Growth Factor Signaling and Craniofacial Development

生长因子信号传导和颅面发育

• 批准号: 10226072
• 负责人: Philippe M Soriano
• 金额: $ 60.73万
• 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/10226072
• 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.

这项建议的主要目的是确定面中部发育的信号机制， 由PDGF和FGF控制。Pdgfra或Fgfr 1受体的缺失会导致面部皲裂和不当的 在额鼻突的发育过程中，这些通路中的亚形态突变导致腭裂。 Pdgfra和Fgfr 1主要通过PI 3 K在颅神经嵴细胞（cNCCs）中调节颅面发育 和Erk，分别。PDGF诱导短时间或Erk信号传导细胞分化，而FGF促进细胞分化。 细胞增殖和Erk信号的持久性。最后，PDGF和FGF调节颅面发育， 通过血清反应因子（Srf），一种关键的转录因子， 由这些生长因子本身激活，这是面中部闭合所必需的。本申请提出： 1.探讨PI 3 K和Erk信号通路在颅颌面发育中的作用和定位。我们将分析 NCC中条件性PI 3 K和Erk核心组分基因的面中部发育。确定PDGF的位点 和FGF驱动的体内信号传导活性，我们将PI 3 K和Erk生物传感器培育成野生型、Pdgfr和Fgfr 变种人背景 2.确定PDGF和FGF信号差异如何调节颅面发育。我们 将使用Mek/Erk和PKC抑制剂改变原发性MEPMs中Erk信号传导的持续时间，并研究如何 这影响细胞分化和增殖。确定组合与动态的相对重要性 在颅面发育的信号和下游反应，我们将分析已知的PDGF的表达， 和FGF转录靶，其是PI 3 K和Erk依赖性的并且与分化或增殖相关， 反应，在Pdgfr/PI 3 K或Fgfr/Erk神经嵴特异性突变体。 3.为了确定Srf，一个共同的PDGF和FGF转录靶点， 调节差异转录输出。SRF与两类辅因子相互作用， 转录因子（MRTF）或三元复合物因子（TCF）。PDGF促进Srf与 MRTFs通过PI 3 K调节颅面发育关键细胞骨架靶基因的表达， 而PDGF和FGF都允许Srf通过PI 3 K和Erk信号传导与TCF相互作用，以促进TCF的增殖。 核心IEGs的表达。建立生长因子调控的分子途径和靶点 通过Srf的中面发育，我们将产生携带Srf突变的小鼠，这些突变消除了其 与MRTF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中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的方向。