Genetic, Tissue, and Anatomical Interactions in Mandibulofacial Dysmorphogenesis

项目总结 面部畸形总是需要某种类型的治疗、矫正手术和密切随访，同时 给患者及其家人带来经济和情感上的负担。尽管对人类数据的分析 人体研究带来了特别的问题，尤其是产前的关键时刻 发展是不可用来研究的。这一建议旨在确定发育和分子水平上的 利用我们通过系统鉴定的小鼠突变体处理潜在的下颌面部异常 检索国际小鼠表型鉴定联盟(IMPC)的最新内容作为回应 NIH PAR-20-137用于IMPC胚胎和围产期致死的KO小鼠品系的表型鉴定。微型-或 下颌后畸形是描述颌面部发育不良的下颌表型最常用的术语， 然而，目前在诊断下颌骨畸形方面缺乏精确度并没有批判性地考虑到 这些情况的潜在不同病因及其对异常的潜在后遗症的影响。 小颌症描述的是下颌骨绝对缩小，这表明下颌骨主要是 受影响的，而下颌后突指的是正常大小的下颌骨，相对于上颌骨放置在后面 下巴。因此，小下颌和后下颌，虽然提供相似的面部轮廓，但由不同的原发 每个发育过程都可能与舌头和腭部发育有不同的结合。什么时候 下颌畸形会出现舌下垂、呼吸道阻塞，在某些情况下，还会出现腭裂， 这种情况被称为皮埃尔·罗宾(PR)。一系列发育性疾病的致病机制 PR的事件一直被假设，但发现的突变之间几乎没有明确的因果关系， 失调的基因表达，精确的细胞过程，以及PR相关的异常都被记录在案。至 验证这一假设，我们计划了一套仔细协调和充分合作的IMPC突变小鼠的分析 根据已知的导致人类PR的基因或PR特征的存在来鉴定品系 记录在这些鼠标线上。我们的深入表型将涉及：目标1：定量三维形态 用磷钨酸增强的显微计算机断层摄影术对胚胎进行分析.AIM2：鉴别 突变前后相关组织和发育时间点的基因表达分析 利用散装rna-seq和空间转录组学技术控制乳房虫；目的3：使用原位组织学研究 相关基因、信号通路、细胞过程和 分化状态以确定导致细胞畸形发生的细胞和分子事件 下颌面复合体。这种多层次的系统生物学方法将提供对 基因变异对下颌及相关舌、腭部和上呼吸道表型的局部影响 利用建立模型的小鼠品系，确定与它们的生产有关的发育和分子功能 与这些病症相关的表型。

PROJECT SUMMARY Anomalies of the face invariably require some type of therapy, corrective surgery, and close follow-up while imposing a financial and emotional burden on patients and their families. Although the analysis of human data is critical, human studies pose particular problems, not the least of which is that critical times of prenatal development are not available for study. This proposal aims to identify the developmental and molecular processes underlying mandibulofacial anomalies using mouse mutants that we identified by systematically searching the current contents of the International Mouse Phenotyping Consortium (IMPC) in response to NIH PAR-20-137 for phenotyping IMPC embryonic and perinatal lethal KO mouse lines. Micro- or retrognathia are the most common terms used to describe mandibular phenotypes in mandibulofacial dysostosis, yet the current lack of precision in diagnoses of mandibular dysmorphology does not critically consider the potentially distinct etiology of these conditions and their influence on potential sequelae of anomalies. Micrognathia describes a mandible that is absolutely reduced in size, indicating that the mandible is primarily affected, while retrognathia refers to a normally sized mandible that is placed posteriorly relative to the upper jaw. Thus, micrognathia and retrognathia, while providing similar facial profiles, are produced by different primary developmental processes and each may integrate differently with tongue and palatal development. When mandibular dysmorphology occurs with glossoptosis, respiratory obstruction, and in some cases, a cleft palate, the condition is referred to as Pierre Robin (PR). A causative pathogenesis of a sequence of developmental events has been hypothesized for PR, but few clear causal relationships between discovered mutations, dysregulated gene expression, precise cellular processes, and PR-associated anomalies are documented. To test this hypothesis, we plan a carefully coordinated and fully collaborative set of analyses of IMPC mutant mouse lines identified based on genes known to be causative for PR in humans or on the presence of PR features recorded in these mouse lines. Our in-depth phenotyping will involve: Aim 1: quantitative 3D morphologic analyses of embryos using phosphotungstic acid-enhanced micro computed tomography; Aim2: differential gene expression analysis of relevant tissues and developmental time points between mutant and unaffected littermate controls using bulk RNA-seq and spatial transcriptomics; Aim 3: histologic studies using in situ hybridization or immunohistochemistry of relevant genes, signaling pathways, cellular processes, and differentiation states to determine the cellular and molecular events giving rise to dysmorphogenesis of the mandibulofacial complex. This multi-level, systems biology approach will provide precise definitions of the localized effects of genetic variants on mandibular and associated tongue, palatal, and upper airway phenotypes to identify the developmental and molecular functions involved in their production, using mouse lines that model the phenotypes associated with these conditions.