Defining the cellular mechanisms of craniosynostosis in a human induced pluripotent stem cell model of craniofrontonasal syndrome

定义人类颅额鼻综合征诱导多能干细胞模型中颅缝早闭的细胞机制

• 批准号: 9278002
• 负责人: Terren Kathryn Niethamer
• 金额: $ 3.77万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9278002
• 关键词: Actins; Affect; Apoptosis; Behavior; Calvaria; Cell Adhesion; Cell Separation; Cell model; Cells; Clinical; Congenital Abnormality; Craniofacial Abnormalities; Craniosynostosis; Cytoskeleton; Data; Defect; Development; Disease; Dysplasia; Embryo; Ephrin B Receptor; Ephrin-B1; Ephrins; Etiology; Face; Family; Female; Frontal bone structure; Functional disorder; Genes; Human; In Vitro; Individual; Inheritance Patterns; Integral Membrane Protein; Lead; Limb Bud; Link; Maintenance; Mediating; Mesenchymal Differentiation; Mesenchymal Stem Cells; Modeling; Molecular; Mosaicism; Mus; Mutation; Neural Crest; Neuroepithelial Cells; Neurologic; Orbital separation excessive; Osteoblasts; Parietal bone structure; Pathway interactions; Patients; Phenotype; Population; Prediabetes syndrome; Process; Signal Transduction; Signaling Molecule; Sorting - Cell Movement; Structure; Surgical sutures; System; Time; Tissues; X Inactivation; Zebrafish; cell type; coronal synostosis; craniofacial; craniofacial development; craniofrontonasal syndrome; cranium; hindbrain; human disease; induced pluripotent stem cell; loss of function mutation; male; member; migration; mouse model; neural plate; neuroepithelium; novel; osteoblast differentiation; osteogenic; precursor cell; premature; public health relevance; segregation; self organization; skeletal; suture fusion; treatment strategy

DESCRIPTION (provided by applicant): Craniofrontonasal syndrome (CFNS) is an X-linked neurocristopathy characterized by skeletal and neurological anomalies. CFNS is caused by mutations in EFNB1, which encodes EPHRIN-B1, a transmembrane protein and member of the Eph/ephrin family of signaling molecules. Unlike many X-linked conditions, females heterozygous for loss of EFNB1 are more severely affected than hemizygous males. This unique inheritance pattern is due to mosaicism for EFNB1 expression after random X inactivation in heterozygous females. In Efnb1+/- mice, mosaicism leads to active cell sorting between ephrin-B1 expressing and non-expressing cells, resulting in disrupted ephrin-B1 expression boundaries, but the mechanism by which this cell sorting occurs remains incompletely understood. Moreover, cell sorting has not been demonstrated in a human model for CFNS, and it is therefore unknown whether or not cell sorting contributes significantly to disease pathophysiology. One clinically important manifestation of CFNS is coronal craniosynostosis, or premature fusion of the suture between the frontal and parietal bones of the skull. This phenotype is not observed in the Efnb1+/- mouse model, however, presenting an obstacle to understanding its etiology. A human system is therefore needed to understand cellular mechanisms underlying coronal craniosynostosis in CFNS. Human induced pluripotent stem cells (hiPSCs) hold great potential for the study of human disease, as they allow differentiation of patient-specific cells into disease-relevant cell types, which are often difficut or impossible to obtain directly. hiPSCs have been used to model a wide variety of developmental diseases, but they have not been used to model congenital craniofacial anomalies. We have developed a novel hiPSC model for CFNS to study the underlying cellular and molecular etiologies of the disorder and have demonstrated that hiPSC-derived neuroepithelial cells undergo Eph/ephrin-mediated cell sorting. I propose to use our hiPSC model of CFNS to determine the pathways that contribute to Eph/ephrin-mediated cell sorting as well as to study how cell sorting contributes to craniosynostosis in CFNS.

描述（由申请人提供）：颅额鼻综合征 (CFNS) 是一种 X 连锁神经嵴病，其特征为骨骼和神经系统异常。 CFNS 是由 EFNB1 突变引起的，EFNB1 编码 EPHRIN-B1，一种跨膜蛋白，也是 Eph/ephrin 信号分子家族的成员。与许多 X 连锁疾病不同，EFNB1 缺失杂合子的女性比半合子男性受到的影响更严重。这种独特的遗传模式是由于杂合女性中随机 X 失活后 EFNB1 表达的镶嵌现象所致。在 Efnb1+/- 小鼠中，嵌合现象导致 ephrin-B1 表达细胞和非表达细胞之间的主动细胞分选，导致 ephrin-B1 表达边界破坏，但这种细胞分选发生的机制仍不完全清楚。此外，细胞分选尚未在 CFNS 人类模型中得到证实，因此尚不清楚细胞分选是否对疾病病理生理学有显着贡献。 CFNS 的一项重要临床表现是冠状颅缝早闭，或颅骨额骨和顶骨之间缝线的过早融合。然而，在 Efnb1+/- 小鼠模型中没有观察到这种表型，这给理解其病因学带来了障碍。因此，需要一个人体系统来了解 CFNS 冠状颅缝早闭的细胞机制。人类诱导多能干细胞（hiPSC）在人类疾病研究方面具有巨大潜力，因为它们可以将患者特异性细胞分化为疾病相关细胞类型，而这些细胞类型通常很难或不可能直接获得。 hiPSC 已被用于模拟多种发育疾病，但尚未用于模拟先天性颅面异常。我们开发了一种新的 CFNS hiPSC 模型，用于研究该疾病的潜在细胞和分子病因，并证明 hiPSC 衍生的神经上皮细胞经历 Eph/ephrin 介导的细胞分选。我建议使用我们的 CFNS hiPSC 模型来确定有助于 Eph/ephrin 介导的细胞分选的途径，并研究细胞分选如何促进 CFNS 颅缝早闭。