched complexes that further regulate Ras activity and protein kinase A (PKA)-modulated RhoA protrusion- retraction events. Therefore, in Aim 1, we will examine the ability of km23-1 depletion to inhibit PDAC invasion in organotypic models representative of the in vivo setting, which will include stromal pancreatic stellate cells. Additional studies will address the functional role of km23-1 in regulating invadopodia RhoA activity and leader cell migration. Further, since ERM and actin are required for Ras activation, we will test the hypothesis that km23-1 regulates Ezrin scaffolded complexes to control Ras activation and downstream events involved in invadopodia protrusion during PDAC invasion. In Aim 2, we will examine the effect of km23-1 depletion on tumorigenicity and metastasis in vivo to determine the potential utility of km23-1 inhibitors as anti-metastatic therapy. The results of these studies will provide significant new information regarding the spatial control of Ras and RhoA activity, as well as Ezrin/actin scaffolding, in PDAC invasion and metastasis. They will also facilitate efforts to develop novel km23-1- and Ezrin-based inhibitors for the treatment of PDAC.

描述（由申请人提供）：胰腺导管腺癌（PDAC）是最致命的恶性肿瘤之一，中位生存时间不到一年。在诊断中有多达95％的患者存在激活KRAS突变，其中92％的早期胰腺上皮内肿瘤已经存在改变。此外，除远处转移外，PDAC还在本地显示快速入侵，这些过程需要多个分子事件，这些事件对于靶向治疗至关重要。虽然侵袭模式在不同的PDAC细胞（即单细胞，集体）之间有所不同，但在入侵区的前沿细胞可能开发探路者细胞功能，可以通过降解细胞外基质（ECM）来创建微裂片，通过该功能通过该功能传播肿瘤细胞。富含肌动蛋白的膜突起（Invadopodia）在特定的细胞群体中发展，这导致了入侵细胞的迁移群体。因此，了解这些探路剂细胞形成的机制以及在入侵边缘处发生的分子动力学。在这些事件中，突变的KRA需要形成Invadopodia，并且需要特定的细胞支架来介导涉及的效应子信号通路。例如，将质膜与肌动蛋白细胞骨架以及Ezrin-radixin-Moesin（ERM）家族的其他成员之间联系起来的Ezrin，一种将质膜连接起来的支架蛋白，在富含肌动蛋白的Invadopodia中存在着重要的作用，并在PDAC迁移中起重要作用。此外，在Motile PDAC的领先和滞后边缘的主动RhoA的离散池对于侵入性行为至关重要。我们已经描述了一种新型的抗动力靶标（KM23-1），其耗竭显示会减少RAS和ERK活性，以及TGFSS1的产生以及癌细胞的运动性和入侵。它还与许多肌动蛋白调节蛋白相互作用，并可以调节RhoA活性和Ezrin表达。我们假设KM23-1通过促进Ezrin-富含Ezrin-富集的复合物的组装，在Invadopodia的形成和活性中起着重要作用，从而进一步调节RAS活性和蛋白激酶A（PKA）调节的RhoA rhoA protrusion-Retraction-Retraction-Retraction-Retraction-Retraction-Retraction-Retraction-Retraction-rase a（PKA）。因此，在AIM 1中，我们将检查KM23-1耗竭抑制代表体内环境的器官型模型中PDAC侵袭的能力，其中包括基质胰腺星状细胞。其他研究将解决KM23-1在调节Invadopodia RhoA活性和领导者细胞迁移中的功能作用。此外，由于RAS激活需要ERM和肌动蛋白，因此我们将测试以下假设：KM23-1调节Ezrin脚手架的复合物以控制PDAC入侵期间与Invadopodia togrusion有关的RAS激活和下游事件。在AIM 2中，我们将研究KM23-1耗竭对体内肿瘤性和转移的影响，以确定KM23-1抑制剂作为抗中转移疗法的潜在效用。这些研究的结果将为PDAC入侵和转移提供有关RAS和RHOA活性的空间控制以及Ezrin/肌动蛋白支架的大量新信息。他们还将促进开发新型KM23-1-和Ezrin基于Ezrin的抑制剂以治疗PDAC。