Maintenance and Disassembly of Olfactory Cilia

嗅觉纤毛的维护和拆卸

• 批准号: 10348789
• 负责人: Jeffrey Martens
• 金额: $ 48.2万
• 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 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10348789
• 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; 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 纤毛相关疾病的功能及其改变。嗅觉功能障碍很常见，影响至少 16 仅在美国就有 100 万人。我们的实验室和其他实验室发现嗅觉功能障碍是一种临床表现 一类称为纤毛病的人类遗传性疾病。 Bardet-Biedl 综合征 (BBS) 就是这样一种疾病， 嗅觉感觉神经元（OSN）纤毛形态的改变使细胞对气味没有反应。 尽管已鉴定出许多导致纤毛病的基因，但治愈性疗法（包括嗅觉疗法） 功能障碍）尚未可供患者使用。我们报道了基因替换可以恢复纤毛，从而恢复纤毛 有限数量的分化 OSN 中的感觉输入足以挽救外周气味反应 纤毛病子集的小鼠模型。然而，为了更好地了解嗅觉系统中的纤毛生物学和 为了推进潜在的治疗方法，我们必须定义嗅觉外显率背后的细胞机制 纤毛病。我们还必须确定这些机制在不同纤毛病中是否保守 因此可能适合相同的治疗策略。 OSN 纤毛划分所有必需的 气味检测的信号机制，尽管 OSN 纤毛在生理和环境中都可能丢失 病理条件下，维持这种重要 OSN 细胞完整性的细胞机制 隔间仍然知之甚少。其他细胞和生物体中的纤毛含有多蛋白复合物 碱基，称为过渡区 (TZ)，充当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。该拨款申请将阐明 OSN 纤毛 TZ 的组成和亚细胞组织以及异常蛋白质和脂质易位 进入纤毛有助于纤毛的分解。我们的初步数据表明，细胞内的变化 Ca2+ 通过破坏 TZ 组件来促进纤毛分解。我们假设持续升高 细胞内 Ca2+ 导致 OSN 纤毛 TZ 重塑，从而允许纤毛膜脂质发生变化 分布和肌动蛋白浸润，这两者都是纤毛病中纤毛分解所必需的。所以， 我们提出以下具体目标：（1）确定 OSN 纤毛 TZ 的组成和组织， 纤毛病中伴随拆卸的改变； (2) 确定细胞内Ca2+升高的影响 OSN纤毛拆卸； (3)确定膜PIP2重新分布和F-actin浸润在细胞中的作用 OSN纤毛分解的机制。这项工作的成功完成将提供重要的新见解 进入人类感觉知觉疾病的发病机制，是发展的必要步骤 治疗由纤毛病引起的先天性嗅觉缺失和相关嗅觉功能障碍。