Role of Nuclear Pore-Regulated Mechanisms in Prostate Cancer Aggressiveness

项目总结 晚期癌症是人类最致命的疾病之一，是转移性前列腺癌。 经典的例子。尽管有目前的治疗方法，转移性PC进展到耐药阶段， 先于杀伤力。因此，有必要确定导致癌症的新的靶向机制。 积极进取，继续改善患者的预后。这项研究提案的主要目的是 剖析核孔复合体(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调控的靶向通路 致命的个人电脑。核祖细胞通过其构建的核孔蛋白(NUP)发挥不同的生物学功能， 包括核质运输、染色质组织、基因表达、基因组完整性和有丝分裂 监管。然而，在癌症的背景下，核素与肿瘤的形成和发展有关。 它们在癌症发病机制中的具体作用在很大程度上仍不清楚。最近我们在临床上鉴定出 通过询问公开可获得的患者组织样本在致死性PC中上调相关核蛋白 包含原发和对去势化疗耐药的转移性前列腺癌的转录数据集。 在上调的核蛋白中，POM121被证明在致死性PC的侵袭性(细胞存活， 增殖和肿瘤发生)通过核运输调节。然而，具体的交通依赖和- POM121调节PC的攻击性特征及其与其他设备的串扰的独立机制 核子核仍然是未知的。值得注意的是，功能基因组研究(转录学和计算分析) 结合生化和单细胞高分辨率成像，揭示了两个顶部之间的串扰 在致死性PC、POM121和TPR中上调NUPS，从而影响PC细胞的侵袭性。 具体地说，我们发现PC细胞的存活通过特定的POM121转录调节而增强 基因组稳定性基因和TPR，TPR通过雄激素受体调节有丝分裂检查点活动 (Ar)依赖和独立的机制。至关重要的是，我们的研究还发现了一个潜在的新角色 可溶染色质结合核蛋白对PC细胞侵袭性特征的调节可能通过直接 转录调控。因此，总而言之，这些结果导致了npc调节pc的假设 通过控制基因组功能和稳定性的多方面机制具有侵略性。我们会 解决这一假设，并通过以下方式确定导致PC攻击性的NUP特定机制 三个目标。在目标1中，我们将定义POM121核运输依赖和独立机制 促进个人电脑的进攻性。在目标2中，我们将研究POM121-TPR机制的相互作用增强 致死性前列腺癌的基因组稳定性和细胞存活率。在目标3中，我们将评估其临床意义和疗效。 在患者衍生的临床前模型中，针对特定的NPC调控的控制基因组完整性的途径。

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.