Cellular determinants of cardiopharyngeal multipotency and early fate choices

心咽多能性和早期命运选择的细胞决定因素

• 批准号: 9981188
• 负责人: Lionel Christiaen
• 金额: $ 61.3万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9981188
• 关键词: Address; Affect; Bar Codes; CRISPR/Cas technology; Cardiac; Cardiovascular system; Cell Count; Cell division; Cells; Chordata; Competence; Congenital Heart Defects; Coupled; Coupling; Craniofacial Abnormalities; DNA; Development; DiGeorge Syndrome; Disease; Down-Regulation; Embryo; Ephrins; Etiology; Exhibits; Fibroblast Growth Factor; GATA4 gene; Gene Activation; Gene Expression; Genes; Genetic Transcription; Genomic approach; Genomics; Head; Heart; Heart Abnormalities; Lateral; MAP Kinase Gene; MAPK Signaling Pathway Pathway; Maps; Mediating; Mesoderm; Methods; Mitotic spindle; Modeling; Molecular; Multipotent Stem Cells; Muscle; Mutagenesis; Myocardium; Newborn Infant; Noonan Syndrome; Pattern; Phosphoric Monoester Hydrolases; Population; Positioning Attribute; Regulation; Resolution; Role; Signal Transduction; Stereotyping; Syndrome; System; Testing; Up-Regulation; Urochordata; Vertebrates; Work; base; cardiogenesis; cell type; craniofacial; gene function; genome-wide; genomic data; inhibitor/antagonist; innovation; insight; migration; myogenesis; pharynx muscle; progenitor; programs; quantitative imaging; rho GTPase-activating protein; single-cell RNA sequencing; stem cell biology; stem cells; temporal measurement; transcriptome

SUMMARY Prevalent congenital diseases, including the Di George/22q11DS, Noonan and related syndromes, present with both cardiac defects and craniofacial dysmorphism. The 22q11 Deletion Syndrome arises from eponymous deletions that cause TBX1 haploinsufficiency, while Noonan and related syndromes are RASopathies that affect the RAS-MAPK signaling pathway. However, understanding the etiology of combined cardio-craniofacial defects requires insights into the cellular and developmental contexts of gene function. In early amniote embryos, the second heart field (SHF) and branchiomeric/pharyngeal head muscles emerge from a common population of multipotent progenitors in the cardiopharyngeal mesoderm. TBX1 is thought function in cardiopharyngeal progenitors, to control both pharyngeal myogenesis and SHF development, and interact with Fibroblast Growth Factor (FGF)-MAPK signaling during heart development, highlighting the importance of studying Tbx1 and FGF-MAPK signaling in the cellular context of early cardiopharyngeal development. The tunicate Ciona emerged as a simple and powerful chordate model to study early cardiopharyngeal development, with high spatial and temporal resolution. In Ciona, four multipotent cardiopharyngeal progenitors undergo stereotyped migration and cell divisions, producing distinct first and second cardiac, and pharyngeal muscle lineages that deploy gene networks conserved with vertebrates. Leveraging the unique strengths of the Ciona system, and extensive previous work using lineage-specific perturbations, including CRISPR/Cas9-mediated mutagenesis, quantitative imaging, and multiplexed single cell genomics methods, this proposal will first explore the establishment of spatial patterns. The proposed work will address how a dynamic cardiopharyngeal niche helps polarize MAPK signaling and Tbx1/10 activation; how an intrinsic determinant of mitotic spindle positioning, the RhoGAP Depdc1, helps progenitors orient their divisions with regards to the niche, and analyze the molecular basis for the antagonism between MAPK signaling and the early heart program. Second, this proposal will explore the temporal dynamics underlying transitions between cardiopharyngeal states, by studying how de novo gene expression and fate choices are coupled with cell divisions, and how transcriptome changes in maturing progenitors determine the competence of cardiopharyngeal progenitors to form heart and pharyngeal muscle precursors. Completion of this ambitious proposal will yield far-reaching insights into emerging concepts of broad significance for cardiovascular developmental and stem cell biology.

概括 流行的先天性疾病，包括 Di George/22q11DS、Noonan 和相关综合征，表现为 心脏缺陷和颅面畸形。 22q11 缺失综合症源自同名 导致 TBX1 单倍体不足的缺失，而 Noonan 和相关综合征是影响 RASopathies RAS-MAPK 信号通路。然而，了解心颅面联合病变的病因学 缺陷需要深入了解基因功能的细胞和发育背景。在羊膜早期 胚胎、第二心脏区（SHF）和鳃头/咽头肌肉来自共同的 心咽中胚层中的多能祖细胞群。 TBX1 是思想函数 心咽祖细胞，控制咽肌发生和 SHF 发育，并与 成纤维细胞生长因子 (FGF)-MAPK 信号在心脏发育过程中的重要性 研究早期心咽发育细胞环境中的 Tbx1 和 FGF-MAPK 信号传导。 被囊动物 Ciona 作为一种简单而强大的脊索动物模型出现，用于研究早期心咽 发展，具有高空间和时间分辨率。在 Ciona 中，四个多能心咽 祖细胞经历定型的迁移和细胞分裂，产生不同的第一和第二心脏，并且 部署脊椎动物保守的基因网络的咽肌谱系。凭借独特的 Ciona 系统的优势，以及之前使用谱系特定扰动进行的大量工作，包括 CRISPR/Cas9介导的诱变、定量成像和多重单细胞基因组学方法， 该提案将首先探讨空间格局的建立。拟议的工作将解决如何 动态心咽生态位有助于极化 MAPK 信号传导和 Tbx1/10 激活；如何内在的 有丝分裂纺锤体定位的决定因素 RhoGAP Depdc1 帮助祖细胞定向其分裂 针对生态位，分析MAPK信号通路与MAPK信号通路拮抗的分子基础 早期心脏计划。其次，该提案将探讨之间的转换背后的时间动态 心咽状态，通过研究从头基因表达和命运选择如何与细胞耦合 分裂，以及成熟祖细胞的转录组变化如何决定细胞的能力 心咽祖细胞形成心脏和咽肌前体。完成这个宏伟的目标 该提案将对心血管领域具有广泛意义的新兴概念产生深远的见解 发育和干细胞生物学。