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应用程序包含了一项全面的研究和培训计划 研究嗅觉系统中的离子通道和通道病变。离子通道对于 调节多种细胞类型的兴奋性，包括嗅觉神经元(OSNs)。这些 蛋白质调节离子在细胞膜上的移动。在OSNs离子通道中 负责对气味刺激的细胞去极化，启动一个动作 潜在和突触传递。通道病是人类遗传性疾病中的一类 哪一种离子通道功能被破坏，导致多器官系统的缺陷。中断 已知离子通道可导致癫痫、心律失常、失明、耳聋和 疼痛敏感度的改变。人类患者的通道病可由以下两种因素引起： 离子通道基因的功能和功能增益突变。最近有证据表明， 通道病可能会导致人类嗅觉障碍。中几种不同离子通道的缺失 小鼠模型也会导致嗅觉障碍，这表明嗅觉信号可以在 多个步骤。这项拟议的研究将分析基因疗法纠正疾病的能力 经络病在这些不同的信号传递阶段导致嗅觉功能的缺陷。目标1 这项提议将使用两个在环核苷酸门控中具有靶向缺失的小鼠品系 (CNG)通道亚单位。CNGA2和CNGB1是嗅觉CNG通道的关键亚单位 气味检测所必需的，它们的丢失会导致嗅觉障碍。使用腺病毒，我会传递 将缺失的基因复制到突变的OSN以测试恢复嗅觉的能力 对无嗅觉动物的作用。嗅觉的恢复将被分析为 电生理和行为学方法。目标2将研究基因治疗的能力 纠正经络病引起的钠丢失引起的突触传递障碍 OSN中的通道阿尔法子单元Nav1.7(由SCN9A编码)。使用嗅觉特定的SCN9A NULL 小鼠，我将使用腺病毒在OSN中表达异位SCN9A，并分析其修复情况 突触传递。此外，这个目标将分析功能增益突变的影响 腺病毒介导的突变钠通道在SCN9A细胞中的表达 OSN。这将测试过度兴奋对嗅觉功能的影响。最终目标3将 分析电压门控钙通道(VGCC)α亚基Cav1.3和Cav1.3的作用 Cav2.2，在嗅觉功能方面。VGCC的突变，包括CACNA1D(编码Cav1.3)， 影响感觉功能的潜在经络病症因此是潜在原因 嗅觉障碍的症状。了解这些VGCC在嗅觉系统中的作用将有助于 指导在无嗅觉患者中识别新的突变。加在一起，这3个结果 AIMS将为嗅觉信号的机制和基因能力提供新的见解 纠正离子通道功能缺陷的治疗。这项拟议研究的结果将 对于帮助开发治疗因经络病变而导致的嗅觉障碍的患者具有重要意义。 此外，这些结果可能为开发其他感官的治疗方法提供洞察力。 视力和听力损失等缺陷。