Role of nuclear pore-regulated mechanisms in prostate cancer aggressiveness

项目摘要 晚期癌症是最致命的人类疾病之一，是转移性前列腺癌（PC）， 典型的例子尽管目前的治疗，转移性PC进展到治疗抗性阶段， 会导致死亡因此，有必要确定新的可靶向机制， 积极进取，继续改善患者的预后。本研究计划的主要目的是 剖析核孔复合物（NPC）调节基因组功能（基因表达）的机制 和完整性），并使用创新的临床前模型发现新的NPC调节的靶向途径 致命PC NPC通过它们的结构单元核孔蛋白（NUPS）贡献于不同的生物学功能， 包括核质转运、染色质组织、基因表达、基因组完整性和有丝分裂 调控然而，在癌症的背景下，Nups与肿瘤的形成和发展有关， 它们在癌症发病机理中的具体机制作用仍然很大程度上未知。最近我们在临床上发现 通过询问公开可用的患者组织样品，在致死PC中上调的相关Nups 包含原发性和去势化疗抗性转移性前列腺肿瘤的转录组学数据集。 在上调的Nups中，POM 121显示在致死PC的侵袭性（细胞存活， 增殖和肿瘤发生）。然而，具体的运输依赖和- POM 121调节PC的攻击性特征及其与其他设备的串扰的独立机制 Nups仍然未知。值得注意的是，功能基因组研究（转录组学和计算分析） 结合生物化学和单细胞高分辨率成像，揭示了两个顶级细胞之间的串扰。 在致死PC、POM 121和TPR中上调Nups，这影响PC细胞的攻击性特性。 具体地说，我们发现PC细胞的存活率是通过POM 121特异性转录调控来增强的。 基因组稳定性基因和TPR，TPR又通过雄激素受体调节有丝分裂检查点活性， （AR）依赖性和独立性机制。至关重要的是，我们的研究还发现了一种潜在的新作用， 可溶性染色质结合的Nups在PC细胞的侵袭性特征的调节中可能通过直接的 转录调控因此，总的来说，这些结果导致了这样的假设，即NPC调节PC 通过控制基因组功能和稳定性的多方面机制来控制侵略性。我们将 解决这一假设，并通过以下方式确定促进PC攻击性的NUP特异性机制： 三个目标。在目标1中，我们将定义POM 121核转运依赖性和非依赖性机制 提升PC的攻击性。在目标2中，我们将研究POM 121-TPR机制相互作用增强 基因组稳定性和细胞存活。在目标3中，我们将评估 靶向患者来源的临床前模型中控制基因组完整性的特定NPC调节途径。

PROJECT SUMMARY Advanced carcinomas are among the most lethal human diseases, being metastatic prostate cancer (PC) a classic example. Despite current treatments, metastatic PC progresses to a therapy resistant stage that precedes lethality. Therefore, there is a need to identify new targetable mechanisms driving cancer aggressiveness to continue improving patients’ outcome. The main objective of this research proposal is to dissect the mechanisms by which Nuclear Pore Complexes (NPCs) regulate genome functions (gene expression and integrity) in PC and uncover novel NPC-regulated targetable pathways using innovative pre-clinical models of lethal PC. NPCs contribute to different biological functions through their building blocks nucleoporins (Nups), including nucleo-cytoplasmic transport, chromatin organization, gene expression, genome integrity and mitotic regulation. In the context of cancer, Nups have been associated to tumor formation and development, however their specific mechanistic role in cancer pathogenesis remains largely unknown. Recently we identified clinically relevant Nups upregulated in lethal PC through interrogation of publicly available patient tissue sample transcriptomic datasets containing primary and castration-chemotherapy-resistant metastatic prostate tumors. Within the upregulated Nups, POM121 was shown to play a role in the aggressiveness of lethal PC (cell survival, proliferation and tumorigenesis) via nuclear transport regulation. Yet, the specific transport-dependent and - independent mechanisms by which POM121 regulates the aggressive features of PC and its crosstalk with other Nups remain unknown. Notably, functional genomic studies (transcriptomic and computational analysis) combined with biochemical and single-cell high-resolution imaging have revealed a crosstalk between two top upregulated Nups in lethal PC, POM121 and TPR, which impacts on the aggressiveness properties of PC cells. Specifically, we found that PC cell survival is enhanced by the POM121 transcriptional regulation of specific genome stability genes and TPR, which in turn regulates mitotic checkpoint activity, through Androgen Receptor (AR)-dependent and independent mechanisms. Crucially, our studies have also uncovered a potential novel role of soluble chromatin-bound Nups in the regulation of aggressive features of PC cells potentially through direct transcriptional regulation. Thus, collectively these results led to the hypothesis that NPCs regulate PC aggressiveness through multifaceted mechanisms controlling both genome functions and stability. We will address this hypothesis and determine the Nup-specific mechanisms contributing to PC aggressiveness through three aims. In Aim 1, we will define POM121 nuclear transport-dependent and -independent mechanisms promoting PC aggressiveness. In Aim 2, we will examine the POM121-TPR mechanistic interplay enhancing genome stability and cell survival in lethal PC. In Aim 3, we will evaluate the clinical significance and efficacy of targeting specific NPC-regulated pathways controlling genome integrity in patient derived pre-clinical models.