The role of the CCR4-NOT complex in the regulation of Nuclear Pore Complex assembly

项目总结 癌症仍然是世界范围内主要的死亡原因。尽管付出了巨大的努力来 在开发有效的治疗方法后，许多类型的肿瘤的存活率仍然很低。的长期目标是 这项工作的F99和K00阶段都将揭示导致新的抗癌疗法的新策略。 F99阶段将专注于癌症生物学的细胞固有方面，而K00阶段将专注于 具有巨大潜力的免疫细胞直接抑制肿瘤生长并调节已获批准的疗效 癌症免疫疗法。 F99期：越来越多的证据表明，许多癌细胞会上瘾 对于核运输机械，并增加了核孔复合体(NPC)的数量，多- 调节核质分子运输的蛋白质结构。最近的研究表明， 抑制神经前体细胞的组装可能是癌症治疗的有效靶点。然而， 调控鼻咽癌组装的分子因素还不完全清楚，还没有一个有针对性的途径 被指认出来。进行了全基因组siRNA筛选以寻找调节这些基因组装的基因 结构。已确定的基因，当被击倒时，会增加或减少鼻咽癌细胞上的NPC含量 原子核表面。敲除CCR4-NOT复合体的3个成员可增加NPC。这个 CCR4-NOT是一种主要的死烯基酶复合体，通过调节mRNA的代谢来控制蛋白质水平。 真核生物。这里提出的工作将表征CNOT基因敲除对NPC的影响 数量/功能以及这些变化如何改变基本的细胞生理。此外，这项工作将阐明 CCR4-NOT复合体调节鼻咽癌的分子机制。 K00阶段：近年来，免疫肿瘤学治疗，即免疫检查点阻滞剂， 显示出显着的临床疗效，但只有一小部分患者对治疗有反应。其中一个 缺乏疗效的原因源于一种强烈的免疫抑制肿瘤的存在 微环境(TME)。直到TME可以被改造成更具免疫活性的环境，免疫 检查站封锁疗法的成功将是有限的。该项目的K00阶段将专注于一个独特的 以及对观察到的免疫抑制有强烈贡献的动态白细胞；与肿瘤相关的 巨噬细胞()。TAMs已被证明构成高达50%的肿瘤质量，并且既有促癌作用，也有促癌作用 抗肿瘤表型。仍然需要开发更有效的策略来针对这种类型的细胞和 重新编程。K00的研究将描述可以被利用来调节的转录网络的特征 TME内巨噬细胞复极。 F99期的成功完成将揭示癌症的靶向细胞--内在方面 生物学，而K00阶段的完成将揭示一条免疫肿瘤学靶向途径。

PROJECT SUMMARY Cancer continues to be a leading cause of death worldwide. Despite significant efforts devoted towards developing effective treatments, the survival rates remain low in many types of tumors. The long-term goal of both the F99 and K00 phases of this work will be to reveal new strategies that lead to novel anti-cancer therapies. The F99 phase will be focused on a cell-intrinsic aspect of cancer biology, while the K00 phase will focus on an immune cell with great potential to directly inhibit tumor growth and to modulate the efficacy of already approved cancer immunotherapies. The F99 phase: An increasing body of evidence has shown that many cancer cells develop an addiction to the nuclear transport machinery and have increased numbers of Nuclear Pore Complexes (NPCs), the multi- protein structures that regulate the nucleocytoplasmic transport of molecules. Recent work has revealed that inhibition of the assembly of NPCs could represent an effective target for cancer therapeutics. However, the molecular factors that regulate NPC assembly are incompletely understood and a targetable pathway has yet to be identified. A whole genome siRNA screen was performed to find genes that regulate assembly of these structures. Genes were identified that, when knocked down, either increase or decrease the NPC content on the surface of the nucleus. Knockdown of 3 members of the CCR4-NOT complex were found to increase NPCs. The CCR4-NOT is a major deadenylase complex that controls protein levels via regulation of mRNA metabolism in eukaryotes. The work proposed here will characterize the effect of CNOT knockdown on the NPC number/functionality and how these changes alter basic cellular physiology. Additionally, this work will elucidate the molecular mechanism by which the CCR4-NOT complex regulates NPCs. The K00 phase: In recent years immuno-oncology therapies, namely immune checkpoint blockers, have shown remarkable clinical efficacy however only a small proportion of patients respond to treatments. One of the reasons underlying the lack of efficacy stems from the presence of a strong immunosuppressive tumor microenvironment (TME). Until the TME can be modified into a more immuno-activating environment, immune checkpoint blockade therapies will have limited success. The K00 phase of this project will focus on a unique and dynamic leukocyte that strongly contributes to the observed immunosuppression; the Tumor-Associated Macrophage (TAM). TAMs have been shown to compose up to 50% of a tumor’s mass and have both pro- and anti-tumorigenic phenotypes. The need remains to develop more effective strategies to target this cell type and reprogram it. The K00 research will characterize the transcriptional networks that can be harnessed to modulate macrophage repolarization in the TME. Successful completion of the F99 phase will reveal a targetable cell-intrinsic aspect of cancer biology, while completion of the K00 phase will reveal an immuno-oncology targetable pathway.