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: 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
• 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

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.

项目摘要 癌症仍然是全球死亡的主要原因。尽管致力于 在许多类型的肿瘤中开发有效的治疗方法，生存率仍然很低。长期目标 这项工作的F99和K00阶段都将是揭示导致新型抗癌疗法的新策略。 F99阶段将集中在癌症生物学的细胞内部方面，而K00阶段将集中于 免疫细胞具有很大的潜力，可以直接抑制肿瘤生长并调节已经批准的效率 癌症免疫疗法。 F99阶段：越来越多的证据表明，许多癌细胞都会成瘾 到核运输机械，并增加了核孔复合物数量（NPC），多数 调节分子核质转运的蛋白质结构。最近的工作表明 抑制NPC的组装可以代表癌症治疗的有效靶标。但是， 调节NPC组装的分子因子尚不完全了解，并且有目标的途径尚未 被识别。进行了整个基因组siRNA筛选以找到调节这些组装的基因 结构。鉴定出基因，当被击倒时，会增加或减少NPC含量 核表面。发现CCR4-NOT复合物的3个成员的敲低可增加NPC。这 CCR4-not是一种主要的去甲基酶配合物，可以通过调节mRNA代谢来控制蛋白水平 真核生物。这里提出的工作将表征CNOT敲低对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不调节NPC的分子机制。 K00阶段：近年来，免疫肿瘤疗法，即免疫障碍剂，具有 显示出显着的临床效率，但是只有一小部分患者对治疗做出反应。中的一个 缺乏效率的原因是由于存在强烈的免疫抑制肿瘤的存在 微环境（TME）。直到可以将TME修改为更具免疫激活的环境为止 检查点封锁疗法的成功将有限。该项目的K00阶段将集中于独特的 和动态白细胞强烈有助于观察到的免疫抑制；肿瘤相关 巨噬细胞（TAM）。 TAM已显示出多达50％的肿瘤质量，并且具有亲和 抗肿瘤的表型。需要制定更有效的策略来针对这种细胞类型的需求和 重新编程。 K00研究将表征可利用的转录网络 TME中的巨噬细胞复置。 F99阶段的成功完成将揭示癌症的细胞内在方面 生物学，而k00阶段的完成将揭示免疫肿瘤的目标途径。