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

基本信息

批准号：
10530225
负责人：
Stephen Sakuma
金额：
$ 3.59万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10530225
关键词：
3-Dimensional Biological Assay Cancer Biology Cause of Death Cell Death Cell Nucleus 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 Lead Leukocytes Malignant Neoplasms Measurement Measures Mediating Messenger RNA Metabolism Methods Molecular Nuclear Pore Complex Nuclear Pore Complex Proteins Pathway Analysis Pathway interactions Patients Phase Phenotype Proliferating Proteins Regulation Regulator Genes Research Research Project Grants Role Severities Small Interfering RNA Stains Structure Surface Survival Rate T-Lymphocyte Therapeutic Toxic effect Translations 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 macrophage member monocyte neoplastic cell novel nucleocytoplasmic transport response side effect single-cell RNA sequencing stem success tumor tumor growth tumor microenvironment tumor xenograft tumor-immune system interactions tumorigenic whole genome

项目摘要

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.

项目概要尽管付出了巨大的努力，癌症仍然是全世界死亡的主要原因。开发有效的治疗方法，许多类型肿瘤的存活率仍然很低。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，最近的工作表明，调节分子核细胞质运输的蛋白质结构。抑制 NPC 的组装可能是癌症治疗的有效靶标。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进行全基因组 siRNA 筛选以寻找调节这些组装的基因。经鉴定，当基因被敲除时，会增加或减少 NPC 的含量。发现 CCR4-NOT 复合体的 3 个成员的敲低会增加 NPC。 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这里提出的工作将描述 CNOT 敲低对 NPC 的影响。此外，这项工作将阐明数量/功能以及这些变化如何改变基本的细胞生理学。 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。缺乏疗效的原因源于强免疫抑制性肿瘤的存在直到TME能够被修改成更具免疫激活的环境、免疫。检查点封锁疗法的成功有限，该项目的 K00 阶段将专注于一种独特的疗法。动态白细胞对观察到的肿瘤相关免疫抑制有很大贡献；巨噬细胞 (TAM) 已被证明占肿瘤质量的 50%，并且具有亲和性。仍然需要开发更有效的策略来针对这种细胞类型和 K00 研究将描述可用于调节的转录网络。 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。