Midface and Upper Airway in Craniosynostosis

描述(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顶缝合过早闭合，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良，这是一种包罗万象的诊断，其基础是上颌、鼻咽和颧骨发育不足，表现为面部比平均水平更平，并伴随呼吸困难。我们有来自人类和小鼠的颅缝融合模型的新数据，这些数据详细说明了颅顶、颅底、面部骨骼、面部缝合和上呼吸道的精确方面，这些方面受到各种FGFR2和FGFR3突变的不同影响。我们假设，“面中部发育不全”的全身性症状代表了一种复杂的表型，其组成部分因具体突变而异。对于颅缝融合中被确认为致病基因的突变如何影响发育中的颅面复合体的细胞和组织，从而导致面中部和后鼻孔发育不良或闭锁，人们知之甚少。在这个项目中，我们将对来自 人类颅缝早闭表型，以制定和测试与各种突变对小鼠颅缝早闭模型的局部影响有关的假设。我们将利用条件突变小鼠，包括FGFR2+/S252W和FGFR2+/P253R Apert综合征，Fgfr2cC342Y/+Crouzon/Pfeiffer综合征，FGFR3+/P244R Muenke综合征，以及Wnt1-、Mesp1-和Sox17-2A-I-Cre-Report小鼠，定量研究这些突变对特定头面部结构和细胞和组织的差异影响，这些突变来自不同的胚胎组织，并有助于‘面中部发育不良’。一旦在面部和颅底表型上明确了这些精确的表型效应，我们将分析这些突变在细胞迁移、分化、凋亡和增殖中的作用。最后，我们将利用RNA-SEQ、原位杂交、免疫印迹和免疫组织化学等方法对特定区域和发育阶段的组织进行微观解剖，仔细研究候选的FGFR途径(如ERK1/2、p38)，并确定新的途径。我们将提供有关上呼吸道和面部畸形发生的基本信息，并为面中部和后鼻孔发育不良的定义建立客观标准，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.