A Novel Anoikis Effector that Drives Ovarian Cancer Metastasis

项目总结 正常细胞依赖细胞外基质(ECM)生存，并在以下情况下发生凋亡 他们与ECM失去联系--一种被称为失巢的现象。获得失巢抵抗是一个关键 在卵巢癌的转移进展中起重要作用的步骤。然而，这些因素和 卵巢癌中诱导失巢细胞耐药的调控途径在很大程度上仍不清楚。因此，要确定 ，我们进行了基于基因组的无偏向药物RNAi筛查，并 发现ATAD2是高级别浆液性卵巢癌(HGS-1)中的一种新的失巢抵抗因子 OvCa)细胞。此外，我们还发现ATAD2在HGS-OvCa患者样本中过表达，其 过度表达可显著降低总生存期(OS)和无进展生存期(PFS)。我们的 构成这项研究建议基础的初步数据提供了强有力的证据，支持科学的 假设ATAD2是失巢耐药所必需的，因此是HGS-OvCa转移的潜在驱动因素。 这项研究的总体目标是确定ATAD2在体内促进HGS- 卵巢肿瘤转移机制的研究及ATAD2体内药理靶向的评价 用于转移性HGS-OvCa治疗。具体地说，Aim 1的实验将确定ATAD2作为驾驶员的角色 并评价其作为HGS-OvCa转移性肿瘤治疗的药物靶点。我们将使用 遗传方法与高效选择性ATAD2小分子抑制剂BAY-850 实现目标的药理学方法1.对于基因方法，我们将使用互补 基于HGS-OvCa细胞异种原位移植的器官特异性和自发性转移模型。为 除器官特异性和自发的HGS-Ovca小鼠模型外的药理学方法 转移，我们还将使用一种新的人源化HGS-OvCa异种移植模型，该模型具有完整的先天和通过 人类的免疫系统。目的2，实验将确定ATAD2赋予失巢细胞的机制 抵抗和促进HGS-OvCa转移。在初步研究中，我们发现ATAD2抑制 促凋亡基因Bad的表达是ATAD2抑制诱导失巢抵抗所必需的。 基于这些结果，我们将确定BAD作为ATAD2诱导的失巢细胞的下游调节因子的作用 耐药与HGS-OvCa转移的关系实验方法将利用生化、遗传、细胞 基于培养的方法和体内小鼠模型的失巢抵抗和HGS-OvCa转移。特别是， 我们将在HGS-OvCa细胞原位移植的基础上，采用器官特异性和自发性转移模型。 总而言之，这些结果将确定一种新的可药物依赖途径，通过促进失巢凋亡 耐药促进HGS-OvCa转移，因此可作为有效治疗高度侵袭性肿瘤的靶向。 转移性HGS-OvCa。

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.