Transcriptome Atlases of the Craniofacial Sutures

颅面缝合线转录组图谱

基本信息

批准号：
8847320
负责人：
Greg Peter Holmes
金额：
$ 57.98万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-05-01 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8847320
关键词：
Accounting Adult Affect Apert syndrome Appearance Atlases Biology Bone Development Brain Breathing Breeding Calvaria Candidate Disease Gene Cell Lineage Cells Cephalic Chotzen Syndrome Clinical Management Congenital Abnormality Craniofacial Abnormalities Craniosynostosis Data Data Set Databases Defect Development Disease Elements Embryo Embryology Etiology Evolution Exhibits Eye FGFR2 gene Face Foundations Future Gene Expression Gene Expression Profile Genes Genetic Genetic Predisposition to Disease Goals Growth Head Health Hearing Human Joint structure of suture of skull Joints Knowledge Lead Live Birth Maintenance Mastication Maxilla Mesenchymal Mesenchyme Modeling Molecular Molecular Profiling Morphology Mus Mutant Strains Mice Mutate Mutation Neurologic Nose Operative Surgical Procedures Pathogenesis Pathology Pathway interactions Physical condensation Population Predisposition Process Proliferating RNA Research Resources Skeletal Development Skeleton Staging Surgical Management Surgical sutures Technology Therapeutic Time Tissues Translating Variant Vertebrate Biology adverse outcome base bone cell type coronal suture craniofacial craniofacial development cranium feeding gene discovery insight intramembranous bone intramembranous bone formation laser capture microdissection mouse model mutant mouse model next generation sequencing null mutation osteogenic osteoprogenitor cell postnatal premature prenatal prevent respiratory skull base suture fusion transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): Craniofacial sutures are the fibrous joints between bones, allowing growth of the skull from prenatal to postnatal development until adult size is achieved. Proper suture development is crucial because abnormal suture fusion can require major surgical intervention to restore a satisfactory head and facial appearance and to prevent secondary damage to the brain, eyes, hearing, breathing, and mastication. Craniosynostosis, the premature fusion of skull sutures, is a common birth defect, occurring in 1/2500 live births. I may present in syndromic and non-syndromic forms, and while mutations in some of the genes that account for syndromic forms are known, the underlying genetic etiology has not been identified for the majority of cases that are non- syndromic and involve a single suture. A more comprehensive understanding of suture biology and pathology can be aided by knowledge of gene expression profiles of sutures. Craniofacial sutures vary widely in form, function, and susceptibility to fusion, suggesting that gene expression profiles vary considerably among sutures and during different developmental stages. A detailed characterization of gene expression would require the extraction of specific populations of cells from the different subregions of each suture, including the non-ossifying suture mesenchyme and the flanking osteogenic bone fronts, which are often from distinct bones and may therefore have distinct gene expression patterns. Our overall goal is to generate comprehensive gene expression atlases of the major and functionally important craniofacial sutures of the mouse, which will accelerate both our understanding of human suture biology and the discovery of candidate genes whose mutation may cause craniosynostosis or other defects of craniofacial bone development. We will apply the state-of-the-art technology of laser capture microdissection to obtain tissue from different craniofacial sutures of both normal and craniosynostotic mouse models, combined with next generation sequencing of extracted RNA (RNA-Seq). In Aim 1 we will breed a mouse model of Apert syndrome craniosynostosis with the Fgfr2 S252W mutation and use laser capture microdissection to obtain cells from 11 craniofacial sutures from WT and mutant mice. A second mouse model for Saethre-Chotzen syndrome with a Twist1 heterozygous null mutation will be bred to provide a comparison for two major sutures. In Aim 2 we will extract RNA from the different sutural subregions of WT mice and perform RNA-Seq to generate a comprehensive set of gene expression atlases for normal sutures. In Aim 3 we will similarly extract RNA from the suture subregions of Apert and Saethre-Chotzen syndrome mice for RNA-Seq and generate gene expression atlases complementary to the normal gene expression atlases. These atlases will allow the rapid discovery of genes not yet known to be expressed in sutures, reveal the commonalities and differences between sutures that may suggest new hypotheses of suture formation and differentiation, with wider significance for evolutionary studies of the vertebrate skull, and provide insight into the pathology of suture fusion.

描述（由申请人提供）：颅面缝合线是骨骼之间的纤维关节，允许颅骨从产前到产后的生长直至达到成人尺寸。正确的缝合线发育至关重要，因为异常的缝合线融合可能需要进行重大手术干预才能恢复令人满意的头部和面部外观，并防止对大脑、眼睛、听力、呼吸和咀嚼造成二次损伤。颅缝早闭，即颅骨缝线过早融合，是一种常见的出生缺陷，发生于 1/2500 的活产儿中。 I 可能以综合征和非综合征形式出现，虽然已知一些导致综合征形式的基因突变，但对于大多数非综合征且涉及单缝线的病例，尚未确定潜在的遗传病因学。了解缝线基因表达谱的知识可以帮助更全面地了解缝线生物学和病理学。颅面缝线在形式、功能和融合敏感性方面差异很大，这表明不同缝线和不同发育阶段的基因表达谱差异很大。基因表达的详细表征需要从每个缝线的不同亚区域提取特定的细胞群，包括非骨化缝线间充质和侧翼成骨骨前部，它们通常来自不同的骨骼，因此可能具有不同的基因表达模式。我们的总体目标是生成小鼠主要且功能重要的颅面缝线的全面基因表达图谱，这将加速我们对人类缝线生物学的理解，并发现其突变可能导致颅缝早闭或颅面骨发育的其他缺陷的候选基因。我们将应用最先进的激光捕获显微切割技术，从正常和颅缝早闭小鼠模型的不同颅面缝线中获取组织，并结合提取的 RNA 的下一代测序 (RNA-Seq)。在目标 1 中，我们将培育具有 Fgfr2 S252W 突变的阿佩尔综合征颅缝早闭的小鼠模型，并使用激光捕获显微切割从 WT 和突变小鼠的 11 条颅面缝线中获取细胞。将培育第二个具有 Twist1 杂合无效突变的 Saethre-Chotzen 综合征小鼠模型，以对两种主要缝合线进行比较。在目标 2 中，我们将从 WT 小鼠的不同缝线亚区域提取 RNA，并进行 RNA 测序以生成一套全面的正常缝线基因表达图谱。在目标 3 中，我们将从 Apert 和 Saethre-Chotzen 综合征小鼠的缝线亚区域中提取 RNA 进行 RNA 测序，并生成与正常基因表达图谱互补的基因表达图谱。这些图谱将允许快速发现缝线中未知表达的基因，揭示缝线之间的共性和差异，这可能提出缝线形成和分化的新假设，对脊椎动物头骨的进化研究具有更广泛的意义，并提供见解进入缝线融合的病理学。