Spatial regulation of mechanistic target of rapamycin complex 1 (mTORC1) and its role in oral squamous cell carcinoma

雷帕霉素复合物1（mTORC1）机械靶点的空间调控及其在口腔鳞状细胞癌中的作用

• 批准号: 10750317
• 负责人: Ayse Zisan Sahan
• 金额: $ 4.15万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750317
• 关键词: 3-Phosphoinositide Dependent Protein Kinase-1; Affect; Amino Acids; Apoptosis; Autophagocytosis; Biochemical; Biological Assay; Biosensor; Cancer Model; Cell Line; Cell Nucleus; Cell Proliferation; Cell Survival; Cells; Cessation of life; Complex; Cytosol; DNA Sequence Alteration; Data; Disease; Drug resistance; Enzyme-Linked Immunosorbent Assay; Enzymes; Epidermal Growth Factor; FRAP1 gene; Fluorescence Resonance Energy Transfer; Genes; Genetic Transcription; Growth; Growth Factor; In Vitro; Inflammatory; Location; Lysosomes; Malignant Epithelial Cell; Malignant Neoplasms; Metabolism; Monitor; Mus; Nuclear; Nutrient; Pathway interactions; Phosphatidylinositols; Phosphotransferases; Play; Proliferating; Proteins; Reactive Oxygen Species; Regulation; Reporter; Reporting; Research; Resistance; Role; Signal Pathway; Signal Transduction; Specificity; Survival Rate; Testing; Transcriptional Regulation; Translations; Work; Xenograft Model; cell growth; cytokine; differential expression; drug sensitivity; experimental study; in vivo; inhibitor; mouth squamous cell carcinoma; peroxisome; phosphoproteomics; tool; transcription factor; transcriptome sequencing; tumor; tumor growth; tumor xenograft

Project Summary Mechanistic target of Rapamycin (mTOR) complex 1 (mTORC1) integrates inputs from multiple pathways and senses diverse signals to regulate cell growth, protein translation, and proliferation1,2. Given that spatial compartmentalization can enhance signaling specificity and efficiency3,4, spatial regulation of mTORC1 appears to be critical for this multifaceted signaling complex5 as it has been reported at many subcellular locations6–9. For example, mTORC1 at the lysosome is regulated by both amino acids and growth factors and functions to promote translation and suppress autophagy10. The complex has also been identified at peroxisomes, where mTORC1 responds to reactive oxygen species7. However, the roles of mTORC1 at other subcellular locations where its presence has been reported, in particular in the nucleus, are not well understood. A major limitation in the field is the availability of tools to assess the activity and function of spatially compartmentalized signaling enzymes in living cells. Using a genetically encoded fluorescent biosensor to monitor mTORC1 activity, the Zhang lab has previously discovered a pool of nuclear mTORC1, which has yet to be defined in function. Based on our preliminary data, we hypothesize that nuclear mTORC1 regulates pro-inflammatory transcription. We will test this hypothesis by combining targeted inhibition of mTORC1 with a phosphoproteomics experiment and biochemical assays. Lastly, since genetic mutations activating phosphoinositide 3-kinase(PI3K)/protein kinase B (Akt)/mTOR signaling are prominent alterations in oral squamous cell carcinoma (OSCC)11,12 and mTORC1 inhibition can induce tumor regression13,14, we will study the role of subcellular mTORC1 signaling in the growth and drug resistance of OSCC cells in vitro and in vivo in mice. We hypothesize that inhibiting mTORC1 in the nucleus or the cytosol will differentially effect cell growth and EGFR inhibitor resistance in OSCC. The proposed project will elucidate the function of the previously undefined pool of nuclear mTORC1 and clarify the roles of subcellular mTORC1 in the growth and drug resistance of OSCC.

项目摘要 雷帕霉素（MTOR）复合物1（MTORC1）的机械靶标会整合来自多个途径的输入 感觉潜水员信号以调节细胞生长，蛋白质翻译和增殖1,2。鉴于该空间 隔室化可以提高信号传导特异性和效率3,4，MTORC1的空间调节出现 对于这种多面信号传导复合物至关重要，因为在许多亚细胞位置已有报道6-9。 例如，溶酶体处的mTORC1受氨基酸和生长因子和功能的调节 促进翻译并抑制自噬10。该络合物也已在过氧化物体中鉴定 MTORC1对活性氧响应7。但是，MTORC1在其他亚细胞位置的作用 据报道，尤其是在细胞核中的存在，尚不清楚。一个主要限制 该领域是评估空间分隔信号的活动和功能的工具的可用性 活细胞中的酶。使用一般编码的荧光生物传感器监测MTORC1活性， 张实验室以前已经发现了一个核MTORC1库，该核MTORC1尚未在功能中定义。基于 在我们的初步数据上，我们假设核MTORC1调节促炎性转录。我们将 通过将靶向抑制MTORC1与磷蛋白质组学实验相结合，检验该假设 生化测定。最后，由于遗传突变激活磷酸肌醇3-激酶（PI3K）/蛋白激酶 B（AKT）/MTOR信号传导是口服鳞状细胞癌（OSCC）11,12和MTORC1的显着改变 抑制作用可以诱导肿瘤回归13,14，我们将研究亚细胞MTORC1信号在生长中的作用 OSCC细胞在体外和小鼠体内的耐药性。我们假设抑制MTORC1 OSCC中的细胞核或细胞质会对细胞生长和EGFR抑制剂的耐药性不同。提议 项目将阐明先前未定义的核MTORC1库的功能，并阐明 OSCC生长和耐药性的亚细胞MTORC1。