Therapeutic rescue of an olfactory ciliopathy

描述（由申请人提供）：拟议的NRSA博士后包括一个全面的研究和培训计划，研究嗅觉系统中的蛋白质运输和纤毛功能。纤毛是进化上保守的细胞器，在感知外界刺激和维持细胞稳态方面发挥重要作用。纤毛病是一类新兴的人类遗传疾病，其中纤毛功能被破坏导致各种各样的组织缺陷。人类患者的纤毛病可由纤毛相关基因的点突变或全部缺失引起。受这些纤毛病影响的系统之一是嗅觉系统，导致嗅觉能力的减少或丧失。单个嗅觉感觉神经元（OSNs）包含超过20个纤毛，这些纤毛投射到嗅觉粘膜并负责检测气味。由于嗅觉纤毛缺乏蛋白质合成成分，嗅觉信号蛋白必须通过特定的机制正确靶向并在整个纤毛中移动。蛋白质通过鞭毛内运输（IFT）的过程在纤毛中移动是公认的。IFT在嗅纤毛发育、维持和功能中的作用在很大程度上是未知的。本提案的目的1将使用突变小鼠品系ORPK，其中纤毛的维持和功能被改变。ORPK小鼠无法探测到气味，因为嗅觉纤毛大部分缺失。利用腺病毒，我将把突变基因IFT88的功能拷贝传递给osn，以恢复嗅觉纤毛和重要的嗅觉功能。此外，这一目标将集中在改善基因递送到osn的方法上，包括使用非病毒方法。目的2将研究嗅觉特异性IFT88基因敲除中的纤毛发育和嗅觉功能。突变和缺失都是纤毛病的基础；然而，这些变化引起的表型可能存在差异。在目标2中，我将研究嗅觉特异性敲除中嗅觉纤毛发生的变化，并测试嗅觉功能。最后，我将测试腺病毒基因传递方法是否可以恢复IFT88缺失引起的缺陷。这些研究结果将为IFT在嗅纤毛发育和维持中的作用提供新的见解。重要的是，该项目的结果将有助于开发嗅觉纤毛病患者的治疗方法，以及其他类型的嗅觉功能障碍以及受纤毛病影响的其他器官系统的潜在治疗方法。

DESCRIPTION (provided by applicant): The proposed postdoctoral NRSA incorporates a comprehensive research and training plan for the study of protein trafficking and cilia function in the olfactory system. Cilia are evolutionarily conserved organelles that play important roles in both sensing external stimuli and maintaining cellular homeostasis. Ciliopathies are an emerging class of human genetic disorders in which cilia function is disrupted leading to a wide variety of tissue defects. Ciliopathies in human patients can be caused by point mutations or entire deletions of cilia related genes. One of the systems that are affected by these ciliopathies is the olfactory system, leading to a reduction or loss in the ability to smell. Individual olfactory sensory neurons (OSNs) contain over 20 cilia that project into the olfactory mucosa and are responsible for detecting odorants. Since olfactory cilia lack protein synthesis components, olfactory signaling proteins must be correctly targeted to and moved throughout the cilia by specific mechanisms. It is well established that proteins move throughout cilia by the process of Intraflagellar Transport (IFT). The role of IFT in olfactory cilia development, maintenance and function is largely unknown. Aim 1 of this proposal will use a mutant mouse strain, ORPK, in which cilia maintenance and function is altered. ORPK mice are unable to detect odorants as the olfactory cilia are largely absent. Using adenovirus, I will deliver a function copy of the mutated gene, IFT88, to OSNs in order to restore olfactory cilia and importantly olfactory function. Additionally this aim will concentrate on ways to improve gene delivery to OSNs, including the use of non-viral methods. Aim 2 will investigate cilia development and olfactory function in an olfactory specific IFT88 knockout. Both mutations and deletions underlie ciliopathies; however there may be differences in the phenotypes caused by these changes. In aim 2 I will investigate changes in olfactory ciliogenesis in the olfactory specific knockout as well as testing olfactory function. Finally I will test whether adenoviral gene delivery methods can restore defects caused by IFT88 deletion. The results from these studies will provide new insight into the roles of IFT in olfactory cilia development and maintenance. Importantly the results from this project will help in the development of therapies for patients with olfactory ciliopathies, and potential therapies for other types of olfactory dysfunction as well other organ systems affected by ciliopathies. PUBLIC HEALTH RELEVANCE: Defects in cilia function underlie an emerging class of human genetic diseases, termed ciliopathies, many of which result in the loss of smell. My research aims will help to identify mechanisms that regulate olfactory cilia maintenance and how these are disrupted in disease states. The ultimate goal of this application is to demonstrate functional recovery of olfactory function in a ciliopathy model to further the development of treatments for human ciliopathies.