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％），分为两个主要的亚型， 肺腺癌（LADC）和肺鳞状细胞癌（LUSC）。针对性的新治疗策略 LADC的主要致癌驱动因素已提高了反应率和患者的生存率。但是，由于 缺乏特征良好的，经过验证和可热的致癌驱动因素，类似的疗法 LUSC的进步尚未实现。原代人LUSC的基因组分析表明 LUSC中最普遍的复发遗传改变是TP53的同时损失和拷贝数增长 （CNG）在3q26染色体上（> 85-90％）。在两个癌前病变中都观察到这些遗传改变 和早期的LUSC，表明它们是LUSC肿瘤发生的早期促进事件。我们以前的研究 已经证明了3Q26基因Prkci，Sox2和Ect2是协调的，并且 在功能上协作以维持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）在 TRP53损失的上下文足以转化小鼠肺碱性干细胞（LBSC），这是一个主要原点细胞 LUSC，并驱动具有恶性LUSC特征的肿瘤的形成； 2）PKCι-SOX2信号激活A 主转录程序指定谱系限制的肺鳞状转化； 3）PKCι-Ect2 信号传导功能以通过控制MEK-ERK信号传导来增加LUSC细胞的增殖潜力 并增强核糖体生物发生； 4）均具有3Q26 CNG和SOX2，PRKCI的人类LUSC细胞系 ECT2转化的LBSC对Auranofin（ANF）的生长抑制作用敏感，这是一种有效的 PKCι抑制剂，以及针对致癌PKCι-Sox2和PKCι-Ect2驱动效应子的小分子抑制剂 包括刺猬，Wnt，Notch，Mek-Erk和RRNA合成的途径。根据这些数据，我们 假设：1）PRKCI，SOX2和ECT2代表LUSC的合作多基因驱动器 肿瘤发生； 2）与PKCι-Sox2和PKCι-Ect2的ANF和抑制剂合并治疗 效应途径将协同抑制LUSC肿瘤的转化生长。这些假设会 通过完成两个相互关联的特定目的来测试：1）评估新颖的效率 靶向致癌PKCι效应途径的药物组合； 2）一般描述一个小说 - PRKCI驱动的LUSC的可处理小鼠模型。这些研究的成功完成将促进 新型治疗策略的设计以改善LUSC患者的结局。此外，我们的小说 临床上与基因工程相关的LUSC小鼠模型将增强我们对LUSC生物学的理解， 表征LUSC倡议和从肿瘤病变到恶性LUSC的发展，识别标记 用于早期LUSC诊断和开发和测试新型靶向疗法，以改善对LUSC的治疗。