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; 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：嗅觉纤毛的维护和拆卸 该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。嗅觉功能障碍很常见，至少影响16 仅在美国，百万人。我们的实验室和其他人发现嗅觉功能障碍是临床表现 一类称为纤毛病的人遗传疾病。 Bardet-Biedl综合征（BBS）就是一种这样的疾病， 这改变了嗅觉感觉神经元（OSN）的纤毛形态，使细胞对气味无反应。 尽管鉴定出纤毛病的许多基因，但治愈疗法（包括嗅觉 功能障碍）患者尚不可用。我们报道了基因替代品以恢复纤毛，因此 有限数量的分化OSN中的感觉输入足以挽救外围气味反应 纤毛病子集的小鼠模型。但是，要更好地了解嗅觉系统中的纤毛生物学 提前疗法，我们必须定义的细胞机制 纤毛病。我们还必须确定这些机制是否在不同的纤毛病中保守 因此，可能适合相同的治疗策略。 OSN Cilia分隔所有必要的 用于气味检测的信号机械，即使OSN纤毛在生理和 病理条件，维持该必需OSN细胞完整性的细胞机制 隔间仍然很少了解。其他细胞和生物中的纤毛在其上含有多蛋白质复合物 底座称为过渡区（TZ），可作为控制独特蛋白质和脂质的调节门 纤毛的组成。令人惊讶的是，关于OSN中的TZ知之甚少。该赠款申请将阐明 OSN纤毛TZ的组成和亚细胞组织以及异常蛋白和脂质易位如何 进入纤毛助长了纤毛的拆卸。我们的初步数据表明细胞内的改变 Ca2+通过破坏TZ组件来造成纤毛拆卸。我们假设 细胞内Ca2+导致OSN纤毛的TZ重塑，这允许变化纤毛膜脂质 分布和肌动蛋白浸润，这两种纤毛在纤毛病中都是必要的。所以， 我们提出以下具体目的：（1）确定OSN Cilia TZ的组成和组织 纤毛病的拆卸伴随着变化； （2）确定升高的细胞内Ca2+对 OSN纤毛拆卸； （3）确定膜PIP2重新分布和F-肌动蛋白在细胞中的作用 OSN纤毛拆卸的机制。成功完成这项工作将提供关键的新见解 进入人类感觉感知疾病的发病机理，是发展的必要步骤 先天性厌食和相关嗅觉功能障碍的治疗方法是由纤毛病变引起的。