A novel Drosophila platform for sequential genetic manipulations in vivo

一种用于体内连续遗传操作的新型果蝇平台

• 批准号: 10193543
• 负责人: Erdem Bangi
• 金额: $ 23.1万
• 依托单位: FLORIDA STATE UNIVERSITY
• 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 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10193543
• 关键词: APC gene; Address; Adult; Animal Model; Architecture; Area; Automobile Driving; Back; Bar Codes; Biological Models; Biological Process; Cancer Model; Carcinoma; Cell Differentiation process; Cells; Cellular biology; Colonic Neoplasms; Color; Colorectal Adenoma; Colorectal Cancer; Complement; Complex; Computer Models; Coupled; Coupling; DNA Sequence Alteration; Data; Developmental Biology; Disease; Disease model; Drosophila genus; Elements; Future; Genes; Genetic; Genetic Heterogeneity; Genetic Processes; Genetic Recombination; Genome; Goals; Human; Human Characteristics; Individual; Intestines; KRAS oncogenesis; KRAS2 gene; Label; Laboratories; Life; MADH4 gene; Malignant Neoplasms; Medical; Methods; MicroRNAs; Modeling; Mutate; Mutation; Neurosciences; Normal tissue morphology; Oncogenic; Orthologous Gene; Other Genetics; Population; Proteins; Research; Resistance; Resources; Scheme; Series; Signal Pathway; Site; Solid Neoplasm; System; TP53 gene; Technology; Testing; Tissues; Transgenes; Transgenic Organisms; Treatment Failure; adenoma; anticancer research; cancer genome; cell type; clinically relevant; colon tumorigenesis; computer studies; design; flexibility; fly; genetic manipulation; in vivo; innovation; interest; knock-down; neoplastic cell; novel; overexpression; pressure; progenitor; stem; stem cell biology; stem cells; success; tool; tumor; tumorigenesis; validation studies

PROJECT SUMMARY The ability to perform sophisticated genetic manipulations and large-scale exploratory studies is a key strength of Drosophila as a model system. This proposal leverages this strength and pushes the limits of Drosophila genetics to establish a novel, innovative and ambitious platform that allows sequential introduction of genetic manipulations —each coupled with a different fluorescent protein— into individual cells. My laboratory is interested in using this platform to build and study tumors in the adult Drosophila intestine composed of genetically heterogeneous cell populations with distinct mutation profiles (i.e. subclones). We have previously used tumor sequence data as blueprints to build several colorectal cancer models by genetically manipulating Drosophila orthologs of human cancer driver genes in the adult fly intestine. We now seek to expand these efforts to building new cancer models that reflect tumor subclonal architectures identified by sequencing and computational studies. Tumors arise by sequential accumulation of genomic alterations in cancer driver genes. Consequently, most solid tumors are composed of subclones with distinct mutation profiles in constant competition for limited space and resources. Therapy often alters subclonal dynamics by introducing additional selective pressures, eventually leading to the expansion of resistant subclones and therapy failure. Building subclonal tumors is a technically challenging problem that requires sophisticated genetic manipulations and represents an important area of unmet medical need. Drosophila is a useful cancer model that captures several key hallmarks of human tumors, including colorectal cancer. The adult Drosophila intestine is a well characterized tissue with multipotent intestinal stem cells where signaling pathways altered in colon tumors are remarkably conserved and well characterized. The platform proposed here, combined with practical advantages of Drosophila, provides a unique opportunity to generate tumors with diverse subclonal architectures observed in human tumors. If successful, this technology will complement mammalian cancer models by providing an experimental platform to functionally explore yet another layer of complexity of human tumor genome landscapes revealed by big cancer data. We also hope to use this platform to explore the impact of altering the order of cancer driving genomic alterations on tumorigenesis and the potential of different cell types of the intestine as tumor cells of origin. The platform proposed here is a flexible technology that can easily be combined with other genetic tools to generate populations of cells carrying different genetic manipulations barcoded with unique fluorescent protein combinations in any tissue of interest. If successful, it could open up promising opportunities in other research areas including developmental biology, stem cell biology and neuroscience by providing a platform to explore mechanisms of cell competition and cooperation in the context of normal tissue function as well as disease.

项目摘要 进行复杂的遗传操作和大规模探索性研究的能力是关键 果蝇作为模型系统的强度。该提议利用了这种力量，并推动了 果蝇遗传学建立了一个新颖，创新和雄心勃勃的平台，该平台允许顺序引入 遗传操作 - 含有不同的荧光蛋白 - 进入单个细胞。我的实验室是 有兴趣使用此平台在成年果蝇肠中建立和研究肿瘤 具有独特的突变谱（即子克隆）的遗传异质细胞群。我们以前有 使用肿瘤序列数据作为蓝图，通过遗传操纵来构建几种大肠癌模型 成年蝇肠中人类癌驱动基因的果蝇直系同源物。我们现在寻求扩大这些努力 建立新的癌症模型，以反映通过测序和 计算研究。 肿瘤是由于癌症驱动基因中基因组改变的顺序积累而产生的。因此，大多数 实体瘤由在有限空间的恒定竞争中具有独特突变曲线的子克隆组成 和资源。治疗通常通过引入其他选择压力来改变亚克隆动力学 导致抗性亚克隆和治疗失败的扩大。从技术上讲，建筑亚克隆肿瘤是 具有挑战性的问题，需要复杂的遗传操作，并代表 未满足的医疗需求。果蝇是一种有用的癌症模型，可捕获人类肿瘤的几个关键标志， 包括结直肠癌。成年果蝇肠道是一种具有多肠的特征性组织 结肠肿瘤中信号通路改变的干细胞是明显的保守和表征的。 这里提出的平台，结合果蝇的实际优势，提供了一个独特的机会 在人类肿瘤中观察到的潜水亚克隆体系结构的肿瘤。如果成功，这项技术 将通过提供实验平台来探索哺乳动物癌症模型 大型癌症数据揭示了人类肿瘤基因组景观的另一层复杂性。我们也希望 使用此平台探索改变癌症驱动基因组改变顺序对的影响 肿瘤发生和肠道不同细胞类型的潜力作为起源的肿瘤细胞。 这里提出的平台是一种灵活的技术，可以轻松地将其与其他遗传工具结合在一起 产生携带不同遗传操作的细胞种群，该遗传操作用独特的荧光蛋白进行了条形码 任何感兴趣的组织的组合。如果成功，它可能会在其他研究中为承诺机会开放 通过提供探索平台，包括发育生物学，干细胞生物学和神经科学在内的领域 在正常组织功能和疾病的背景下，细胞竞争与合作的机制。