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: 9383140
• 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/9383140
• 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; 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; Treatment Efficacy; Tumor Burden; Tumorigenicity; 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; mouse model; neoplastic cell; novel; overexpression; protein metabolite; response; restoration; subcutaneous; therapeutic target; 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. !

项目概要/摘要 非小细胞肺癌（NSCLC）占肺癌的大部分，迄今为止仍然是肺癌的主要类型。 美国和全世界癌症死亡的主要原因。约 25% 的 NSCLC 含有 Kras 癌基因 激活突变。由于 Kras 的直接治疗靶向被证明具有挑战性，因此策略转向 靶向下游效应信号通路。然而，它们的功效和毒性仍然存在 迫切需要研究和替代治疗方法。来自药理学和 细胞培养研究指出胰岛素和胰岛素样生长因子 1 (IGF-1) 信号在 Kras 驱动的过程中发挥作用 肺癌。然而，该途径对 Kras 驱动的肿瘤发生和进展的具体贡献 目前尚不清楚，其在改变肺肿瘤代谢中的作用也不清楚。大多数（如果不是全部）胰岛素/IGF-1 信号传导 肺在细胞内会聚到接头蛋白胰岛素受体底物 IRS1 和 IRS2 上，然后 分化为下游信号传导效应器的复杂网络，包括 PI3K/Akt。叉头 转录因子 Foxo1 和 Foxo3 是胰岛素调节靶标，可被 Akt 灭活，并影响 细胞代谢、生存和增殖。 Foxos 以调节肝脏葡萄糖代谢而闻名 通过促进葡萄糖的产生并抑制其利用。然而，潜在的肿瘤抑制作用 Foxos 在 Kras 驱动的癌症中的作用尚未得到研究。在这里，使用不同的条件遗传 Kras 驱动的肺癌工程小鼠 (GEM) 模型，IRS1 和 IRS2 基因消融的影响 肺肿瘤的发生、维持和代谢，以及 Foxo1 和 Foxo3 在其中的作用 调解这些影响，将被调查。组织病理学和体内成像技术将用于 在 Kras 激活同时或之后的时间点评估 IRS 基因丢失对肺的影响 这些小鼠的肿瘤潜伏期、肿瘤负荷和存活率以及 Foxo1 和 Foxo3 基因是否额外丢失 会扭转这种影响。此外，信号通路的激活和基因表达的差异 将检测调节葡萄糖利用的物质，并对葡萄糖衍生的物质进行质谱分析 在存在或不存在 Foxo 的情况下，IRS 基因丢失后，将对肿瘤进行代谢 基因。细胞也将从 Kras 驱动的肿瘤中分离出来并在培养物中生长。类似的方法 然后将在体外对细胞进行针对体内肿瘤描述的实验，以确定肿瘤下游的靶点 IRS 和 Foxos 可以改变葡萄糖利用率，从而影响肺肿瘤的维持。这些的作用 然后可以通过敲低/过度表达研究来确认目标。此外，类似的研究还将 在已建立的 Kras 驱动的人 NSCLC 细胞系中进行，可稳定诱导 IRS1 敲低 和/或IRS2，也可以在免疫缺陷小鼠中作为皮下异种移植物生长。这些结果 研究将揭示可利用的新型代谢克拉斯驱动的肺癌漏洞 NSCLC患者的治疗。 ！