Transcriptional Factor SOX2, LncRNA HBL1, microRNA1 and PRC2 Epigenetic Complex Compose a Network to Orchestrate Cardiac Differentiation from Human Pluripotent Stem Cells

转录因子 SOX2、LncRNA HBL1、microRNA1 和 PRC2 表观遗传复合物组成一个网络来协调人类多能干细胞的心脏分化

• 批准号: 9796511
• 负责人: Lei Yang
• 金额: $ 50.2万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9796511
• 关键词: Binding; CRISPR/Cas technology; Cardiac; Cardiac Myocytes; Cardiac development; Cardiovascular system; Cell Nucleus; Cells; ChIP-seq; Complex; Congenital Heart Defects; Cytoplasm; Development; Drosophila genus; Electrophysiology (science); Epigenetic Process; Exhibits; Gene Expression; Genes; Genetic; Genetic Transcription; Heart; Heterogeneity; Human; Knock-out; Laboratories; Mediating; Mesoderm; Messenger RNA; MicroRNAs; Modification; Molecular; Morphogenesis; Mus; Names; Nuclear; Organogenesis; Outcome; Pathogenicity; Pathway interactions; Play; Pluripotent Stem Cells; Polycomb; Property; Proteins; Recording of previous events; Regulator Genes; Role; Signal Transduction; Stem Cell Research; Stretching; Testing; Tissues; Transcript; Untranslated RNA; Xenopus; cardiogenesis; cell fate specification; histone modification; human pluripotent stem cell; human stem cells; induced pluripotent stem cell; knock-down; novel; overexpression; pluripotency; progenitor; programs; promoter; single-cell RNA sequencing; transcription factor; transcriptome

PROJECT SUMMARY/ABSTRACT During the past decades, studies of heart development have been mainly focused on conserved gene regulatory mechanisms, which control various aspects of cardiogenesis across multiple species from drosophila to mouse. These conserved gene programs include cardiac transcriptional factors (such as Nkx2.5, Isl1, Tbx20), microRNAs, and epigenetic regulators. Despite the high evolutionary conservation of cardiogenesis, the human heart exhibits unique properties, including distinctive morphogenesis and electrophysiological properties. These species-specific differences suggest the existence of novel genetic programs in each species. As the limitation of experimental setting, it remains unclear the mechanisms that regulate unique aspects of human cardiogenesis, and how human-specific mechanisms interact with conserved gene programs to fine-tune human heart development. Recently, accumulating evidence demonstrated that long noncoding RNAs (lncRNAs) play important roles in cell fate specification and organogenesis, including the heart. LncRNAs are greater than 200bp non-coding transcripts, which account for >40% of human transcriptome. Many lncRNAs are tissue-specific and species-specific. Recently, we identified a novel human-specific lncRNA, named Heart Brake LncRNA1 (HBL1) (Dev. Cell. 2017, 4:333-8). HBL1 is highly expressed in both cytoplasm and nucleus of human induced pluripotent stem cells (hiPSCs). Cytosolic HBL1 modulates cardiomyocyte (CM) development from hiPSCs by counteracting microRNA1 (MIR1). Pluripotency marker gene SOX2 activates HBL1 transcription. In this proposal, we will analyze further, a novel mechanism of nuclear HBL1 in initiating the cardiac gene-expressing program via interacting with polycomb repressive complex 2 (PRC2). Additionally, we will test a hypothesis that HBL1 adds a new layer of human- specific regulatory mechanism on top of a conserved cardiogenic axis. All together, the central hypothesis is that transcriptional factor SOX2, lncRNA HBL1, microRNA-1 and PRC2 complex composite a whole network to control human cardiogenesis from pluripotent stem cells.

项目摘要/摘要 在过去的几十年中，心脏发育的研究主要集中在保守的基因上 调节机制，控制着多种物种的心脏病的各个方面 果蝇到小鼠。这些保守的基因程序包括心脏转录因子（例如NKX2.5， ISL1，TBX20），microRNA和表观遗传调节剂。尽管有很高的进化保护 心脏病发生，人类心脏表现出独特的特性，包括独特的形态发生和 电生理特性。这些特定物种的差异表明存在新遗传 每个物种的程序。作为实验环境的局限性，尚不清楚 调节人类心脏病的独特方面，以及人类特异性机制如何与 保守的基因程序，以微调人类心脏的发展。最近，积累了证据 证明长的非编码RNA（LNCRNA）在细胞命运规范和 器官发生，包括心脏。 LNCRNA大于200bp的非编码成绩单，该笔录的解释为 > 40％的人类转录组。许多LNCRNA是组织特异性和物种特异性的。最近，我们确定了 一种新型的人类特异性LNCRNA，名为Heart Brake LncRNA1（HBL1）（Dev。Cell。2017，4：333-8）。 HBL1是 在人类诱导的多能干细胞（HIPSC）的细胞质和核中高度表达。胞质 HBL1通过抵消MicroRNA1（miR1）调节HIPSC的心肌细胞（CM）发育。 多能标记基因SOX2激活HBL1转录。在此提案中，我们将进一步分析一本小说 核HBL1的机理在启动心脏基因表达程序中通过与PolyComb相互作用 抑制性复合物2（PRC2）。此外，我们将检验一个假设，即HBL1添加了一层新的人类 保守的心源轴之上的特定调节机制。总之，中心假设是 转录因子SOX2，LNCRNA HBL1，microRNA-1和PRC2复合复合材料整个网络 从多能干细胞中控制人的心脏病。