A Novel Anoikis Effector that Drives Ovarian Cancer Metastasis

一种驱动卵巢癌转移的新型失巢效应器

基本信息

批准号：
10117214
负责人：
Romi Gupta
金额：
$ 7.43万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-02 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10117214
关键词：
American Cancer Society Anchorage-Independent Growth Anoikis Apoptosis Apoptotic Biochemical Genetics Cancer Patient Cell Culture Techniques Cell Transplantation Clinical Data Dependence Diagnosis Disease Disseminated Malignant Neoplasm Drug Targeting Extracellular Matrix Genes Genetic Genome Goals Growth Human Immune system Liver Lung Malignant Neoplasms Malignant neoplasm of ovary Mediator of activation protein Metastatic Neoplasm to the Liver Metastatic Neoplasm to the Lung Metastatic to Methods Modeling Molecular Neoplasm Metastasis Normal Cell Organ Organ Model Ovary Pathway interactions Patients Pharmacology Phenotype Prevalence Prognosis Progression-Free Survivals Public Health RNA interference screen Regulator Genes Regulatory Pathway Repression Research Proposals Resistance Role Sampling Serous Testing Therapeutic TimeLine Transplantation United States Woman Xenograft Model Xenograft procedure base cancer cell cancer therapy cell growth effective therapy experimental study genetic approach in vivo in vivo Model knock-down mouse model neoplastic cell new therapeutic target novel novel therapeutics overexpression prevent small hairpin RNA small molecule inhibitor therapeutic target therapeutically effective tumor growth

项目摘要

PROJECT SUMMARY Normal cells are dependent upon the extracellular cell matrix (ECM) for survival, and undergo apoptosis when they lose contact with the ECM – a phenomenon termed anoikis. The acquisition of anoikis resistance is a critical step that contributes prominently to the metastatic progression in ovarian cancer. However, the factors and regulatory pathways that confer anoikis resistance in ovarian cancer remain largely unknown. Thus, to identify factors that confer anoikis resistance, we performed an unbiased druggable genome-based RNAi screen and identified ATAD2 as a novel factor that confers anoikis resistance in high-grade serous ovarian cancer (HGS- OvCa) cells. Additionally, we document that ATAD2 is overexpressed in HGS-OvCa patient samples and its overexpression predicts significantly reduced overall survival (OS) and progression-free survival (PFS). Our preliminary data that makes the basis of this research proposal provides strong evidence supporting the scientific premise that ATAD2 is necessary for anoikis resistance and thus is a potential driver of HGS-OvCa metastasis. The overall objective for this research proposal is to determine the in vivo role of ATAD2 in facilitating HGS- OvCa metastasis, understand its mechanism-of-action and evaluate in vivo pharmacological targeting of ATAD2 for metastatic HGS-OvCa therapy. Specifically, Aim 1 experiments will determine the role of ATAD2 as a driver of HGS-OvCa metastasis and evaluate it as a drug target for metastatic HGS-OvCa cancer therapy. We will use both genetic approach and highly-effective and selective ATAD2 small molecule inhibitor BAY-850-based pharmacological approach for achieving the goals of Aim 1. For genetic approach, we will use complementary organ-specific and spontaneous metastasis models based on orthotopic xenograft of HGS-OvCa cells. For pharmacological approach, in addition to organ-specific and spontaneous mouse models of HGS-OvCa metastasis, we will also use a novel humanized HGS-OvCa xenograft model with intact innate and adoptive human immune system. Aim 2, experiments will determine the mechanism by which ATAD2 confers anoikis resistance and promote HGS-OvCa metastasis. In preliminary studies, we found that ATAD2 represses the expression of pro-apoptotic gene BAD, which is necessary for ATAD2 inhibition-induced anoikis resistance. Based on these results, we will ascertain the role of BAD as a downstream mediator of ATAD2-indued anoikis resistance and HGS-OvCa metastasis. The experimental approaches will utilize biochemical, genetic, cell culture-based methods and in vivo mouse model of anoikis resistance and HGS-OvCa metastasis. In particular, we will use organ-specific and spontaneous metastasis model based on orthotopic HGS-OvCa cells transplant. Collectively, these results will identify a novel druggable dependency pathway that via promoting anoikis resistance facilitates HGS-OvCa metastasis, and thus can be targeted for effectively treating highly aggressive metastatic HGS-OvCa.

项目摘要正常细胞取决于细胞外细胞基质（ECM）的生存，并在他们失去了与ECM的接触 - 这种现象称为Anoikis。抗药抗性的获取是关键促进卵巢癌转移性进展的步骤。但是，因素和卵巢癌中的抗厌食症会议的监管途径在很大程度上尚不清楚。那是为了识别遇到Anoikis耐药性的因素，我们进行了公正的可吸毒基因组RNAI筛查和鉴定为ATAD2是一个新的因素，它承认高级浆液卵巢癌（HGS- OVCA）细胞。此外，我们记录了ATAD2在HGS-OVCA患者样本中过表达过表达预测显着降低了总生存率（OS）和无进展生存率（PFS）。我们的这项研究基础的初步数据提供了支持科学的有力证据前提是ATAD2对于Anoikis耐药性是必要的，因此是HGS-OVCA转移的潜在驱动力。该研究建议的总体目标是确定ATAD2在促进HGS-的体内作用 OVCA转移，了解其行动机理并评估ATAD2的体内药物靶向用于转移性HGS-ovca疗法。具体而言，AIM 1实验将确定ATAD2作为驱动程序的作用 HGS-ovca转移，并将其评估为转移性HGS-ovca癌症治疗的药物靶标。我们将使用遗传方法以及高效和选择性的ATAD2小分子抑制剂Bay-850 实现目标1的药理方法。对于遗传方法，我们将使用完善基于HGS-OVCA细胞的原位异种移植的器官特异性和赞助转移模型。为了除HGS-OVCA的器官特异性和赞助小鼠模型外，药理学方法转移，我们还将使用具有完整的先天和自适应的新型人性化的HGS-ovca Xenographic模型人类免疫系统。 AIM 2，实验将确定ATAD2承认Anoikis的机制抗性并促进HGS-ovca转移。在初步研究中，我们发现ATAD2繁殖促凋亡基因不良的表达，这对于ATAD2抑制诱导的厌氧性是必不可少的。基于这些结果，我们将确定Bad作为ATAD2指定Anoikis的下游介体的作用抗性和HGS-ovca转移。实验方法将利用生化，遗传，细胞基于培养的方法和Anoikis抗性和HGS-ovca转移的体内小鼠模型。尤其，我们将使用基于原位HGS-OVCA细胞移植的器官特异性和赞助转移模型。总的来说，这些结果将确定一种通过促进Anoikis的新型可吸毒依赖途径抗性促进HGS-OVCA转移，因此可以针对有效治疗高度侵略性转移性HGS-OVCA。