Role of Insulin Receptor Substrates in Kras-driven lung cancer

胰岛素受体底物在 Kras 驱动的肺癌中的作用

• 批准号: 10163809
• 负责人: Nada Y. Kalaany
• 金额: $ 40.49万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10163809
• 关键词: Ablation; Adaptor Signaling Protein; Address; Affect; Apoptosis; Area; Biological Assay; Breeding; Cancer Cell Growth; Cancer Etiology; Cancer Patient; Cancer cell line; Cell Culture Techniques; Cell Cycle Arrest; Cells; Cessation of life; Complex; Cre-LoxP; Critical Pathways; Cultured Cells; Exhibits; FOXO1A gene; FOXO3A gene; FRAP1 gene; Feedback; Gene Activation; Gene Expression; Genes; Genetic; Genetically Engineered Mouse; Glucose; Glycolysis; Goals; Hepatic; Human; IGF1 gene; IRS1 gene; IRS2 gene; Image; Imaging Techniques; Immune; In Vitro; Incidence; Insulin; Insulin Receptor; Insulin-Like Growth Factor I; Investigation; K-ras mouse model; KRAS2 gene; Lung; Lung Neoplasms; Maintenance; Malignant Neoplasms; Malignant neoplasm of lung; Mass Spectrum Analysis; Mediating; Messenger RNA; Metabolic; Metabolic Pathway; Metabolism; Mouse Strains; Mus; Mutation; Non-Small-Cell Lung Carcinoma; Oncogenes; Pathway interactions; Pentosephosphate Pathway; Pharmacology; Protein Analysis; Role; Signal Pathway; Signal Transduction; TP53 gene; Tamoxifen; Therapeutic; Therapeutic Intervention; Toxic effect; Tumor Burden; Ursidae Family; Xenograft procedure; base; cell transformation; combat; experimental study; forkhead protein; glucose metabolism; glucose production; glucose uptake; human disease; in vivo; in vivo imaging; innovation; knock-down; lung tumorigenesis; neoplastic cell; novel; overexpression; protein metabolite; response; restoration; subcutaneous; therapeutic target; therapeutically effective; tumor; tumor growth; tumor initiation; tumor metabolism; tumor progression; tumor xenograft; tumorigenic

PROJECT SUMMARY/ABSTRACT Non-small cell lung cancer (NSCLC) accounts for the majority of lung cancer, which to-date remains the leading cause of cancer death in the U.S. and worldwide. About 25% of NSCLC harbors Kras oncogene activating mutations. Because direct therapeutic targeting of Kras proved challenging, strategies shifted to targeting downstream effector signaling pathways. However, their efficacy and toxicity remain under investigation, and alternative therapeutic approaches are urgently needed. Evidence from pharmacological and cell culture studies point to a role for insulin and insulin-like growth factor-1 (IGF-1) signaling in Kras-driven lung cancer. However, the specific contribution of this pathway to Kras-driven tumor initiation and progression is unclear, and its role in altering lung tumor metabolism is unknown. Most, if not all insulin/IGF-1 signaling in the lungs converges intracellularly onto the adaptor proteins insulin receptor substrates IRS1 and IRS2 prior to diverging to a complex network of downstream signaling effectors, including PI3K/Akt. The forkhead transcription factors Foxo1 and Foxo3 are insulin-regulated targets that are inactivated by Akt, and affect cellular metabolism, survival and proliferation. Foxos are well known for regulating hepatic glucose metabolism by promoting glucose production and suppressing its utilization. However, the potential tumor-suppressing roles of Foxos in Kras-driven cancers have not been investigated. Here, using distinct conditional genetically engineered mouse (GEM) models of Kras-driven lung cancer, the effects of genetic ablation of IRS1 and IRS2 on the initiation, maintenance, and metabolism of lung tumors, as well as the roles of Foxo1 and Foxo3 in mediating these effects, will be investigated. Histopathological and in vivo imaging techniques will be used to assess at timepoints concomitant with, or subsequent to Kras activation, the effect of IRS gene loss on lung tumor latency, tumor burden and survival of these mice and whether additional loss of Foxo1 and Foxo3 genes would reverse such effects. Moreover, the differential activation of signaling pathways and expression of genes that regulate glucose utilization will be assayed for, and mass spectrometry analyses of glucose-derived metabolites will be performed on the tumors upon loss of IRS genes in the presence or absence of the Foxo genes. Cells will also be isolated from the Kras-driven tumors and grown in culture. Approaches similar to the ones described for in vivo tumors will then be performed on the cells in vitro, to identify targets downstream of IRSs and Foxos, that can alter glucose utilization and hence affect lung tumor maintenance. The role of these targets can then be confirmed via knockdown/overexpression studies. In addition, similar studies will be performed in established, Kras-driven, human NSCLC cell lines with stable inducible knockdown of IRS1 and/or IRS2 that can also be grown as subcutaneous xenografts in immune-deficient mice. Results from these studies will reveal novel metabolic Kras-driven lung cancer vulnerabilities that could be exploited therapeutically in NSCLC patients. !

项目摘要/摘要 非小细胞肺癌（NSCLC）是大多数肺癌，迄今为止，这是 美国和全球癌症死亡的主要原因。 NSCLC大约25％的NSCLC KRAS ONCONENE 激活突变。因为将克拉斯的直接治疗靶向被证明是具有挑战性的，所以策略转移到 靶向下游效应器信号通路。但是，它们的功效和毒性仍在 迫切需要研究和替代治疗方法。药理学和 细胞培养研究表明，胰岛素和胰岛素样生长因子-1（IGF-1）信号在KRAS驱动中的作用 肺癌。但是，该途径对KRAS驱动的肿瘤起始和进展的具体贡献 尚不清楚，其在改变肺肿瘤代谢中的作用尚不清楚。大多数（如果不是全部）胰岛素/IGF-1信号传导 肺细胞内收敛到衔接蛋白胰岛素受体底物IRS1和IRS2之前 与包括PI3K/AKT在内的下游信号效应子的复杂网络分歧。叉子 转录因子FOXO1和FOXO3是胰岛素调节的靶标，被AKT灭活，并影响 细胞代谢，生存和增殖。 Foxos以调节肝葡萄糖代谢而闻名 通过促进葡萄糖生产并抑制其利用。但是，潜在的肿瘤抑制 尚未研究Foxos在KRAS驱动的癌症中的作用。在这里，使用不同的条件遗传 KRAS驱动的肺癌的工程小鼠（GEM）模型，IRS1和IRS2的遗传消融的影响 关于肺肿瘤的启动，维护和代谢，以及FOXO1和FOXO3在 将研究这些效果。组织病理学和体内成像技术将用于 与KRAS激活相关的时间点评估IRS基因丧失对肺的影响 这些小鼠的肿瘤潜伏期，肿瘤负担和存活以及FOXO1和FOXO3基因的额外损失是否 会扭转这种影响。此外，信号通路的差异激活和基因的表达 将分析调节葡萄糖利用的调节，并对葡萄糖衍生的质谱分析进行分析 在存在或不存在FOXO的情况下，将在IRS基因丧失后对肿瘤进行代谢产物 基因。细胞也将从KRAS驱动的肿瘤中分离出来，并在培养中生长。与 然后，将在体外对体内肿瘤进行描述的人，以确定下游的目标 IRS和FOXOS，可以改变葡萄糖利用，因此会影响肺部肿瘤的维持。这些的作用 然后可以通过敲低/过表达研究来确认目标。另外，类似的研究将是 以稳定的IRS1的稳定诱导型敲低稳定的KRAS驱动的人NSCLC细胞线进行 和/或IRS2，也可以作为免疫缺陷小鼠中皮下异种移植物生长。这些结果 研究将揭示可以利用的新型代谢KRAS驱动的肺癌脆弱性 在NSCLC患者中进行治疗。 呢