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.

项目概要正常细胞的生存依赖于细胞外细胞基质（ECM），当细胞外基质（ECM）存在时，就会发生细胞凋亡。他们与 ECM 失去联系——这种现象被称为失巢凋亡。获得失巢凋亡抵抗力是一个关键。然而，影响卵巢癌转移进展的因素和因素。因此，在卵巢癌中赋予失巢凋亡抗性的调控途径仍然很大程度上未知。为了确定赋予失巢凋亡抗性的因素，我们进行了无偏倚的基于可药物基因组的 RNAi 筛选，确定 ATAD2 是一种赋予高级别浆液性卵巢癌失巢凋亡抗性的新因子（HGS- 此外，我们记录了 ATAD2 在 HGS-OvCa 患者样本及其中过度表达。过度表达预示着总生存期 (OS) 和无进展生存期 (PFS) 显着降低。构成本研究提案基础的初步数据提供了支持科学的有力证据前提是 ATAD2 对于失巢凋亡抵抗是必需的，因此是 HGS-OvCa 转移的潜在驱动因素。本研究提案的总体目标是确定 ATAD2 在促进 HGS- OvCa 转移，了解其作用机制并评估 ATAD2 的体内药理学靶向具体来说，Aim 1 实验将确定 ATAD2 作为驱动因素的作用。 HGS-OvCa 转移的研究，并评估其作为转移性 HGS-OvCa 癌症治疗的药物靶点。兼具基因方法和基于 BAY-850 的高效选择性 ATAD2 小分子抑制剂实现目标 1 目标的药理学方法。对于遗传方法，我们将使用补充基于 HGS-OvCa 细胞原位异种移植的器官特异性和自发转移模型。除了 HGS-OvCa 的器官特异性和自发小鼠模型之外，药理学方法转移，我们还将使用具有完整先天性和过继性的新型人源化 HGS-OvCa 异种移植模型目标 2，实验将确定 ATAD2 赋予失巢凋亡的机制。在初步研究中，我们发现 ATAD2 会抑制 HGS-OvCa 的耐药性并促进 HGS-OvCa 转移。促凋亡基因 BAD 的表达，这是 ATAD2 抑制诱导的失巢凋亡抵抗所必需的。基于这些结果，我们将确定 BAD 作为 ATAD2 引起的失巢凋亡下游介质的作用耐药性和 HGS-OvCa 转移实验方法将利用生化、遗传、细胞。基于培养的方法和失巢凋亡抵抗和 HGS-OvCa 转移的体内小鼠模型。我们将使用基于原位HGS-OvCa细胞移植的器官特异性和自发转移模型。总的来说，这些结果将确定一种新的药物依赖性途径，通过促进失巢凋亡耐药性促进 HGS-OvCa 转移，因此可以有针对性地有效治疗高度侵袭性转移性 HGS-OvCa。