Characterizing the mechanisms of PDGFRalpha regulation in upper lip development

表征PDGFRα在上唇发育中的调节机制

• 批准号: 8522826
• 负责人: Fenglei He
• 金额: $ 4.81万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2014-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8522826
• 关键词: Ablation; Affect; Bilateral; Biological Assay; Birth; Cell Culture Techniques; Cell Proliferation; Cell Survival; Cells; Cephalic; Chemicals; Cleaved cell; Cleft Lip; Congenital Abnormality; Data; Defect; Development; Developmental Process; Disease; Dorsal; Eating; Ectoderm; Embryo; Embryonic Development; Exhibits; Face; Future; Genes; Genetic; Goals; High Prevalence; Human; Immunohistochemistry; Impairment; In Vitro; Individual; Investigation; Knock-out; Knockout Mice; Laboratories; Ligands; Link; Lip structure; Maxilla; Medial; Mediating; Mesenchymal; Methods; Migration Assay; Molecular; Morphogenesis; Mus; Mutant Strains Mice; Mutation; Neural Crest; Neural Crest Cell; Newborn Infant; Nose; PDGFA gene; PDGFRB gene; Pathogenesis; Plants; Platelet-Derived Growth Factor; Platelet-Derived Growth Factor Receptor; Population; Prevention; Primordium; Process; Regulation; Research; Role; Signal Pathway; Signal Transduction; Tissues; Transgenic Mice; Transgenic Organisms; Work; cell motility; cleft lip and palate; craniofacial; design; in vitro Assay; in vivo; inhibitor/antagonist; lip morphogenesis; migration; mutant; novel; orofacial; public health relevance; relating to nervous system; research study; spatiotemporal; tool

DESCRIPTION (provided by applicant): Cleft lip is one of most common birth defects. Caused by disruption of normal craniofacial development, cleft lip affects approximately 1 in 500 newborns worldwide. The long term goal of this proposed research is to understand the mechanisms of upper lip development and of orofacial cleft pathogenesis. Mutations of Platelet Derived Growth Factor Receptor ¿ (PDGFR¿) signaling have been tightly linked to cleft lip/palate in humans and mice, suggesting an evolutionarily conserved role in craniofacial development. During embryo development, the upper lip is formed by fusion of medial nasal process (MNP) and maxillary processes, both originating from neural crest cells. This proposed research is aimed at characterizing the role of PDGFR¿ in development of MNP and of neural crest cells during upper lip morphogenesis. First, we will generate a novel transgenic line Alx3Cre to examine the role of PDGFR¿ specifically in MNP development. Cell proliferation and cell survival will be assayed in the MNP of conditional knockout and control embryos using immunohistochemistry methods. Scratch assay and transwell assay will be carried out to analyze the role of PDGFR¿ signaling in MNP cell migration. In the second specific aim, we will examine the role of PDGFR¿ specifically in neural crest cells. Lineage tracing method and in vitro explants culture approaches will be combined with immunohistochemistry experiments in the conditional knockout and control embryos. In the third specific aim, we will analyze the role of PDGFR¿ engaged PI3K signaling in MNP and neural crest development using PDGFR¿ signaling mutant PDGFR¿ PI3K/PI3K mice. In vitro cell migration assays and in vivo mouse genetic studies proposed in first and second specific aims will be carried out on PDGFR¿ PI3K/PI3K and control embryos. Results of the proposed works will reveal the fundamental role of PDGFR¿ in MNP and neural crest cell development during upper lip morphogenesis. The results of proposed research will provide novel information to understand the fundamental mechanisms of MNP and upper lip formation. This study will benefit treatment and prevention of cleft lip in the future, to ultimately reduce the occurrence of this birth defect in newborns.

描述（由申请人提供）：唇裂是最常见的出生缺陷之一。由于正常颅面发育受到破坏，唇裂影响着全世界大约五百分之一的新生儿。这项研究的长期目标是了解上唇发育和口面裂发病机制。血小板衍生生长因子受体 (PDGFR) 信号突变与人类和小鼠的唇裂/腭裂密切相关，表明在颅面发育中具有进化保守的作用。在胚胎发育过程中，上唇是由内侧鼻突（MNP）和上颌突融合形成的，两者都起源于神经嵴细胞。这项研究旨在表征 PDGFR 在上唇形态发生过程中 MNP 和神经嵴细胞发育中的作用。首先，我们将生成一种新型转基因系 Alx3Cre，以检查 PDGFR 在 MNP 发育中的具体作用。将使用免疫组织化学方法在条件敲除胚胎和对照胚胎的 MNP 中测定细胞增殖和细胞存活。将进行划痕实验和transwell实验来分析PDGFR¿信号在MNP细胞迁移中的作用。在第二个具体目标中，我们将特别研究 PDGFR 在神经嵴细胞中的作用。谱系追踪方法和体外外植体培养方法将与条件敲除和对照胚胎中的免疫组织化学实验相结合。在第三个具体目标中，我们将使用 PDGFR¿ 信号突变体 PDGFR¿ PI3K/PI3K 小鼠分析 PDGFR¿ 参与 PI3K 信号传导在 MNP 和神经嵴发育中的作用。第一个和第二个具体目标中提出的体外细胞迁移测定和体内小鼠遗传学研究将在 PDGFR¿PI3K/PI3K 和对照胚胎上进行。拟议工作的结果将揭示 PDGFR¿ 在上唇形态发生过程中 MNP 和神经嵴细胞发育中的基本作用。所提出的研究结果将为理解 MNP 和上唇形成的基本机制提供新的信息。这项研究将有利于未来唇裂的治疗和预防，最终减少新生儿这种出生缺陷的发生。