Control of cell proliferation and differentiation by the Retinoblastoma tumor suppressor pathway

视网膜母细胞瘤肿瘤抑制途径控制细胞增殖和分化

• 批准号: 9908112
• 负责人: Maxim Frolov
• 金额: $ 57.02万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9908112
• 关键词: Address; Adult; Affect; Animal Model; Animals; Apoptosis; Atlases; Cell Differentiation process; Cell Proliferation; Cells; Complement; Development; Drosophila genus; E2F transcription factors; Event; Eye; Family; Fat Body; Gene Expression; Genetic; Genomics; Growth; Human; Investigation; Knowledge; Link; Malignant Neoplasms; Metabolism; Muscle Development; Mutation; Myofibrillogenesis; Pathway interactions; Photoreceptors; Population; Research; Retinoblastoma; Retinoblastoma Protein; Skeletal Muscle; System; Technology; Tissues; Tumor Suppressor Proteins; cell type; droplet sequencing; fly; genetic analysis; imaginal disc; in vivo; innovation; member; mutant; programs; retinoblastoma tumor suppressor; single-cell RNA sequencing

PROJECT SUMMARY/ABSTRACT Inactivation of the retinoblastoma (Rb) tumor suppressor pathway is an early obligatory event in human cancer. One of the key targets of the Rb pathway is the family of E2F transcription factors, which regulate cell proliferation and apoptosis. Overlapping and redundant functions of mammalian members of the E2F and Rb tumor suppressor protein (pRB) families is a major hurdle in identification of their in vivo function. Model organisms such as Drosophila provide attractive alternatives to complement studies in mammalian systems due to high conservation of the Rb pathway and the amenability of flies to genetic analysis. Our research program is aimed at addressing two fundamental questions: what are the essential functions of the Rb pathway in development, and how do Rb pathway mutations affect different cell types in the tissue? To address these questions, we are using genetic, genomic and single-cell RNA-Seq approaches. We have identified adult skeletal muscle where E2F and Rb proteins regulate growth and myofibrillogenesis, to be important for viability. We will determine how the Rb pathway regulates gene expression and cellular metabolism during muscle development, and will investigate the systemic effects inferred by E2F deficiency in specific tissues such as adult skeletal muscle and the fat body. We have adapted Drop-Seq, a single-cell RNA-sequencing technology, to determine the effect of Rb pathway mutations in heterogeneous tissue. We have established the feasibility of this approach by building the first cell atlas of the Drosophila wild-type eye imaginal disc. Profiling of Rb mutants by Drop-Seq revealed elevated intracellular acidification in a small cell population that makes them uniquely sensitive to apoptosis. The link between apoptosis and acidification will be evaluated further. Among other questions to be addressed with Drop-seq is the investigation of why inactivation of the Hippo pathway induces dedifferentiation of Rb mutant photoreceptors. Collectively, the proposed research will enhance our knowledge of pRB and its functions during animal development.

项目摘要/摘要 视网膜母细胞瘤（RB）肿瘤抑制途径的失活是人类的早期强制性事件 癌症。 RB途径的关键目标之一是E2F转录因子的家族，该系数调节 细胞增殖和凋亡。 E2F的哺乳动物成员的重叠和冗余功能 RB肿瘤抑制蛋白（PRB）家族是鉴定其体内功能的主要障碍。 果蝇等模型生物为哺乳动物的补充研究提供了有吸引力的替代品 由于对RB途径的高保守性以及果蝇对遗传分析的不及格而导致的系统。我们的 研究计划旨在解决两个基本问题： RB发育中的途径以及RB途径突变如何影响不同的细胞类型 组织？为了解决这些问题，我们正在使用遗传，基因组和单细胞RNA-seq方法。 我们已经确定了成年骨骼肌，其中E2F和RB蛋白调节生长和 肌原纤维发生，对于生存能力很重要。我们将确定RB途径如何调节基因 肌肉发育过程中的表达和细胞代谢，并将研究系统性作用 由E2F缺乏在特定组织（例如成人骨骼肌和脂肪体）中推断出来。我们有 改编的Drop-Seq是一种单细胞RNA测序技术，以确定RB途径的效果 异质组织中的突变。我们通过建立 果蝇野生型眼盘的第一个细胞图集。通过Drop-seq揭示了RB突变体的分析 小细胞群中的细胞内酸化升高，这使它们对 凋亡。凋亡与酸化之间的联系将进一步评估。除其他问题 drop-seq解决的是研究为什么河马途径失活的研究 RB突变体光感受器的去分化。拟议的研究共同增强了我们的 对PRB及其在动物发展过程中的功能的了解。