A Genetically Tractable Mouse Model for PRKCI-driven Lung Squamous Cell Carcinoma

PRKCI 驱动的肺鳞状细胞癌的遗传易控制小鼠模型

• 批准号: 10296271
• 负责人: Alan P. Fields
• 金额: $ 35.8万
• 依托单位: MAYO CLINIC JACKSONVILLE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10296271
• 关键词: 3q26; Alleles; Auranofin; Biochemical; Biogenesis; Bioinformatics; Biological Markers; Biology; Cancer Etiology; Cell Line; Cells; Cessation of life; Characteristics; Chromosomes; Clinical; Combined Modality Therapy; Data; Development; Diagnosis; Drug Combinations; Enterobacteria phage P1 Cre recombinase; Erinaceidae; Event; Exhibits; Genes; Genetic; Genetic Engineering; Genetic Recombination; Genetic Transcription; Genetically Engineered Mouse; Genomics; Growth; Histologic; Human; In Vitro; Keratin; Knock-in; Knowledge; Lesion; Lung; Lung Adenocarcinoma; Lung Neoplasms; MEKs; Malignant - descriptor; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of lung; Modeling; Molecular; Morphology; Mus; Mutation; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Oncogenic; Outcome; Pathway interactions; Patient-Focused Outcomes; Patients; Pharmacology; Phenotype; Prognosis; Proteins; Recombinant DNA; Recurrence; Resources; Ribosomal RNA; Ribosomes; Signal Transduction; Specific qualifier value; Squamous Cell Lung Carcinoma; Squamous cell carcinoma; TP53 gene; Testing; Therapeutic; Transgenes; United States; World Health Organization; base; cancer diagnosis; cell growth; cell transformation; clinically relevant; conditional knockout; design; druggable target; early detection biomarkers; efficacy evaluation; improved; improved outcome; in vivo; inhibitor/antagonist; mouse model; neoplastic cell; new therapeutic target; notch protein; novel; novel drug combination; novel therapeutic intervention; novel therapeutics; overexpression; pharmacodynamic biomarker; pre-clinical; predictive marker; premalignant; programs; promoter; recombinant adenovirus; response; self-renewal; small molecule inhibitor; stem cells; tumor; tumor growth; tumor initiation; tumor progression; tumorigenesis

Project Abstract Lung cancer is the number one cause of cancer death in the United States. Non-small cell lung cancer (NSCLC), which accounts for the majority (80%) of lung cancer diagnoses, is divided into two major sub-types, lung adenocarcinoma (LADC) and lung squamous cell carcinoma (LUSC). New therapeutic strategies targeting major oncogenic drivers of LADC have led to improved response rates and patient survival. However, due to a lack of well-characterized, validated, and therapeutically-actionable oncogenic drivers, similar therapeutic advances for LUSC have not been forthcoming. Genomic analysis of primary human LUSC reveals that the most prevalent recurrent genetic alterations in LUSC are concurrent loss of TP53 and copy number gains (CNGs) at chromosome 3q26 (>85-90%). These genetic alterations are observed in both precancerous lesions and early-stage LUSC, indicating they are early promotive events in LUSC tumorigenesis. Our previous studies have demonstrated that the 3q26 genes, PRKCI, SOX2 and ECT2 are coordinately co-amplified and functionally collaborate to maintain the transformed phenotype of LUSC cells. PKCι, the protein product for PRKCI, directly phosphorylates and regulates the oncogenic function of SOX2 and ECT2 in LUSC cells. Furthermore, our preliminary data demonstrate that: 1) overexpression of SOX2, PRKCI and ECT2, in the context of Trp53 loss, is sufficient to transform mouse lung basal stem cells (LBSCs), a major cell of origin for LUSC, and drive formation of tumors with malignant LUSC characteristics; 2) PKCι-SOX2 signaling activates a master transcriptional program specifying lineage-restricted lung squamous transformation; 3) PKCι-ECT2 signaling functions to increase the proliferative potential of LUSC cells through control of MEK-ERK signaling and enhanced ribosome biogenesis; and 4) both human LUSC cell lines with 3q26 CNGs and SOX2, PRKCI and ECT2 transformed LBSCs are sensitive to the growth inhibitory effects of Auranofin (ANF), a potent PKCι inhibitor, and to small molecule inhibitors of oncogenic PKCι-SOX2 and PKCι-ECT2 driven effector pathways including Hedgehog, WNT, NOTCH, MEK-ERK and rRNA synthesis. Based on these data we hypothesize that: 1) PRKCI, SOX2 and ECT2 represent cooperative multigenic drivers of LUSC tumorigenesis; and 2) combined treatment with ANF and inhibitors of PKCι-SOX2 and PKCι-ECT2 effector pathways will synergistically inhibit transformed growth of LUSC tumors. These hypotheses will be tested through completion of two interrelated specific aims designed to: 1) evaluate the efficacy of novel drug combinations that target oncogenic PKCι effector pathways; and 2) characterize a novel genetically- tractable mouse model for PRKCI-driven LUSC. Successful completion of these studies will facilitate the design of novel therapeutic strategies to improve outcomes for patients with LUSC. Furthermore, our novel clinically-relevant genetically-engineered LUSC mouse model will enhance our understanding of LUSC biology, characterize LUSC initiation and progression from preneoplastic lesions to malignant LUSC, identify markers for early LUSC diagnosis and develop and test novel targeted therapies for improved treatment of LUSC.

项目摘要 肺癌是美国癌症死亡的第一大原因。非小细胞肺癌 (NSCLC) 占肺癌诊断的大部分 (80%)，分为两个主要亚型， 肺腺癌（LADC）和肺鳞状细胞癌（LUSC）。新的治疗策略针对 LADC 的主要致癌驱动因素提高了缓解率和患者生存率。然而，由于一个 缺乏特征明确、经过验证且可治疗的致癌驱动因素、类似的治疗方法 LUSC 的预付款尚未到位。原代人类 LUSC 的基因组分析表明 LUSC 中最常见的复发性遗传改变是同时发生 TP53 丢失和拷贝数增加 (CNG) 位于染色体 3q26 (>85-90%)。这些基因改变在两种癌前病变中均观察到 和早期 LUSC，表明它们是 LUSC 肿瘤发生的早期促进事件。我们之前的研究 已经证明 3q26 基因 PRKCI、SOX2 和 ECT2 是协同共扩增的 功能上协作以维持 LUSC 细胞的转化表型。 PKCι，蛋白质产品 PRKCI 直接磷酸化并调节 LUSC 细胞中 SOX2 和 ECT2 的致癌功能。 此外，我们的初步数据表明：1）SOX2、PRKCI 和 ECT2 的过度表达，在 Trp53 缺失的情况下，足以转化小鼠肺基底干细胞 (LBSC)，这是 LUSC，并驱动具有恶性LUSC特征的肿瘤的形成； 2) PKCι-SOX2 信号传导激活 主转录程序指定谱系限制性肺鳞状细胞转化； 3) PKCι-ECT2 通过控制 MEK-ERK 信号传导增强 LUSC 细胞的增殖潜力 和增强核糖体生物合成； 4) 具有 3q26 CNG 和 SOX2、PRKCI 的人类 LUSC 细胞系 ECT2 转化的 LBSC 对金诺芬 (ANF) 的生长抑制作用敏感，金诺芬 (ANF) 是一种有效的药物 PKCι 抑制剂，以及致癌 PKCι-SOX2 和 PKCι-ECT2 驱动效应子的小分子抑制剂 途径包括 Hedgehog、WNT、NOTCH、MEK-ERK 和 rRNA 合成。根据这些数据我们 假设：1) PRKCI、SOX2 和 ECT2 代表 LUSC 的合作多基因驱动因素 肿瘤发生； 2) ANF与PKCι-SOX2和PKCι-ECT2抑制剂的联合治疗 效应途径将协同抑制 LUSC 肿瘤的转化生长。这些假设将 通过完成两个相互关联的具体目标进行测试，旨在：1）评估新颖的功效 针对致癌 PKCι 效应通路的药物组合； 2）描述一种新颖的基因特征—— PRKCI 驱动的 LUSC 的易驯化小鼠模型。成功完成这些研究将有助于 设计新的治疗策略以改善 LUSC 患者的预后。此外，我们的小说 临床相关的基因工程 LUSC 小鼠模型将增强我们对 LUSC 生物学的理解， 描述 LUSC 的起始和从癌前病变到恶性 LUSC 的进展，识别标记物 用于早期 LUSC 诊断，并开发和测试新的靶向疗法以改善 LUSC 的治疗。