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和骨骼的遗传基础 中枢神经系统畸形。我们在过去几年中在人类和小鼠遗传学方面所做的努力 一直将我们引向初级纤毛作为人类信号传递的关键枢纽 健康和疾病。纤毛疾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 (除其他条件外)。从文献中可以清楚地看出，修改基因座是至关重要的 在了解人类疾病的许多方面，但纤毛疾病尤其如此。 这项建议的重点主要是初级纤毛基因四肽重复结构域。 21b(Ttc21b)。Ttc21b纯合子小鼠突变株本身有几个显著的特征，但 我们的初步数据和其他人的工作清楚地表明，TTC21B是人类的中枢 纤毛病变网。我们从人类遗传学中选取了四个候选交互作用 在小鼠体内重建了它们。这四个基因都与Ttc21b相互作用，但细胞和机制 由此产生的表型尚未阐明。我们还确认了多部小说 ENU诱变组合的互作位点及遗传的QTL分析 小头症表型严重程度的背景效应。因此，我们有重要的 并确定了知识中的四个关键差距，我们将通过以下方式解决 Mira奖的支持：1)Ttc21b是如何对器官产生这种组织特异性作用的？ 生理学和发育信号，2)什么是遗传相互作用和修饰物 Ttc21b改变了这些纤毛病变的表型，3)Ttc21b的细胞功能是什么 主纤毛；4)Ttc21b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主题。许多. 纤毛基因被确认在初级纤毛内具有作用，但在外部的任何功能 纤毛还没有被阐明。确定这些角色将具有非常重要的意义 对赛场的影响。根据初步数据，我们最喜欢的假设是Ttc21b具有 在神经元运输中的重要作用。很明显，对人类疾病的更好理解 将需要修改基因座和潜在机制的知识(S)。Ttc21b的时机已经成熟 探索是纤毛疾病遗传网络的重要组成部分。一种鼠标组合 受人类基因组学启发的胚胎学和细胞生物学是一个理想的切入点。我们的工作很可能是 不仅有助于了解纤毛病，而且作为一名将军指明了前进的方向 在许多不同的病理生理环境下的实验范式。

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.