Drosophila models of Ret fusions in Papillary Thyroid Carcinoma

甲状腺乳头状癌 Ret 融合果蝇模型

• 批准号: 8784857
• 负责人: Sarah Levinson
• 金额: $ 3.53万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8784857
• 关键词: Accounting; Affect; Apoptosis; Automobile Driving; Benign; Biochemical; Bioinformatics; Biological Assay; CCDC6 gene; Cancer cell line; Cell Line; Cells; Cessation of life; Chimeric Proteins; Clinical Trials; Coiled-Coil Domain; Collaborations; Cytoplasmic Protein; Data; Disease; Dominant Genetic Conditions; Drosophila genus; Ensure; Gene Expression Profile; Gene Mutation; Genes; Genetic; Genetic screening method; Goals; Human; In Vitro; Individual; Indolent; Knowledge; Larva; Lead; Libraries; Lung Adenocarcinoma; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of thyroid; Modeling; Monitor; Mutation; Oncogenic; PTCH gene; Papillary thyroid carcinoma; Pathway Analysis; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Phosphotransferases; Protein Isoforms; RNA Interference; Receptor Protein-Tyrosine Kinases; Refractory Disease; Research Design; Role; Sampling; Signal Pathway; Signal Transduction; Solid; Survival Rate; Testing; The Cancer Genome Atlas; Therapeutic; Thyroid Gland; Transgenes; Variant; Western Blotting; Work; base; cell growth; clinically relevant; clinically significant; effective therapy; feeding; fly; in vivo; inhibitor/antagonist; innovation; insight; kinase inhibitor; medullary thyroid carcinoma; migration; outcome forecast; promoter; public health relevance; response; screening; senescence; success; therapy development; transcriptome sequencing; tumor

DESCRIPTION (provided by applicant): Rates of thyroid cancer have tripled in the last few decades, making it the fastest growing form of cancer. 80% of thyroid cancer cases are Papillary Thyroid Carcinoma (PTC), which arises from transformation of follicular cells in the thyroid. Metastatic PTC has no effective treatment, a five-year survival rate of 50%, and results in the death of approximately 1500 patients in the US each year. 5-30% of PTC cases involve fusion proteins in which the Ret receptor tyrosine kinase is fused to the coiled-coiled domain of various cytoplasmic proteins. Recently, Ret fusions have also been identified in a subset of lung adenocarcinomas. Thus, therapeutics for Ret fusion-driven tumors remains an unmet need. Two Ret fusions, CCDC6-RET (PTC1) and NcoA4-RET (PTC3) account for greater than 90% of fusions found in PTC. Although both fusion proteins lead to activation of Ret, the two fusions are associated with different PTC subtypes. CCDC6-RET is closely associated with the classic variant, a more benign subtype, and NcoA4-RET is closely associated with the solid subtype, which is more aggressive and malignant. Work by the Cagan lab has validated Drosophila models of oncogenic Ret isoforms that are associated with Medullary Thyroid Carcinoma (MTC). These models were used to explore function as well as identify lead therapeutic hits. The aim of this proposal is to characterize the signaling pathways activated by Ret fusions CCDC6-RET and NcoA4-RET, identify differences that are clinically significant, and develop an optimized therapy. First, signaling changes caused by expression of CCDC6-RET and NcoA4-RET in Drosophila will be characterized and compared by Western blot analysis and immunoflourescence. Secondly, clinically relevant kinase inhibitors will be fed to flies expressing either Ret fusion to identify potential therapies based on rescue of transgene-induced phenotypes. Lastly, the relevance of genetic complexity of PTC tumors on treatment will be evaluated using a functional driver vs. passenger screen using a pre-existing Drosophila kinome RNAi library. All identified therapies will be evaluated in mammalian models, such as cancer cell lines, to ensure the findings are relevant to patients. The innovative studies proposed here will provide critical insight into how Ret fusions drive PTC, how different Ret fusions may differ in their effects, and how Ret fusion driven tumors may respond to targeted therapies.

描述（由申请人提供）：甲状腺癌的发病率在过去几十年中增加了两倍，使其成为增长最快的癌症形式。 80％的甲状腺癌病例是甲状腺乳头状癌（PTC），由甲状腺滤泡细胞的转化引起。转移性PTC尚无有效治疗方法，五年生存率为50%，在美国每年导致约1500名患者死亡。 5-30% 的 PTC 病例涉及融合蛋白，其中 Ret 受体酪氨酸激酶与各种细胞质蛋白的卷曲螺旋结构域融合。最近，Ret 融合也在肺腺癌中被发现。因此，Ret 融合驱动的肿瘤的治疗仍然是一个未得到满足的需求。 CCDC6-RET (PTC1) 和 NcoA4-RET (PTC3) 这两种 Ret 融合占 PTC 中发现的融合的 90% 以上。尽管两种融合蛋白都会导致 Ret 激活，但这两种融合蛋白与不同的 PTC 亚型相关。 CCDC6-RET 与经典变异（一种更良性的亚型）密切相关，NcoA4-RET 与实体亚型（更具侵袭性和恶性）密切相关。 Cagan 实验室的工作验证了与甲状腺髓样癌 (MTC) 相关的致癌 Ret 亚型的果蝇模型。这些模型用于探索功能并确定主要治疗效果。该提案的目的是表征 Ret 融合 CCDC6-RET 和 NcoA4-RET 激活的信号通路，识别具有临床意义的差异，并开发优化的治疗方法。首先，将通过蛋白质印迹分析和免疫荧光来表征和比较果蝇中 CCDC6-RET 和 NcoA4-RET 表达引起的信号变化。其次，将临床相关激酶抑制剂喂给表达表达的果蝇 要么通过 Ret 融合来识别基于拯救转基因诱导表型的潜在疗法。最后，将使用预先存在的果蝇激酶组 RNAi 库，通过功能驱动与乘客筛选来评估 PTC 肿瘤的遗传复杂性与治疗的相关性。所有已确定的疗法都将在哺乳动物模型（例如癌细胞系）中进行评估，以确保研究结果与患者相关。这里提出的创新研究将为 Ret 融合如何驱动 PTC、不同 Ret 融合的效果有何不同，以及 Ret 融合驱动的肿瘤如何对靶向治疗做出反应提供重要见解。