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

CCR4-NOT复合物在核孔复合物组装调节中的作用

• 批准号: 10665063
• 负责人: Stephen Sakuma
• 金额: $ 3.53万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-03-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10665063
• 关键词: 3-Dimensional; Biological Assay; Cancer Biology; Cause of Death; Cell Death; Cell Nucleus; Cell Proliferation; Cell physiology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Complex; Environment; Eukaryota; Flow Cytometry; Genes; Genetic; Genetic Transcription; Goals; Imaging Techniques; Immune; Immunooncology; Immunosuppression; Individual; Leukocytes; Macrophage; Malignant Neoplasms; Measurement; Measures; Mediating; Messenger RNA; Metabolism; Methods; Molecular; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Pathway Analysis; Pathway interactions; Patients; Phase; Phenotype; Postdoctoral Fellow; Proliferating; Proteins; Regulation; Regulator Genes; Research; Research Project Grants; Role; Severities; Small Interfering RNA; Stains; Structure; Surface; Survival Rate; T-Lymphocyte; Therapeutic; Toxic effect; Translational Repression; Tumor-associated macrophages; Work; addiction; cancer cell; cancer immunotherapy; cancer therapy; cell type; clinical efficacy; effective therapy; experimental study; immune checkpoint blockade; immune checkpoint blockers; knock-down; knockout gene; mRNA Transcript Degradation; member; monocyte; neoplastic cell; novel; nucleocytoplasmic transport; programs; protein structure; response; side effect; single-cell RNA sequencing; stem; success; tumor; tumor growth; tumor microenvironment; tumor xenograft; tumor-immune system interactions; tumorigenic; whole genome

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.

项目概要 癌症仍然是全世界死亡的主要原因。尽管付出了巨大努力 尽管正在开发有效的治疗方法，但许多类型的肿瘤的存活率仍然很低。长期目标是 这项工作的 F99 和 K00 阶段都将揭示导致新型抗癌疗法的新策略。 F99 阶段将重点关注癌症生物学的细胞内在方面，而 K00 阶段将重点关注癌症生物学的细胞内在方面。 免疫细胞具有直接抑制肿瘤生长和调节已批准药物疗效的巨大潜力 癌症免疫疗法。 F99阶段：越来越多的证据表明许多癌细胞会产生成瘾性 核运输机械并增加了核孔复合体（NPC）的数量， 调节分子核细胞质运输的蛋白质结构。最近的工作表明 抑制 NPC 的组装可能是癌症治疗的有效靶标。然而， 调节 NPC 组装的分子因素尚未完全了解，并且尚未找到可靶向的途径 被识别。进行全基因组 siRNA 筛选以寻找调节这些组装的基因 结构。经鉴定，当基因被敲除时，会增加或减少 NPC 的含量。 核表面。研究发现敲低 CCR4-NOT 复合体的 3 个成员会增加 NPC。这 CCR4-NOT 是一种主要的去腺苷酸酶复合物，通过调节 mRNA 代谢来控制蛋白质水平 真核生物。这里提出的工作将描述 CNOT 击倒对 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学。此外，这项工作将阐明 CCR4-NOT 复合物调节 NPC 的分子机制。 K00阶段：近年来，免疫肿瘤疗法，即免疫检查点阻断剂，已 表现出显着的临床疗效，但只有一小部分患者对治疗有反应。中的一个 缺乏疗效的原因源于强免疫抑制肿瘤的存在 微环境（TME）。在 TME 被修改成更具免疫激活的环境之前，免疫 检查点封锁疗法的成功有限。该项目的 K00 阶段将重点关注独特的 动态白细胞对观察到的免疫抑制有很大贡献；肿瘤相关 巨噬细胞（TAM）。 TAM 已被证明占肿瘤质量的 50%，并且具有亲和性和亲和性。 抗肿瘤发生表型。仍然需要开发更有效的策略来针对这种细胞类型和 重新编程。 K00 研究将描述可用于调节的转录网络的特征 TME 中巨噬细胞复极化。 F99阶段的成功完成将揭示癌症的可靶向细胞内在方面 生物学，而 K00 阶段的完成将揭示免疫肿瘤学的靶向途径。