Growth Factor Signaling and Craniofacial Development

生长因子信号传导和颅面发育

• 批准号: 9981416
• 负责人: Philippe M Soriano
• 金额: $ 59.86万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9981416
• 关键词: Affect; Area; Binding Sites; Biology; Biosensor; Cell Differentiation process; Cell Proliferation; Cells; Cephalic; Cleft Palate; Cleft lip with or without cleft palate; Complex; Congenital Abnormality; Craniofacial Abnormalities; Development; Embryo; Face; Fibroblast Growth Factor; Genes; Genetic Transcription; Goals; Growth Factor; Human; Immediate-Early Genes; Individual; Kinetics; Link; Location; MEKs; Mesenchyme; Molecular; Morphogenesis; Mus; Mutagenesis; Mutant Strains Mice; Mutation; Neural Crest; Neural Crest Cell; Osteoblasts; Output; PC12 Cells; Pathway interactions; Phenotype; Platelet-Derived Growth Factor; Population; Prevention; Process; Receptor Protein-Tyrosine Kinases; Role; Serum Response Factor; Signal Pathway; Signal Transduction; Site; Work; cleft lip and palate; combinatorial; craniofacial; craniofacial development; developmental disease; experimental study; gene induction; genetic analysis; in vivo; inhibitor/antagonist; mutant; myocardin; new technology; novel; programs; receptor; response; transcription factor

The major aims of this proposal are to identify signaling mechanisms underlying midface development that are controlled by PDGF and FGF. Loss of the Pdgfra or Fgfr1 receptors leads to facial clefting and improper development of the frontonasal process, whereas hypomorphic mutations in these pathways result in cleft palate. Pdgfra and Fgfr1 regulate craniofacial development mainly in cranial neural crest cells (cNCCs) through PI3K and Erk, respectively. PDGF induces a short duration or Erk signaling cell differentiation, whereas FGF promotes cell proliferation and perdurance of the Erk signal. Last, PDGF and FGF regulate craniofacial development by engaging immediate early genes (IEGs) through serum response factor (Srf), a critical transcription factor activated by these growth factors itself required for midface closure. This application proposes: 1. To establish the roles and locations of PI3K and Erk signaling in craniofacial development. We will analyze midface development in conditional PI3K and Erk core component genes in NCCs. To identify sites of PDGF and FGF driven signaling activity in vivo, we will breed PI3K and Erk biosensors into wild type, Pdgfr and Fgfr mutant backgrounds. 2. To determine how PDGF and FGF signaling differences differentially regulate craniofacial development. We will alter the duration of Erk signaling in primary MEPMs using Mek/Erk and PKC inhibitors, and investigate how this affects cell differentiation and proliferation. To establish the relative importance of combinatorial vs. dynamic signaling and downstream responses in craniofacial development, we will analyze expression of known PDGF and FGF transcriptional targets that are PI3K and Erk dependent and linked to differentiation or proliferative responses, in Pdgfr/PI3K or Fgfr/Erk neural crest specific mutants. 3. To determine the signaling mechanisms through which Srf, a shared PDGF and FGF transcriptional target, regulates differential transcriptional outputs. Srf interacts with two classes of co-factors, Myocardin Related Transcription Factors (MRTFs) or Ternary Complex Factors (TCFs). PDGF promotes the association of Srf with MRTFs through PI3K to regulate the expression of cytoskeletal target genes critical for craniofacial development, whereas both PDGF and FGF allow Srf to interact with TCFs through PI3K and Erk signaling to facilitate the expression of core IEGs. To establish the molecular pathways and targets by which growth factors regulate midface development through Srf, we will generate mice carrying mutations in Srf that abrogate its ability to associate with MRTFs, while maintaining its interactions with TCFs. These proposed studies explore novel territories in the area of growth factor signaling in craniofacial biology and open new directions for the prevention of craniofacial birth defects.

该提案的主要目的是确定中间开发基础的信号传导机制是 由PDGF和FGF控制。 PDGFRA或FGFR1受体的丧失导致面部裂纹和不适当 额骨过程的发展，而这些途径中的型突变导致口感cle裂。 PDGFRA和FGFR1主要通过PI3K调节颅面开发（CNCCS） 和ERK。 PDGF诱导持续时间短或ERK信号传导细胞分化，而FGF促进 ERK信号的细胞增殖和透明度。最后，PDGF和FGF通过 通过血清反应因子（SRF）参与即时的早期基因（IEG），这是关键转录因子 这些生长因子本身被激活的中心封闭所需的。本申请提出： 1。建立PI3K和ERK信号传导在颅面发育中的作用和位置。我们将分析 NCC中有条件PI3K和ERK核心成分基因中的中间发育。识别PDGF的位置 在体内，FGF驱动的信号传导活性，我们将在野生型，PDGFR和FGFR中繁殖PI3K和ERK生物传感器 突变背景。 2。确定PDGF和FGF信号传导差异如何差异地调节颅面发育。我们 将使用MEK/ERK和PKC抑制剂在初级MEPM中改变ERK信号的持续时间，并研究如何 这会影响细胞分化和增殖。建立组合与动态的相对重要性 颅面发育中的信号传导和下游反应，我们将分析已知PDGF的表达 以及依赖于PI3K和ERK的FGF转录靶标，与分化或增殖有关 响应，在PDGFR/PI3K或FGFR/ERK神经rest特定突变体中。 3。确定SRF，共享PDGF和FGF转录目标的信号传导机制， 调节差分转录输出。 SRF与两个类别的辅助因素相互作用，与心肌蛋白相关 转录因子（MRTF）或三元复合因子（TCF）。 PDGF促进了SRF与 MRTF通过PI3K调节细胞骨架靶基因的表达对于颅面发育至关重要， 而PDGF和FGF均允许SRF通过PI3K和ERK信号与TCF相互作用，以促进 核心IEG的表达。建立分子途径和靶标的生长因子调节的靶标 通过SRF开发中间，我们将生成携带SRF突变的小鼠，以消除其能力 与MRTF相关联，同时保持其与TCF的相互作用。 这些拟议的研究探讨了颅面生物学和 开放预防颅面先天缺陷的新方向。