SINGLE SUTURE CRANIOSYNOSTOSIS: CANDIDATE GENE AND PATHWAY DISCOVERY

单缝颅缝早闭：候选基因和通路的发现

基本信息

批准号：
8824918
负责人：
MICHAEL L CUNNINGHAM
金额：
$ 74.96万
依托单位：
SEATTLE CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-03-25 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8824918
关键词：
Alkaline Phosphatase Animal Model Biological Response Modifier Therapy Biomechanics Breeding Calvaria Candidate Disease Gene Cell Death Cells Child Code Collaborations Congenital Abnormality Congenital abnormal Synostosis Counseling Coupled Craniosynostosis DNA Resequencing Data Development Diagnosis Differentiation and Growth Disease Engineering Environment Experimental Models Extracellular Matrix Family Focal Adhesions Gene Expression Genes Genetic Models Genetic Variation Genome Genomics Health Human IGF1 gene IGF1R gene Individual Interdisciplinary Study Measures Mechanics Medicine Modeling Molecular Molecular Diagnostic Testing Molecular Genetics Morbidity - disease rate Mus Mutant Strains Mice Mutation Nucleic Acid Regulatory Sequences Osteoblasts Pathogenesis Pathologic Pathway interactions Patients Phenotype Polygenic Traits Primary Prevention Property Recruitment Activity Research Resource Development Resources Science Signal Pathway Somatomedins Subgroup Surgical sutures TWIST1 gene Techniques Testing Traction Universities Variant Washington Work base bone clinical care cohort comparative genomic hybridization craniofacial cranium design developmental genetics gain of function gain of function mutation genome sequencing genomic variation improved in vitro Assay loss of function mutation mineralization mortality mouse model network models novel osteoblast differentiation premature suture fusion transcriptome sequencing transcriptomics

项目摘要

DESCRIPTION (provided by applicant): Craniosynostosis is the pathologic fusion of the sutures of the calvaria. It is associated with significant morbidity, occasional mortality, and carries a considerable financial burden. Using a combination of candidate gene resequencing and comparative genome hybridization TWIST1 loss-of-function mutations and IGF1R and RUNX2 gain-of-function mutations have been identified. Expression array analysis coupled with network modeling has identified activation of an IGF1-RUNX2 pathway as a potential cause of craniosynostosis in a large subgroup of cases. The identification of a biologically based subgroup is a critical advance toward understanding the cause of synostosis as it affords an ability to focus research efforts on a phenotypically similar cohort of cases. This competitive renewal proposes the use of genomics, network modeling, animal models and novel cell biologic approaches to reveal genetic and developmental pathways which, when disrupted, result in premature calvarial fusion. New (Igf1rGOF) and existing (Igf1GOF, Gsk3bLOF, Twist1LOF) mouse resources will be used to model the polygenic inheritance of this disorder. Through interdisciplinary research efforts, three independent yet highly integrated aims will test the hypothesis that a subset of children with isolated single suture craniosynostosis has identifiable genetic variation that results in enhanced calvarial osteoblast differentiation throug activation of an IGF1-RUNX2 pathway. Specific Aim 1 will identify changes in the cellular phenotype of osteoblasts demonstrating activation of the IGF1-RUNX2 pathway. We will utilize measures of osteoblast growth and differentiation as well as measures of cellular biomechanics to refine the biologic phenotype of our original cohort. Specific Aim 2 will develop and characterize an inducible Igf1rR407H mouse model of the human IGF1RR406H gain-of-function mutation.11 We will breed and phenotype Igf1rR407H compound heterozygous mice using existing mutant mouse resources (Igf1GOF, Gsk3bLOF, Twist1LOF) to develop models for the polygenic inheritance of SSC in humans. Specific Aim 3 will determine the contribution of genomic variation in the development of craniosynostosis among cases in the IGF1/RUNX2 subgroup. We will use transcriptome sequence data from the original cohort (N=211) and whole genome sequence data from the IGF1/RUNX2 subgroup cases (N=48) to refine the pathway and identify correlates between alteration in gene expression, coding variants and regulatory region variation. We will recruit a new SSC cohort to validate the transcriptomic and genomic variation identified. Major gaps exist in the diagnosis and management of isolated craniosynostosis including the lack of molecular diagnostic testing, adequate family counseling, and biologic therapies to reduce patient morbidity. There is an incomplete understanding of the causes of craniosynostosis and we lack experimental models. The development of these resources will improve clinical care, design biologically based therapies, and pursue primary prevention.

描述（由申请人提供）：颅缝早闭是颅骨缝线的病理性融合。它与显着的发病率、偶发的死亡有关，并带来相当大的经济负担。通过结合候选基因重测序和比较基因组杂交，已鉴定出 TWIST1 功能丧失突变以及 IGF1R 和 RUNX2 功能获得突变。表达阵列分析与网络建模相结合，已确定 IGF1-RUNX2 通路的激活是大量病例亚组中颅缝早闭的潜在原因。基于生物学的亚组的识别是理解骨联原因的关键进步，因为它提供了将研究工作集中在表型相似的病例组上的能力。这种竞争性更新建议使用基因组学、网络模型、动物模型和新颖的细胞生物学方法来揭示遗传和发育途径，这些途径一旦被破坏，就会导致颅骨过早融合。新的 (Igf1rGOF) 和现有的 (Igf1GOF、Gsk3bLOF、Twist1LOF) 小鼠资源将用于模拟这种疾病的多基因遗传。通过跨学科研究工作，三个独立但高度整合的目标将检验以下假设：患有孤立性单缝颅缝早闭的儿童子集具有可识别的遗传变异，通过激活 IGF1-RUNX2 途径导致颅骨成骨细胞分化增强。具体目标 1 将识别成骨细胞细胞表型的变化，证明 IGF1-RUNX2 通路的激活。我们将利用成骨细胞生长和分化的测量以及细胞生物力学的测量来完善我们原始队列的生物表型。具体目标 2 将开发并表征人类 IGF1RR406H 功能获得性突变的可诱导 Igf1rR407H 小鼠模型。 11 我们将使用现有的突变小鼠资源（Igf1GOF、Gsk3bLOF、Twist1LOF）培育 Igf1rR407H 复合杂合小鼠并进行表型分析，以开发模型人类SSC的多基因遗传。具体目标 3 将确定基因组变异在 IGF1/RUNX2 亚组病例中颅缝早闭发生中的作用。我们将使用原始队列 (N=211) 的转录组序列数据和 IGF1/RUNX2 亚组病例 (N=48) 的全基因组序列数据来完善途径并确定基因表达、编码变异和调控区域改变之间的相关性变化。我们将招募一个新的 SSC 队列来验证已识别的转录组和基因组变异。孤立性颅缝早闭的诊断和治疗方面存在重大差距，包括缺乏分子诊断测试、充分的家庭咨询和降低患者发病率的生物疗法。对颅缝早闭的原因尚不完全了解，而且我们缺乏实验模型。这些资源的开发将改善临床护理、设计基于生物学的疗法并进行初级预防。