Gene Regulatory Mechanisms Controlling Tissue Maturation and Polyploidization

控制组织成熟和多倍化的基因调控机制

• 批准号: 10396445
• 负责人: Auinash Kalsotra
• 金额: $ 18.78万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-22 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10396445
• 关键词: Adult; Affect; Alternative Splicing; Animals; Automobile Driving; Biochemistry; Birth; Buffers; Cell Cycle; Cell Nucleus; Cells; Chromosomes; Code; Communities; Competence; Cytokinesis; DNA Damage; DNA Sequence Alteration; Development; Diploidy; Epithelial; Event; Exhibits; Exons; Failure; Frequencies; Future; Gene Dosage; Gene Expression; Genes; Genetic; Genetic Transcription; Genome; Genomic approach; Goals; Hepatic; Hepatocarcinogenesis; Hepatocyte; Human; Injury; Knockout Mice; Link; Liver; Mammals; Metabolic; Mitosis; Molecular; Mus; Mutation; Nature; Nutritional; Organ; Outcome; Output; Oxidative Stress; Pattern; Physiological; Plants; Ploidies; Polyploid Cells; Polyploidy; Primary carcinoma of the liver cells; Production; Property; Proteins; Proteome; RNA Splicing; RNA-Binding Proteins; Regulator Genes; Research; Resolution; Resources; Rodent; Role; Spliced Genes; Testing; Time; Tissues; Transcript; Tumor Suppressor Proteins; Variant; Xenobiotics; cell type; experimental study; genetic approach; genetic regulatory protein; genome-wide; insight; liver development; loss of function; mRNA Precursor; mouse model; next generation sequencing; postnatal; postnatal period; programs; transcriptome; tumorigenesis

I. ABSTRACT Polyploidy—a state in which cells carry more than two sets of chromosomes—is frequently observed in nature, yet, the genetic mechanisms controlling ploidy and its functional significance remain enigmatic. The liver, in particular, gains a high percentage of polyploid hepatocytes during postnatal period of development; and the frequency and extent of hepatic polyploidization are further increased following injury, DNA damage, and oxidative stress, but are decreased in hepatocellular carcinoma. Recent evidence suggests that polyploidy safeguards the liver from tumorigenesis by slowing the proliferative capacity of hepatocytes and maintaining a reservoir of tumor suppressors. However, there is minimal understanding of the molecular events that govern the postnatal initiation/promotion of hepatic polyploidization or how differences in chromosomal ploidy affect the transcriptional and posttranscriptional activities of hepatocytes. We have previously demonstrated that the RNA binding protein ESRP2 is a key developmentally regulated factor, which activates an adult splicing program to facilitate terminal differentiation, functional competence, and maturation of hepatocytes. The goals of this proposal are to (i) determine the physiological necessity/sufficiency of ESRP2 and its splicing- regulatory-network in driving hepatocyte polyploidy, and (ii) define the quantitative and qualitative impact of ploidy on hepatocyte transcriptional output. Aim 1 will use ESRP2 gain-and loss-of-function mouse models to determine if programmed changes in RNA splicing through ESRP2 activation are crucial for the polyploidization of hepatocytes. In Aim 2, we will generate high-resolution transcriptomes from diploid and polyploid murine hepatocytes to investigate how ploidy influences the steady-state levels and alternative splicing patterns of hepatic transcripts at a genome-wide scale. The proposed aims will examine new gene regulatory mechanism(s) controlling polyploidization while uncovering previously unrecognized links between alternative splicing and cellular polyploidy.

I.摘要 多倍体（一种细胞携带两组以上染色体的状态）在自然界中经常观察到 然而，控制倍性的遗传机制及其功能意义仍然是神秘的。肝脏，在 特别是，在产后发育期间，多倍体肝细胞获得了很高的比例；和 损伤，DNA损伤和 氧化应激，但在肝细胞癌中得到改善。最近的证据表明多倍体 通过减慢肝细胞的增殖能力并维持A，保护肝脏免受肿瘤发生 肿瘤补充剂的水库。但是，对控制的分子事件的了解最少 产后倡议/促进肝多倍化或染色体倍性的差异如何影响 肝细胞的转录和转录后活动。我们以前已经证明了 RNA结合蛋白ESRP2是开发的关键因素，它激活成人剪接 促进末端分化，功能能力和肝细胞成熟的程序。目标 该提议的内容是（i）确定ESRP2及其剪接的必要物理/充分性 - 驱动肝细胞多倍体的调节网络，（ii）定义了定量和定性影响 肝细胞转录输出的倍性。 AIM 1将使用ESRP2增益和功能丧失鼠标模型 确定通过ESRP2激活中RNA剪接的编程变化对于 肝细胞的多倍体化。在AIM 2中，我们将从二倍体产生高分辨率转录组， 多倍体鼠肝细胞，以研究拼字质如何影响稳态水平和替代品 肝脏转录本的剪接模式在全基因组范围内。拟议的目标将检查新基因 控制多倍体化的调节机制，同时发现先前未认识的联系 替代剪接和细胞多倍体。