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: 9895645
• 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/9895645
• 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; 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. !

项目总结/摘要 非小细胞肺癌（NSCLC）占肺癌的大多数，迄今为止仍然是肺癌的主要类型。 在美国和全世界癌症死亡的主要原因。约25%的NSCLC携带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在研究中。 研究和替代治疗方法是迫切需要的。药理学证据和 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 肺癌然而，该途径对Kras驱动的肿瘤起始和进展的具体贡献 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，如果不是所有的胰岛素/IGF-1信号传导， 肺在细胞内会聚到衔接蛋白胰岛素受体底物IRS 1和IRS 2上， 发散到下游信号效应物的复杂网络，包括PI 3 K/Akt。叉头 转录因子Foxo 1和Foxo 3是胰岛素调节的靶点，它们被Akt灭活， 细胞代谢、存活和增殖。众所周知，Foxos可以调节肝脏葡萄糖代谢 通过促进葡萄糖的产生和抑制其利用。然而，潜在的肿瘤抑制 Foxos在Kras驱动的癌症中的作用尚未研究。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，IRS 1和IRS 2基因消融的影响 对肺肿瘤的发生、维持和代谢的作用，以及Foxo 1和Foxo 3在 将对这些影响进行研究。组织学和体内成像技术将用于 在Kras激活的同时或之后的时间点评估IRS基因缺失对肺的影响， 这些小鼠的肿瘤潜伏期、肿瘤负荷和生存率以及Foxo 1和Foxo 3基因是否额外丢失 将扭转这种影响。此外，信号通路和基因表达的差异激活 将测定调节葡萄糖利用的蛋白质，并对葡萄糖衍生的蛋白质进行质谱分析。 在存在或不存在Foxo的情况下，当IRS基因丢失时， 基因.细胞也将从Kras驱动的肿瘤中分离并在培养物中生长。方法类似于 然后，将对体外细胞进行针对体内肿瘤所描述的那些，以鉴定下游靶点。 IRS和Foxos，可以改变葡萄糖利用，从而影响肺肿瘤的维持。这些的作用 然后可以通过敲低/过表达研究来确认靶。此外，类似的研究将在 在已建立的Kras驱动的人NSCLC细胞系中进行，具有稳定的诱导型IRS 1敲低 和/或IRS 2，其也可以作为皮下异种移植物在免疫缺陷小鼠中生长。从这些 研究将揭示新的代谢Kras驱动的肺癌漏洞，可以利用 治疗NSCLC患者。 !