Molecular Analysis of primary cilia proteins in human development

我实验室的研究目标是研究人类颅面的遗传基础， CNS畸形。过去几年我们在人类和小鼠遗传学方面的努力 不断地引导我们将初级纤毛作为人类信号传递的关键枢纽， 健康和疾病。纤毛病是与严重的先天性 畸形以及非致命性颅面畸形、智力残疾和肥胖 （其他条件）。从文献中可以清楚地看出，修饰基因座是至关重要的 这是理解人类疾病的重要组成部分，但对纤毛病尤其如此。 这项建议的重点主要是在初级纤毛基因tetratricopeptide重复结构域 21B（Ttc21b）。ttc 21 b纯合突变小鼠自身具有几个显著特征， 我们的初步数据和其他人的工作清楚地表明，TTC 21 B是人类的一个中心， 纤毛病变网络我们已经从人类遗传学中提取了四种候选相互作用， 在老鼠身上重现。所有四个基因都与Ttc 21 b相互作用，但Ttc 21 b的细胞和机制不清楚。 所得到的表型尚未阐明。我们还发现了多个 相互作用基因座与ENU诱变和遗传的QTL分析的组合 对小头畸形表型严重程度的背景影响。因此，我们有重要的 在该领域的经验，并确定了四个关键的知识差距，我们将解决与 这项MIRA奖的支持：1）Ttc 21 b如何对器官产生组织特异性影响 生理学和发育信号，2）什么是遗传相互作用和修饰剂， Ttc 21 b改变这些纤毛病变表型，3）Ttc 21 b在其中的细胞功能是什么 初级纤毛; 4）Ttc 21 b在纤毛外的作用。我们将结合使用 遗传学，分子胚胎学，细胞生物学和生物化学来解决这些问题。许多 纤毛基因被鉴定为在初级纤毛内起作用，但在初级纤毛外的任何功能， 纤毛尚未阐明。确定这些作用将具有非常重要的意义。 对场的影响。基于初步数据，我们最喜欢的假设是Ttc 21 b具有 在神经元运输中的重要作用。很明显，更好地了解人类疾病 将需要修饰基因座和潜在机制的知识。ttc 21 b是成熟的， 探索作为一个重要组成部分，在纤毛遗传网络。鼠标组合 受人类基因组学启发的胚胎学和细胞生物学是理想的切入点。我们的工作很可能 不仅有助于了解纤毛病变，而且作为一个一般的前进方向， 实验范式的一些不同的病理生理背景。

The goal of the research in my laboratory is to study the genetic basis of human craniofacial and CNS malformations. Our efforts in both human and mouse genetics over the past several years have continually directed us towards the primary cilium as a critical hub in signaling for human health and disease. Ciliopathies are diseases associated with both severe congenital malformations as well as nonlethal craniofacial dysmorphology, intellectual disability and obesity (among other conditions). It is clear from the literature that modifying loci are crucial components in understanding much of human disease, but is especially true of the ciliopathies. The focus of this proposal is largely on the primary cilia gene tetratricopeptide repeat domain 21B (Ttc21b). Ttc21b homozygous mouse mutants have several striking features on their own but our preliminary data and the work of others clearly show that TTC21B is a hub in a human ciliopathy network. We have taken four of these candidate interactions from human genetics and recreated them in mouse. All four genes interact with Ttc21b but the cellular and mechanisms of the resulting phenotypes are not yet elucidated. We have also identified multiple novel interacting loci with a combination of ENU mutagenesis and a QTL analysis of the genetic background effects on the severity of the microcephaly phenotype. Thus, we have significant experience in the field and have identified four crucial gaps in knowledge we will address with the support of this MIRA award: 1) How does Ttc21b have such tissue specific effects on organ physiology and developmental signaling, 2) What are the genetic interactors and modifiers of Ttc21b which alter these ciliopathy phenotypes, 3) what is the cellular function of Ttc21b inside the primary cilium, 4) what is the role of Ttc21b outside the cilium. We will use a combination of genetics, molecular embryology, cell biology and biochemistry to address these topics. Many of the ciliary genes are identified to have roles within the primary cilium, but any function outside the cilium has not been elucidated. Identification of such roles would have very a significant effect on the field. Our favorite hypothesis based on preliminary data is that Ttc21b has significant roles in neuronal trafficking. It is clear that a better understanding of human disease will require knowledge of modifying loci and the underlying mechanism(s). Ttc21b is ripe for exploration as a crucial component in a ciliopathy genetic network. A combination of mouse embryology and cell biology inspired by human genomics is an ideal entry point. Our work is likely to not only contribute to knowledge about ciliopathies but point the way forward as a general experimental paradigm for a number of different pathophysiological contexts.