Modeling the KIF5B-RET Fusion Gene of Lung Cancer in Vivo

体内肺癌 KIF5B-RET 融合基因建模

• 批准号: 8617823
• 负责人: JIE WU
• 金额: $ 21.33万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2015-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8617823
• 关键词: Adenocarcinoma; Animal Model; Animals; Apoptosis; Cancer Etiology; Cancer Patient; Cessation of life; Chemotherapy-Oncologic Procedure; Clinical; Communities; Development; Disease; Doxycycline; Drug Targeting; Effectiveness; Epidermal Growth Factor Receptor; Epithelial; Evaluation; Family member; Fibroblasts; Gene Fusion; Generations; Genes; Genetic; Genetic Engineering; Genetically Engineered Mouse; Genomics; Goals; Human; Hyperplasia; Individual; Kinesin; Lesion; Lung; Lung Adenocarcinoma; Lung Neoplasms; Maintenance; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Methods; Microinjections; Modeling; Molecular; Mus; Mutant Strains Mice; Mutation; Non-Small-Cell Lung Carcinoma; Oncogenes; Oncogenic; PTPN11 gene; Patients; Pharmaceutical Preparations; Phenocopy; Phenotype; Plasmids; Pre-B Lymphocyte; Pre-Clinical Model; Protein Tyrosine Kinase; RET gene; Recurrence; Research; Resistance; Resources; Role; Site; Techniques; Technology; Tetracyclines; Time; Trans-Activators; Transgenes; Transgenic Mice; Transgenic Organisms; Translating; Xenograft procedure; alveolar type II cell; base; chemotherapy; clinically relevant; comparative; cytotoxic; fusion gene; human disease; improved; in vivo; inhibitor/antagonist; lung development; mouse Cre recombinase; mouse model; mutant; neoplastic; neoplastic cell; novel; offspring; oncogene addiction; pre-clinical; preclinical evaluation; preclinical study; public health relevance; recombinase; response; transgene expression; tumor; zygote

DESCRIPTION (provided by applicant): Lung cancer is a leading cause of cancer death in the world. Conventional cytotoxic-based chemotherapy regimens are at their limits of maximal effectiveness. New treatment paradigms incorporating inhibitors to specific driver mutations, such as EGFR and ALK, have shown improved clinical benefits. These new treatments depend on identification and evaluation of clinically relevant genetic lesions that drive and maintain the malignancy. Lung cancer is a heterogeneous disease. Clinically relevant mutations in ~50% of non-small cell lung cancer (NSCLC) remain to be identified and evaluated. It is envisioned that recent advances in genomic technologies will accelerate the discovery of new genetic alternations in human cancer. An important aspect in translating the new genomic findings into improved treatments is to have in vivo preclinical models that faithfully phenocopy the newly identified genes in the human diseases for evaluating drug response and resistance. Recurrent gene fusion between kinesin family member 5B (KIF5B) and the RET tyrosine kinase (KIF5B-RET) is the most recent genetic alternation identified in human lung adenocarcinoma. Preclinical models of KIF5B-RET have been limited to xenografts of transfected NIH3T3 fibroblasts and Ba/F3 pre-B- lymphocytes. The goal of this study is to generate doxycycline-inducible, lung-specific KIF5B-RET transgenic mice by Cre recombinase-mediated cassette exchange (Cre-RMCE) to model the oncogenic activity of KIF5B- RET in lung type II alveolar cells in bitransgenic mice. In Specific Aim 1, plasmids carrying a tetO-KIF5B-RET cassette flanked by heterospecific L2/L3 loxP sites and Cre will be co-injected into fertilized eggs (zygotes) derived from different lines of existing Cre-RMCE-capable transgenic mice to replace the original floxed tetO- SHP2 transgenes with tetO-KIF5B-RET. In Specific Aim II, inducible KIF5B-RET expression and effects of KIF5B-RET on lung tumor development and maintenance will be evaluated in CCSP-rtTA/tetO-KIF5B-RET bitransgenic mice. Since our Cre-RMCE-capable SHP2 transgenic mice have already been characterized for inducible expression and function of the transgene in the lung, new transgenic mice derived from these lines by cassette exchange are expected to display reliable inducible transgene expression. This will accelerate the generation and examination of novel useful transgenic mouse models. Thus, the study will generate genetically engineered animals urgently needed for development of new therapy targeting the KIF5B-RET molecular subtype of lung adenocarcinoma. Furthermore, the study will identify the most efficient Cre-RMCE- capable transgenic line(s) harboring a single copy of the KIF5B-RET transgene for subsequent studies to derive new transgenic mice. This will provide an important resource for the research community to accelerate the generation of mouse models of newly identified molecular lesions.

描述（由申请人提供）：肺癌是世界上癌症死亡的主要原因。传统的基于细胞毒的化疗方案已达到最大有效性的极限。将抑制剂结合到特定驱动突变（例如 EGFR 和 ALK）的新治疗范例已显示出改善的临床益处。这些新疗法取决于对临床相关遗传病变的识别和评估，这些病变驱动和维持 恶性肿瘤。肺癌是一种异质性疾病。约 50% 的非小细胞肺癌 (NSCLC) 中的临床相关突变仍有待识别和评估。预计基因组技术的最新进展将加速人类癌症中新基因变异的发现。将新的基因组发现转化为改进的治疗方法的一个重要方面是建立体内临床前模型，忠实地复制人类疾病中新发现的基因，以评估药物反应和耐药性。驱动蛋白家族成员 5B (KIF5B) 和 RET 酪氨酸激酶 (KIF5B-RET) 之间的反复基因融合是人肺腺癌中最新发现的基因变异。 KIF5B-RET 的临床前模型仅限于转染的 NIH3T3 成纤维细胞和 Ba/F3 前 B 淋巴细胞的异种移植物。本研究的目标是通过 Cre 重组酶介导的盒交换 (Cre-RMCE) 产生强力霉素诱导的肺特异性 KIF5B-RET 转基因小鼠，以模拟双转基因小鼠肺 II 型肺泡细胞中 KIF5B-RET 的致癌活性。在具体目标 1 中，携带侧翼为异特异性 L2/L3 loxP 位点和 Cre 的 tetO-KIF5B-RET 盒的质粒将被共注射到来自现有 Cre-RMCE 能力转基因小鼠的不同品系的受精卵（受精卵）中，以用 tetO-KIF5B-RET 取代原来的 floxed tetO-SHP2 转基因。在Specific Aim II中，将在CCSP-rtTA/tetO-KIF5B-RET双转基因小鼠中评估诱导型KIF5B-RET表达以及KIF5B-RET对肺肿瘤发展和维持的影响。由于我们的具有 Cre-RMCE 能力的 SHP2 转基因小鼠已经被表征为转基因在肺中的诱导表达和功能，因此通过盒交换从这些品系衍生的新转基因小鼠预计将表现出可靠的诱导转基因表达。这将加速新型有用的转基因小鼠模型的产生和检查。因此，该研究将产生开发针对肺腺癌 KIF5B-RET 分子亚型的新疗法迫切需要的基因工程动物。此外，该研究将确定最有效的具有 Cre-RMCE 能力的转基因品系，该品系含有单个 KIF5B-RET 转基因拷贝，用于后续研究以衍生新的转基因小鼠。这将为研究界加速生成新发现的分子病变的小鼠模型提供重要资源。