Building the skull - normal and abnormal development

这个项目将专注于颅骨融合，即分离头盖骨的一条或多条缝合线的过早融合。复杂的发育机制网络参与了这一复杂的骨骼系统的形成和维护，各种基因突变可以影响这些过程，导致严重的头骨畸形。牛津大学在这些畸形的外科治疗方面是领先的国家转诊中心，使我们能够研究这些畸形从患者到突变，从老鼠模型到分子发病的整个过程。该项目将利用可用的大量临床样本来探索具体的病因假说。早期的重点将包括对RUNX2基因的研究，该基因编码骨化的主要转录调节因子。作为新的NHS基因医学服务诊断服务研究计划的一部分，很可能有机会询问整个基因组序列。潜在的表观遗传致病机制也将被探索。为了研究病理生理学，精心挑选的突变将在小鼠身上建立模型。我们对颅缝本身在生物学上发生了什么仍然知之甚少：这些结构必须实现微妙的平衡，使缝合边缘的新骨生长，同时确保缝合的中部沿其整个长度保持开放。我们通过绘制从未分化的干细胞到完全形成的成骨细胞的细胞层次结构，比较正常缝合和靶向突变小鼠的结果，来探索缝合是如何工作的。一个特别的焦点将是阐明在实验室可用的ZIC1突变小鼠中引起颅突融合的致病机制。利用CRISPR/Cas9基因组编辑产生更多的靶向小鼠突变体的机会可能来自人类遗传学研究。获得的特殊技能将包括：(I)使用网络资源解释来自人类、小鼠和其他物种的基因组信息；(Ii)实验技术，包括细胞培养、荧光激活细胞分选(FACS)、显微镜、单细胞转录和下一代测序，以及基本的分子生物学方法；(3)小鼠饲养和表型分析方面的培训，获得个人许可证；(4)采用生物信息学方法分析大型数据集，包括基因组测序、单细胞转录组学或表观基因组学(将通过计算生物学中心提供全面支持)。将通过MRC-WIMM DPHIL课程和医学司的技能培训方案提供额外的通用和可转让技能培训。

This project will focus on craniosynostosis, the premature fusion of one or more sutures separating the bones of the skull vault. A complex network of developmental mechanisms is involved in patterning and maintaining this complex system of bones, and a variety of genetic mutations can affect these processes to cause serious skull malformations. Oxford is a leading national referral centre in the surgical treatment of these malformations, enabling us study the entire process by which these arise from patient to mutation, and from mouse model to molecular pathogenesis.The project will exploit the large number of clinical samples available to explore specific hypotheses of causation. An early focus will involve investigation of the gene RUNX2, which encodes a master transcriptional regulator of ossification. There is likely to be the opportunity to interrogate whole genome sequence as part of the research programme of the new NHS Genomic Medicine Service diagnostics service. Potential epigenetic mechanisms of pathogenesis will be explored as well. To investigate pathophysiology, carefully selected mutations will be modelled in mice. We still know very little about what happens biologically in the cranial sutures themselves: these structures must achieve a delicate balancing act of enabling growth of new bone at the margins of the suture, whilst also ensuring that the mid-part of the suture remains open along its entire length. We explore how the suture works by mapping out the cellular hierarchy of activities from undifferentiated stem cell to fully formed osteoblast, comparing results between normal sutures and those from mice with targeted mutations. A specific focus will be to elucidate the pathogenic mechanisms by which craniosynostosis arises in a Zic1-mutated mouse, available in the laboratory. The opportunity to generate additional targeted mouse mutants using CRISPR/Cas9 genome editing may arise out of the human genetics investigations.Specific skills acquired will include: (i) use of web resources to interpret genomic information from human, mouse and other species; (ii) experimental techniques including cell culture, fluorescence-activated cell sorting (FACS), microscopy, single cell transcriptomics and next generation sequencing, in addition to basic molecular biology methodologies; (iii) training in husbandry and phenotypic analysis of mice, with acquisition of a personal license and (iv) bioinformatics approaches to analysis of large datasets including genome sequencing, single cell transcriptomics, or epigenomics (full support will be available through the Centre for Computational Biology). Additional generic and transferable skills training will be provided through the MRC-WIMM DPhil Course and the Medical Sciences Division's Skills Training Programme.