Protein kinase C and lung carcinogenesis

蛋白激酶C与肺癌发生

• 批准号: 9126982
• 负责人: MARCELO G. KAZANIETZ
• 金额: $ 39.12万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9126982
• 关键词: A/J Mouse; Ablation; Address; Air Pollutants; Alleles; Aromatic Polycyclic Hydrocarbons; Benzo(a)pyrene; Bioinformatics; Breast; Cancer Burden; Cancer Etiology; Carcinogens; Case Study; Cell model; Cells; Cessation of life; Chemical Actions; Codon Nucleotides; Computer Simulation; DNA; Data; Databases; Development; Disease; Elements; Environmental Carcinogens; Epigenetic Process; Epithelial; Epithelial Cells; Etiology; Event; Exposure to; Frequencies; Genes; Genetic; Goals; Growth; Head and Neck Cancer; Health; Human; Induced Mutation; KRAS2 gene; Knockout Mice; Laboratories; Lead; Lesion; Link; Longevity; Lung; Lung Adenocarcinoma; Lung Neoplasms; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Mediator of activation protein; Mesenchymal; Modeling; Molecular; Mus; Mutate; Mutation; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Nude Mice; Oncogenic; Patients; Phenotype; Phosphotransferases; Play; Premalignant; Process; Prostate; Protein Kinase C; RNA Interference; Resistance; Risk; Role; Signal Transduction; Solid; Stem cells; Testing; Therapeutic; Transgenic Mice; Up-Regulation; Urethane; autocrine; base; cancer cell; cancer initiation; cancer therapy; carcinogenesis; cell motility; chemical carcinogen; environmental chemical; expectation; gain of function; genetic signature; in vivo; inducible gene expression; inhibitor/antagonist; insight; lung carcinogenesis; lung tumorigenesis; member; mouse model; mutant; neoplastic; novel; overexpression; prevent; protein kinase C epsilon; prototype; public health relevance; response; translational medicine; tumor; tumor progression; tumorigenesis; tumorigenic

 DESCRIPTION (provided by applicant): This application focuses on the study of novel mechanisms involved in early events of lung carcinogenesis. Environmental carcinogens are major causative agents of lung cancer, as they induce genetic and epigenetic alterations that ultimately lead to the malignant transformation of lung epithelial cells. Oncogenic mutations in KRAS, a common alteration in non-small cell lung cancer (NSCLC), are induced in high frequency by lung carcinogens such as polycyclic aromatic hydrocarbons (PAHs) and many other environmental carcinogens. It has been established that protein kinase C epsilon (PKC), a mitogenic, pro-survival, and tumorigenic kinase, is up-regulated in epithelial cancers, including NSCLC. Studies from our laboratory revealed that PKC is an essential mediator of tumor formation, invasiveness, and metastasis of NSCLC cells. More recently, we developed a mouse model for inducible lung-specific expression of KRas in a PKC-deficient background (LSL- K-rasG12D; PKC-/-), and found that genetic ablation of the PKC gene (PRKCE) markedly impairs the formation of tumors driven by the activated KRas allele. This suggests that PKC is required for the initiation of KRas lung tumorigenesis. Moreover, in silico database analysis in KRAS mutated human lung adenocarcinomas revealed a significant association between high PKC expression and short overall patient survival. Altogether, this led us to hypothesize that PKC is a necessary mediator of the actions of lung carcinogens. In Specific Aim 1 we will examine if PKC KO mice (in A/J genetic background) are resistant to the effects of lung carcinogens known to induce mutations in KRas, including the PAH benzo[a]pyrene (B[a]P) and urethane. We will examine if a pharmacological inhibitor of PKC (V1-2) inhibits the formation of lung tumors induced by these carcinogens or by an activated KRas allele. Mechanistic studies will be pursued to assess if carcinogens up-regulate PKCin lung epithelial cells. In Specific Aim 2, we will use genetic and pharmacological approaches to determine if PKC mediates the expansion of lung cancer progenitor cells (bronchioalveolar stem cells or BASCs) required for KRas- and carcinogen-induced tumorigenesis. To unambiguously establish a role for PKC in initiation we will use gain-of-function approaches in cellular models as well as generate an inducible lung-specific transgenic mouse line for this kinase to determine if this leads to the formation of pre-malignant or malignant lung lesions. Finally, in Specific Aim 3 we will dissect the mechanistic basis for the functional interaction between KRas and PKC in lung cancer, focusing on a) the analysis of elements of the Ras cascade, b) the identification of a PKC gene signature and transcriptional networks regulated by this kinase in the context of KRas tumorigenesis, and c) the assessment of a potential role for PKC in epithelial-mesenchymal transition (EMT), a process required for the acquisition of invasive capacity of lung cancer cells. Our studies should provide novel mechanistic insights into the molecular effects of environmental carcinogens as well as reveal important aspects of early events of lung carcinogenesis, thus impacting on our understanding of lung cancer etiology.

 描述（由申请人提供）：本申请重点研究涉及肺癌早期事件的新机制。环境致癌物是肺癌的主要致病因素，因为它们会诱导遗传和表观遗传改变，最终导致肺上皮细胞恶性转化。 KRAS 的致癌突变是非小细胞肺癌 (NSCLC) 的常见改变，由多环芳烃 (PAH) 等肺癌致癌物和许多其他环境致癌物高频诱导。已确定蛋白激酶 C epsilon (PKC) 是一种促有丝分裂、促生存和致瘤激酶，在上皮癌中表达上调，包括 非小细胞肺癌。我们实验室的研究表明，PKC是NSCLC细胞肿瘤形成、侵袭和转移的重要介质。最近，我们开发了一种在 PKC 缺陷背景下诱导肺特异性 KRas 表达的小鼠模型（LSL- K-rasG12D；PKC-/-），并发现 PKC 基因 (PRKCE) 的基因消除显着损害由激活的 KRas 等位基因驱动的肿瘤的形成。这表明 PKC 是 KRas 肺部肿瘤发生的启动所必需的。此外，KRAS 突变人肺腺癌的计算机数据库分析揭示了高 PKC 表达与患者总生存期短之间存在显着相关性。总而言之，这使我们推测 PKC 是肺癌致癌物作用的必要介质。在具体目标 1 中，我们将检查 PKC KO 小鼠（A/J 遗传背景）是否对已知可诱导 KRas 突变的肺癌致癌物（包括多环芳烃苯并[a]芘 (B[a]P) 和氨基甲酸酯）的影响具有抵抗力。我们将检查 PKC (V1-2) 的药理学抑制剂是否抑制形成 由这些致癌物或激活的 KRas 等位基因诱发的肺部肿瘤。将进行机制研究以评估致癌物是否上调肺上皮细胞中的 PKC。在具体目标 2 中，我们将使用遗传和药理学方法来确定 PKC 是否介导 KRas 和致癌物诱导的肿瘤发生所需的肺癌祖细胞（支气管肺泡干细胞或 BASC）的扩增。为了明确确定 PKC 在起始过程中的作用，我们将在细胞模型中使用功能获得方法，并为该激酶生成可诱导的肺特异性转基因小鼠系，以确定这是否会导致恶性前或恶性肺部病变的形成。最后，在具体目标 3 中，我们将剖析肺癌中 KRas 和 PKC 之间功能相互作用的机制基础，重点是 a) Ras 级联元素的分析，b) 在 KRas 肿瘤发生背景下鉴定 PKC 基因特征和受该激酶调节的转录网络，以及 c) 评估 PKC 在上皮间质转化中的潜在作用 (EMT)，肺癌细胞获得侵袭能力所需的过程。我们的研究应该为环境致癌物的分子效应提供新的机制见解，并揭示肺癌早期事件的重要方面，从而影响我们对肺癌病因学的理解。