Maintenance and Disassembly of Olfactory Cilia

嗅觉纤毛的维护和拆卸

• 批准号: 10570863
• 负责人: Jeffrey Martens
• 金额: $ 46.89万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10570863
• 关键词: Actins; Affect; Amino Acid Sequence; Animal Model; Anosmia; Applications Grants; Bardet-Biedl Syndrome; Biochemical; Biological; Biological Assay; Biology; Cell Compartmentation; Cells; Cilia; Clinical; Data; Defect; Detection; Disease; Electrophysiology (science); F-Actin; Genes; Genetic; Genetic Diseases; Goals; Hair; Human; Human Genetics; Impairment; Individual; Infiltration; Length; Lipids; Macromolecular Complexes; Maintenance; Measures; Membrane; Membrane Lipids; Molecular; Morphology; Multiprotein Complexes; Nasal cavity; Neurons; Nose; Odors; Olfactory Pathways; Olfactory dysfunction; Organism; Pathogenesis; Pathogenicity; Pathologic; Patients; Penetrance; Perception; Peripheral; Persons; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositols; Phosphoric Monoester Hydrolases; Physiological; Proteins; Regulation; Reporting; Role; Sensory; Signal Transduction; Signaling Protein; Site; Smell Perception; Structure; System; Techniques; Testing; Therapeutic; Wild Type Mouse; Work; base; ciliopathy; congenital anosmia; curative treatments; experience; fluorescence imaging; gene replacement; gene therapy; human model; innovation; insight; mouse model; novel; olfactory sensory neurons; permissiveness; receptive field; response; sensory input; therapy development; virtual

TITLE: Maintenance and Disassembly of Olfactory Cilia The long-term goal of this project is to determine the role of cilia in the regulation and maintenance of olfactory function and their alterations in cilia-related disorders. Olfactory dysfunction is common, affecting at least 16 million people in the U.S. alone. Our lab and others have found olfactory dysfunction to be a clinical manifestation of a class of human genetic disorders termed ciliopathies. Bardet-Biedl syndrome (BBS) is one such disorder, in which the altered cilia morphology of olfactory sensory neurons (OSNs) renders the cells unresponsive to odors. Despite the identification of numerous genes underlying ciliopathies, curative therapies (including for olfactory dysfunctions) are not yet available to patients. We reported that gene replacement to restore cilia and hence sensory input in a limited number of differentiated OSNs was sufficient to rescue peripheral odor responses in mouse models of a subset of ciliopathies. However, to better understand cilia biology in the olfactory system and advance potential therapies, we must define the cellular mechanisms underlying olfactory penetrance of ciliopathies. We must also determine whether these mechanisms are conserved across different ciliopathies and might thus be amenable to the same therapeutic strategies. OSN cilia compartmentalize all of necessary signaling machinery for odor detection and even though OSN cilia can be lost in both physiological and pathological conditions, the cellular mechanisms that maintain the integrity of this essential OSN cell compartment remain poorly understood. Cilia in other cells and organisms contain a multiprotein complex at its base, termed the transition zone (TZ), that functions as a regulatory gate to control the unique protein and lipid composition of cilia. Surprisingly little is known about the TZ in OSNs. This grant application will elucidate the composition and subcellular organization of the OSN cilia TZ and how aberrant protein and lipid translocation into cilia contributes to the disassembly of cilia. Our preliminary data suggest that alterations in intracellular Ca2+ contribute to cilia disassembly by disrupting TZ components. We hypothesize that sustained elevations of intracellular Ca2+ result in TZ remodeling of OSN cilia that is permissive for changes in cilia membrane lipid distribution and actin infiltration, both of which are necessary for cilia disassembly in ciliopathies. Therefore, we propose the following Specific Aims: (1) Determine the composition and organization of the OSN cilia TZ and alterations accompanying disassembly in ciliopathies; (2) Determine the effects of elevated intracellular Ca2+ on OSN cilia disassembly; (3) Determine the role of membrane PIP2 redistribution and F-actin infiltration in cellular mechanisms of OSN cilia disassembly. Successful completion of this work will provide critical new insights into the pathogenesis of human sensory perception diseases and is a necessary step for the development of treatments for congenital anosmia and related olfactory dysfunctions that result from ciliopathy.

标题：嗅纤毛的维持和修复 本项目的长期目标是确定纤毛在嗅觉调节和维持中的作用 功能及其在纤毛相关疾病中的改变。嗅觉功能障碍是常见的，影响至少16 仅在美国就有100万人我们的实验室和其他人发现嗅觉障碍是一种临床表现 一种叫做纤毛病的人类遗传疾病Bardet-Biedl综合征（BBS）就是这样一种疾病， 嗅觉感觉神经元（OSN）纤毛形态的改变使得细胞对气味无反应。 尽管鉴定了许多纤毛病的潜在基因，但治疗性疗法（包括嗅觉疗法）仍然存在。 功能障碍）尚不可用于患者。我们报道了基因替换恢复纤毛， 在有限数量的分化的OSN中，感觉输入足以挽救外周气味反应， 纤毛病变亚组的小鼠模型。然而，为了更好地了解嗅觉系统中的纤毛生物学， 为了推进潜在的治疗方法，我们必须确定嗅觉障碍的细胞机制， 纤毛病变我们还必须确定这些机制是否在不同的纤毛病变中是保守的 因此可能适用于相同的治疗策略。OSN纤毛将所有必要的 气味检测的信号机制，即使OSN纤毛可以在生理和 病理条件下，细胞机制，维持这一基本OSN细胞的完整性 对车厢仍知之甚少。其他细胞和生物体中的纤毛在其末端含有多蛋白复合物。 一个碱基，称为过渡区（TZ），作为一个调节门来控制独特的蛋白质和脂质 纤毛的组成。令人惊讶的是，人们对OSN中的TZ知之甚少。这份补助金申请将阐明 OSN纤毛TZ的组成和亚细胞组织以及异常蛋白质和脂质易位如何 进入纤毛有助于纤毛的分解。我们的初步数据表明，细胞内的改变 Ca ~（2+）通过破坏TZ组分促进纤毛解体。我们假设，持续升高的 细胞内Ca 2+导致OSN纤毛TZ重构，其允许纤毛膜脂质的变化 分布和肌动蛋白浸润，这两者都是纤毛疾病中纤毛解体所必需的。因此，我们认为， 我们提出以下具体目标：（1）确定OSN纤毛TZ的组成和组织， （2）确定细胞内Ca 2+升高对纤毛运动的影响， OSN纤毛解体;（3）确定细胞膜PIP 2重新分布和F-actin浸润在细胞凋亡中的作用。 OSN纤毛解体的机制。这项工作的成功完成将提供重要的新见解 研究人类感觉知觉疾病的发病机制，是发展 治疗先天性嗅觉缺失和相关的嗅觉功能障碍，由纤毛病变引起。