Function and Regulation of Sema3 Genes in Palate Development and Innervation

Sema3 基因在上颚发育和神经支配中的功能和调节

基本信息

批准号：
10380003
负责人：
Yu Lan
金额：
$ 19.88万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10380003
关键词：
Address Affect Anatomy Bilateral Binding Proteins Bioinformatics Biology CRISPR/Cas technology Caring ChIP-seq Child Chromosomal Rearrangement Cleft Palate Complex Congenital Abnormality Counseling Cranial Nerves Data Developed Countries Development Developmental Process Diagnosis Disease Dysarthria Embryo Endothelium Exhibits Face Facial paralysis Family Functional disorder Gene Family Genes Genetic study Genomic Segment Growth Human Impairment Knowledge Lead Link Live Birth Maps Mediating Mediator of activation protein Mesenchyme Missense Mutation Molecular Morphogenesis Mus Mutant Strains Mice Mutation Neural Crest Neural Crest Cell Neuraxis Neurologic Dysfunctions Neurons Nose Online Mendelian Inheritance In Man Operative Surgical Procedures Palate Pathogenesis Pathogenicity Pathway interactions Patient Care Patients Penetrance Play Proteins Quality of life Regulation Regulatory Element Reporting Research Research Project Grants Role SHH gene Semaphorin-3A Series Signal Transduction Speech Structural Congenital Anomalies Syndrome Testing Tissues Transcriptional Regulation Zinc Fingers axon growth axon guidance cell motility craniofacial experience feeding genome editing improved malformation member migration mortality mouse genetics mouse model multiple omics mutant nerve supply novel orofacial osteoblast differentiation osteogenic palatal shelves palate repair palatogenesis receptor repaired transcription factor transcriptome transcriptome sequencing vasculogenesis

项目摘要

Abstract Cleft palate is a common birth defect in humans and causes significant problems in feeding, speech, and increased mortality. While most cleft palate patients receive surgical repair to restore an anatomically intact palate, about 30% of children with a repaired palate still experience significant functional palate deficit and/or velopharyngeal dysfunction (VPD), which impairs speech and significantly affects quality of life. Orofacial malformations, including cleft or high-arched palate and flattened nasal bridge, and nasal dysarthria are major features of congenital facial palsy conditions such as the Moebius syndrome (OMIM 157900), suggesting that facial/palatal innervation is intimately linked to palate morphogenesis and velopharyngeal function. However, nothing is currently known about the molecular mechanisms regulating palate innervation and their integration with the overall mechanisms regulating palate morphogenesis. We have discovered that mice lacking the zinc finger transcription factor Osr2 exhibit complete cleft palate and dramatic disruption of palatal innervation. We previously demonstrated that Osr2 is a key intrinsic regulator of palatal shelf growth and integrates the functions of several major molecular pathways, including SHH signaling and the transcription factor Pax9- mediated regulation of palate morphogenesis. Through RNA-seq transcriptome profiling we found that expression of three physically linked members of the Sema3 gene family, Sema3a, Sema3d, and Sema3e, was significantly upregulated in the developing palatal mesenchyme in Osr2 mutant embryos. The Sema3 proteins were originally identified as negative mediators of axonal guidance in the central nervous system and have since been shown to play crucial roles in many other developmental processes, including neural crest migration, endothelial migration and vasculogenesis, and osteoblast differentiation. Our central hypothesis is that proper regulation of expression of the Sema3 genes during palate development is crucial for not only palate innervation but also palate morphogenesis. Recently mutations in PLXND1, which encodes a receptor for SEMA3 proteins, and a complex chromosomal rearrangement disrupting both the SEMA3A and SEMA3D genes, have been associated with Moebius syndrome, suggesting that the SEMA3-PLXND1 pathway plays crucial roles in cranial nerve development and palate morphogenesis in humans as well. To better understand the function and regulation of the Sema3 genes in palate development and innervation, we will use a combination of multi-omics approaches, CRISPR/Cas9-mediated genome editing, and compound mutant mouse studies to unravel the molecular network involving Osr2 and the Sema3 genes in the regulation of palate innervation and morphogenesis. Data from these studies will fill a long standing major gap in the understanding of pathogenic mechanisms of cleft palate and VPD, which will help to significantly improve strategies for diagnosis, counseling, treatment and care of patients with these devastating craniofacial conditions.

抽象的 left裂是人类常见的先天缺陷，在喂养，言语和死亡率增加。虽然大多数口味患者接受手术修复以恢复解剖学完整口感，大约30％的修复味觉的儿童仍然经历着明显的功能性味觉赤字和/或尾咽功能障碍（VPD），会损害语音并显着影响生活质量。口面畸形，包括裂口或高竖立的pa和扁平的鼻桥，以及鼻构障先天性面部麻痹状况（例如Moebius综合征（OMIM 157900））的特征，表明面部/pal骨神经与唇形形态发生和尾咽功能密切相关。然而，目前对调节口感神经支配及其整合的分子机制尚无带有调节味觉形态发生的总体机制。我们发现缺乏锌的老鼠手指转录因子OSR2表现出完全的left裂和pa裂神经的戏剧性破坏。我们以前证明OSR2是palatal架子增长的关键内在调节剂，并将其整合几种主要分子途径的功能，包括SHH信号传导和转录因子PAX9- 介导的味觉形态发生调节。通过RNA-Seq转录组分析，我们发现 Sema3基因家族的三个物理联系成员的表达，Sema3a，Sema3d和Sema3e 在OSR2突变体胚胎中发育中的palatal间充质中显着上调。 SEMA3 蛋白质最初被识别为中枢神经系统中轴突引导的负介体此后已被证明在包括神经在内的许多其他发展过程中扮演着重要角色波峰迁移，内皮迁移和血管生成以及成骨细胞分化。我们的中心假设是，在pa发发育过程中，适当调节Sema3基因的表达至关重要不仅对神经神经，而且还可以触诊形态发生。 plxnd1的最近突变，该突变编码 SEMA3蛋白的受体，以及一个复杂的染色体重排，破坏了SEMA3A和 SEMA3D基因与Moebius综合征有关，表明SEMA3-PLXND1 途径在人类的颅神经发育和pa酸的形态发生中起着至关重要的作用。到更好地了解Sema3基因在pa发育和神经上的功能和调节，我们将结合多种词的方法，CRISPR/CAS9介导的基因组编辑和化合物突变小鼠研究以揭示涉及OSR2和SEMA3基因的分子网络口感神经和形态发生。这些研究的数据将填补长时间的主要差距了解left裂和VPD的致病机制，这将有助于显着改善这些毁灭性颅面患者的诊断，咨询，治疗和护理策略状况。