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Growth Factor Signaling and Craniofacial Development

生长因子信号传导和颅面发育

基本信息

批准号：
10461030
负责人：
Philippe M Soriano
金额：
$ 61.01万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10461030
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 mutant myocardin new technology novel programs receptor response transcription factor

项目摘要

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面部发育背后的信号机制由PDGF和FGF控制。 Pdgfra 或 Fgfr1 受体的缺失会导致面部裂痕和不当行为额鼻过程的发育，而这些途径的亚等位性突变会导致腭裂。 Pdgfra和Fgfr1主要通过PI3K在颅神经嵴细胞（cNCC）中调节颅面发育和埃尔克，分别。 PDGF 诱导短期或 Erk 信号细胞分化，而 FGF 则促进细胞增殖和 Erk 信号的持久性。最后，PDGF 和 FGF 通过以下方式调节颅面发育：通过血清反应因子 (Srf)（一种关键的转录因子）参与即早期基因 (IEG) 由中面部闭合所需的这些生长因子本身激活。本申请提出： 1. 确定PI3K和Erk信号在颅面发育中的作用和位置。我们将分析 NCC 中条件 PI3K 和 Erk 核心成分基因的中面部发育。鉴定 PDGF 的位点和 FGF 驱动体内信号活动，我们将把 PI3K 和 Erk 生物传感器培育成野生型、Pdgfr 和 Fgfr 突变体背景。 2. 确定PDGF和FGF信号差异如何差异调节颅面发育。我们将使用 Mek/Erk 和 PKC 抑制剂改变原代 MEPM 中 Erk 信号传导的持续时间，并研究如何这会影响细胞分化和增殖。确定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 PI3K 和 Erk 依赖性且与分化或增殖相关的 FGF 转录靶点 Pdgfr/PI3K 或 Fgfr/Erk 神经嵴特异性突变体中的反应。 3. 确定 Srf（PDGF 和 FGF 共同转录靶标）通过的信号传导机制调节差异转录输出。 Srf 与两类辅助因子相互作用，即心肌素相关因子转录因子 (MRTF) 或三元复合因子 (TCF)。 PDGF 促进 Srf 与 MRTF通过PI3K调节对颅面发育至关重要的细胞骨架靶基因的表达，而 PDGF 和 FGF 都允许 Srf 通过 PI3K 和 Erk 信号与 TCF 相互作用，以促进核心 IEG 的表达。建立生长因子调节的分子途径和靶点通过 Srf 进行中面部发育，我们将培育出携带 Srf 突变的小鼠，从而消除了其发育能力与 MRTF 建立联系，同时保持与 TCF 的互动。这些拟议的研究探索了颅面生物学和生长因子信号传导领域的新领域为预防颅面出生缺陷开辟新方向。