Building the skull - normal and abnormal development

这个项目将集中在颅缝闭合，一个或多个分离颅骨拱顶骨的缝合线的过早融合。一个复杂的发育机制网络参与了这个复杂的骨骼系统的形成和维持，各种基因突变可以影响这些过程，导致严重的颅骨畸形。牛津大学在这些畸形的外科治疗方面是全国领先的转诊中心，使我们能够研究这些畸形从患者到突变，从小鼠模型到分子发病机制的整个过程。该项目将利用大量的临床样本来探索因果关系的具体假设。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。很可能有机会审问全基因组序列，作为新的NHS基因组医学服务诊断服务的研究计划的一部分。并探讨其潜在的表观遗传机制。为了研究病理生理学，精心挑选的突变将在小鼠中建模。我们仍然对颅骨缝合线本身发生的生物学变化知之甚少：这些结构必须达到一种微妙的平衡，既能使缝合线边缘的新骨生长，又能确保缝合线的中间部分沿其整个长度保持开放。我们通过绘制从未分化干细胞到完全形成的成骨细胞的细胞活动等级，比较正常缝合线和靶向突变小鼠缝合线的结果，来探索缝合线是如何工作的。具体的重点将是阐明在实验室中可用的zic1突变小鼠中颅缝闭锁发生的致病机制。利用CRISPR/Cas9基因组编辑产生更多靶向小鼠突变体的机会可能出现在人类遗传学研究中。获得的具体技能将包括：(i)利用网络资源解读人类、小鼠和其他物种的基因组信息；（ii）实验技术，包括细胞培养、荧光活化细胞分选（FACS）、显微镜、单细胞转录组学和下一代测序，以及基本的分子生物学方法；（iii）获得个人执照后进行饲养和小鼠表型分析方面的培训；（iv）利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（计算生物学中心将提供全面支持）。其他通用和可转移技能培训将通过MRC-WIMM博士课程和医学科学司的技能培训计划提供。

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.