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 患有的先天性疾病，包括Di George/22Q11DS，Noonan及相关综合症， 心脏缺陷和颅面畸形。 22q11删除综合征来自同名 导致TBX1单倍弥补的删除，而Noonan和相关综合症是影响的。 RAS-MAPK信号通路。但是，了解联合心形的病因 缺陷需要深入了解基因功能的细胞和发育环境。在早期的羊膜中 胚胎，第二个心脏场（SHF）和分支/咽头肌肉从一个共同的 心咽中胚层中多核祖细胞的种群。 tbx1是思想功能 心咽祖细胞，以控制咽部肌发生和SHF的发育，并与 成纤维细胞生长因子（FGF）-MAPK信号在心脏发育过程中，强调了重要性 在早期心咽发育的细胞环境中研究TBX1和FGF-MAPK信号传导。 剪裁ciona成为一种简单而强大的核核模型，用于研究早期心脏咽喉 开发，具有高空间和时间分辨率。在Ciona，四个多能心咽 祖细胞经历刻板印象的迁移和细胞分裂，产生独特的第一和第二心脏，以及 咽部肌肉谱系部署了用脊椎动物保守的基因网络。利用唯一的 Ciona系统的优势，以及使用谱系特异性扰动的大量先前工作，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模式的建立。拟议的工作将解决如何 动态心咽中的利基市场有助于极化MAPK信号传导和TBX1/10激活；如何固有 有丝分裂主轴定位的决定因素，Rhogap depdc1有助于祖细胞向祖细胞定向分裂 关于利基市场，并分析MAPK信号和MAPK信号之间的拮抗作用的分子基础 早期心脏计划。其次，该提案将探索之间的时间动态 心咽状状态，通过研究新的基因表达和命运选择与细胞的结合 分裂以及成熟祖细胞中的转录组变化如何决定 心脏咽中的祖细胞形成心脏和咽肌肉前体。完成这个雄心勃勃的 提案将产生对心血管广泛意义的新兴概念的深远见解 发育和干细胞生物学。