Midface and Upper Airway in Craniosynostosis

描述（由申请人提供）：颅缝早闭是一种常见的出生缺陷，可作为综合征的一部分或作为孤立的异常发生。虽然通常被定义为颅穹窿缝的过早闭合，但解剖学和临床研究揭示了整个头部的复杂畸形。颅缝早闭综合征的一个典型的复杂特征是“面中部发育不全”，这是一种基于上颌、鼻咽和颧骨发育不足而定义的综合诊断，表现为面部比平均水平平坦，并伴有呼吸困难。我们有来自人类和小鼠颅缝早闭模型的新数据，这些数据详细描述了受各种FGFR 2和FGFR 3突变不同影响的颅穹窿、颅底、面部骨骼、面部缝和上气道的精确方面。我们推测，广义条件的面中部发育不全，代表了一个复杂的表型，其组成部分不同，取决于特定的突变。相对而言，很少有人知道的是，在颅缝早闭症的病因鉴定的突变如何影响细胞和组织的发展颅面复合体，导致面中部和后鼻孔发育不全或闭锁。在这个项目中，我们将执行3D图像数据的定量分析， 人类颅缝早闭表型，以制定和测试关于各种突变在具有颅缝早闭的小鼠模型中的局部效应的假设。我们将使用条件突变小鼠，包括Fgfr 2 +/S252 W和Fgfr 2 +/P253 R Apert综合征、Fgfr 2cC 342 Y/+ Crouzon/Pfeiffer综合征和Fgfr 3 +/P244 R Muenke综合征，以及Wnt 1-，Mesp 1-和Sox 17 -2A-I-Cre-报告小鼠，以定量研究这些突变对特定颅面结构以及来源于不同胚胎组织的细胞和组织的不同影响，导致“面中部发育不全”一旦这些精确的表型效应在面部和颅底表型上被定义，我们将分析这些突变对细胞迁移、分化、凋亡和增殖的作用。最后，在从特定区域和发育阶段显微解剖的组织上使用RNA-seq、原位杂交、免疫印迹和免疫组织化学，我们将仔细检查候选Fgfr通路（例如，Erk 1/2，p38），并确定新的途径。我们将提供上呼吸道和面部畸形的基本信息，并建立面中部和后鼻孔发育不全的客观标准，以改善许多患有这些疾病的患者的医疗保健。

DESCRIPTION (provided by applicant): Craniosynostosis is a common birth defect that can occur as part of a syndrome or as an isolated anomaly. Though often defined as the premature closure of a cranial vault suture, anatomic and clinical studies reveal complex dysmorphology of the entire head. One complex trait typical of craniosynostosis syndromes is 'midfacial hypoplasia', a catch-all diagnosis that is defined on the basis of deficient development of the upper jaw, nasopharynx, and cheekbones presenting with a flatter than average face and associated respiratory difficulties. We have novel data from humans and mouse models of craniosynostosis which detail precise aspects of the cranial vault, cranial base, facial skeleton, facial sutures, and upper airway that are differentially affected by various FGFR2 and FGFR3 mutations. We hypothesize that the generalized condition of 'midfacial hypoplasia' represents a complex phenotype whose component parts differ depending upon the specific mutation. Relatively little is known about how the mutations identified as causative in craniosynostosis affect cells and tissues of the developing craniofacial complex to result in midfacial and choanal hypoplasia or atresia. In this project, we will perform quantitative analysis of 3D image data from human craniosynostosis phenotypes to formulate and test hypotheses pertaining to localized effects of various mutations in mouse models with craniosynostosis. We will use conditional mutant mice including Fgfr2+/S252W and Fgfr2+/P253R Apert syndrome, Fgfr2cC342Y/+ Crouzon/Pfeiffer syndrome, and Fgfr3+/P244R Muenke syndrome, and Wnt1-, Mesp1- and Sox17-2A-I-Cre-reporter mice to quantitatively study the differential effects of these mutations on specific craniofacial structures and cells and tissues that are derived from diverse embryonic tissues and that contribute to 'midfacial hypoplasia'. Once these precise phenotypic effects are defined on facial and cranial base phenotypes, we will analyze the role of these mutations on cell migration, differentiation, apoptosis, and proliferation. Finally, using RNA-seq, in situ hybridization, immunoblots, and immunohistochemistry on tissues micro-dissected from specific regions and developmental stages, we will scrutinize candidate Fgfr pathways (e.g., Erk1/2, p38) and identify novel pathways. We will provide essential information on dysmorphogenesis of the upper airway and face and establish objective criteria for the definition of midfacial and choanal hypoplasia needed to improve healthcare for numerous patients with these conditions.