Rescue of channelopathy induced anosmias

拯救通道病引起的嗅觉丧失

• 批准号: 8618229
• 负责人: Jeremy McIntyre
• 金额: $ 10.11万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8618229
• 关键词: Action Potentials; Address; Adenoviruses; Affect; Afferent Neurons; Animals; Anosmia; Arrhythmia; Axon; Behavior; Behavioral; Blindness; Calcium Channel; Cell physiology; Cells; Cellular Membrane; Cilia; Collaborations; Complement; Cyclic Nucleotides; Defect; Detection; Development; Disease; Ectopic Expression; Electrophysiology (science); Epilepsy; Esthesia; Exhibits; Food; Functional disorder; Gene Expression; General Population; Genes; Goals; Grant; Hearing; Hereditary Disease; Human; Human Genetics; Individual; Ion Channel; Ions; Knockout Mice; Knowledge; Lead; Link; Martens; Mediating; Messenger RNA; Methods; Modeling; Molecular; Mouse Strains; Movement; Mus; Mutation; Odors; Pain; Patients; Phase; Proteins; Quality of life; Recovery of Function; Reporting; Research; Research Training; Role; Scientist; Sensory; Signal Transduction; Smell Perception; Sodium Channel; Staging; Synaptic Transmission; System; Techniques; Testing; Therapeutic; Therapeutic Intervention; Training; Work; adenoviral-mediated; body system; cell type; ciliopathy; cyclic-nucleotide gated ion channels; deafness; emotional distress; gain of function mutation; gene therapy; hearing impairment; insight; loss of function; mouse model; mutant; neuronal excitability; neurotransmitter release; novel; public health relevance; research study; response; restoration; therapeutic development; therapeutic gene; therapy development; voltage

Project Summary The proposed K99/R00 application incorporates a comprehensive research and training plan for studying ion channels and channelopathies in the olfactory system. Ion channels are critical for regulating excitability in many cell types including olfactory sensory neurons (OSNs). These proteins regulate the movement of ions across the cellular membrane. In OSNs ion channels are responsible for depolarizing the cell in response to odor stimulation, initializing an action potential and synaptic transmission. Channelopathies are a class of human genetic disorders in which ion channel function is disrupted leading to defects in multiple organ systems. Disruptions in ion channels are known to cause epilepsy, cardiac arrhythmias, blindness, deafness and alterations in pain sensitivity. Channelopathies in human patients can result from both loss-of- function and gain-of-function mutations in ion channel genes. It has recently been shown that channelopathies can result in anosmia in humans. Deletions of several different ion channels in mouse models also causes anosmia, indicating that olfactory signaling can be affected at multiple steps. The proposed research will analyze the ability of gene therapy to correct channelopathy induced defects in olfactory function at these different stages in signaling. Aim 1 of this proposal will use two mouse strains with targeted deletions in cyclic nucleotide gated (CNG) channel subunits. CNGA2 and CNGB1 are critical subunits of the olfactory CNG channel necessary for odor detection and their loss leads to anosmia. Using adenovirus, I will deliver functional copies of the missing gene to mutant OSNs to test the ability to restore olfactory function to anosmic animals. Restoration of the sense of smell will be analyzed with electrophysiological and behavioral methods. Aim 2 will investigate the ability of gene therapy to correct channelopathy induced defects in synaptic transmission due to loss of the sodium channel alpha-subunit Nav1.7 (encoded by Scn9a) in OSNs. Using olfactory specific Scn9a null mice, I will use adenovirus to express ectopic Scn9a in OSNs and analyze the restoration of synaptic transmission. In addition this aim will analyze the effects of gain-of-function mutations in Scn9a through adenovirus-mediated expression of identified mutant sodium channels in OSNs. This will test the effect of hyper-excitability on olfactory function. Finally Aim 3 will analyze the role of two voltage gated calcium channel (VGCC) alpha-subunits, Cav1.3 and Cav2.2, in olfactory function. Mutations in VGCCs, including CACNA1D (encoding Cav1.3), underlie channelopathy disorders affecting sensory function and are therefore potential causes of anosmia. An understanding of the function of these VGCCs in the olfactory system will help to direct the identification of novel mutations in anosmic patients. Together the results from these 3 aims will provide new insight into the mechanisms of olfactory signaling and the ability of gene therapy to correct defects in ion channel function. The results from the proposed research will be important for helping to develop therapies for patients with anosmia due to channelopathies. In addition these results may provide insight into developing treatments for other sensory defects such as vision and hearing loss.

项目摘要 拟议的K99/R00应用程序包含了针对的全面研究和培训计划 研究嗅觉系统中的离子通道和通道病。离子通道对于 调节许多细胞类型的兴奋性，包括嗅觉感觉神经元（OSN）。这些 蛋白质调节离子在整个细胞膜上的运动。在OSN离子通道中 负责响应气味刺激的细胞去极化，初始化动作 潜力和突触传播。通道病是一类人遗传疾病 哪个离子通道函数被中断导致多个器官系统中的缺陷。干扰 已知在离子通道中会引起癫痫，心律不齐，失明，耳聋和 疼痛敏感性的改变。人类患者的通道病可能是由于两者丧失的 离子通道基因中的功能和功能突变。最近已经证明 通道病可能会导致人类的厌食症。删除几个不同的离子通道 鼠标模型还引起厌食症，表明嗅觉信号在 多个步骤。拟议的研究将分析基因治疗纠正的能力 通道病在信号传导的这些不同阶段诱导嗅觉函数的缺陷。目标1 该提案将使用两种小鼠菌株在循环核苷酸门中具有靶向缺失 （CNG）通道亚基。 CNGA2和CNGB1是嗅觉CNG通道的关键亚基 气味检测及其损失所必需的导致厌食。使用腺病毒，我会交付 缺失基因到突变OSN的功能副本，以测试恢复嗅觉的能力 对动物的功能。恢复嗅觉将与 电生理和行为方法。 AIM 2将研究基因治疗的能力 由于钠的损失，正确的通道病诱导突触传播中的缺陷 OSN中的通道Alpha-Subunit NAV1.7（由SCN9A编码）。使用嗅觉特异性scn9a null 小鼠，我将使用腺病毒在OSN中表达异位SCN9A并分析恢复 突触传输。此外，此目标将分析功能获得突变的影响 通过腺病毒介导的鉴定突变钠通道的表达在SCN9A中 OSN。这将测试高兴奋性对嗅觉功能的影响。终于瞄准3将 分析两个电压门控钙通道（VGCC）alpha-subunits，Cav1.3和 cav2.2，在嗅觉函数中。 VGCC中的突变，包括cacna1d（编码CAV1.3）， 影响感官功能的基础通道病疾病，因此是潜在的原因 厌食。了解这些VGCC在嗅觉系统中的功能将有助于 指导鉴定异性患者中新型突变。这3的结果在一起 目标将为嗅觉信号的机制和基因的能力提供新的见解 纠正离子通道功能缺陷的治疗。拟议研究的结果将 由于通道病，帮助为厌食症患者开发疗法很重要。 此外，这些结果可能会洞悉开发其他感觉的治疗方法 视力和听力丧失等缺陷。