Nicotine & Nodose: A Neural Basis for Peripheral Control over Nicotine Intake

尼古丁

• 批准号: 10671471
• 负责人: Kevin Braunscheidel
• 金额: $ 6.95万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-06 至 2025-07-05
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10671471
• 关键词: 4-Hydroxy-Tamoxifen; Ablation; Afferent Neurons; Agonist; Applications Grants; Behavior; Bioinformatics; Biological Process; Brain; Brain region; Breeding; C57BL/6 Mouse; CRISPR/Cas technology; Cardiovascular Diseases; Catheters; Cessation of life; Cholecystokinin; Cholecystokinin Receptor; Cigarette Smoker; Clustered Regularly Interspaced Short Palindromic Repeats; Communication; Consummatory Behavior; Data; Development; Disabled Persons; Dissection; Dose; Electric Stimulation; Emerging Technologies; Esthesia; Fiber; Food; Ganglia; Gastrointestinal Diseases; Genes; Genetic; Genetic Transcription; Genomics; Habenula; Habits; Healthcare; Heart; Human; Immune System Diseases; Implant; Infusion procedures; Inhalation; Injections; Intake; Intravenous; Investigation; Laboratories; Learning; Lesion; Life Style; Literature; Lung; Maintenance; Mastication; Medial; Mediating; Mentors; Morbidity - disease rate; Motivation; Mus; Neurons; Nicotine; Nicotine Dependence; Nicotinic Agonists; Nicotinic Receptors; Nodose Ganglion; Nucleus solitarius; Operative Surgical Procedures; Organ; Pattern; Peptides; Peripheral; Pharmaceutical Preparations; Pharmacologic Actions; Plasma; Play; Population; Positioning Attribute; Property; Public Health; Recording of previous events; Relapse; Research; Research Personnel; Respiration Disorders; Rewards; Risk; Rodent; Role; Satiation; Self Administration; Self Assessment; Sensory; Signal Pathway; Signal Transduction; Sincalide; Site; Smoker; System; Technical Expertise; Technology; Testing; Tobacco; Tobacco Dependence; Tobacco smoke; Tobacco use; Training; United States; Up-Regulation; Vagus nerve structure; Virus; addiction; blood-brain barrier crossing; body system; career; cigarette smoking; computer program; cost; diet and exercise; differential expression; experience; ganglion cell; genetic approach; genome editing; hindbrain; in vivo; innovation; interpeduncular nucleus; intravenous administration; knock-down; mind body interaction; mortality; mouse genetics; mouse model; multidisciplinary; nervous system disorder; neural; neural circuit; neuropsychiatric disorder; nicotine self-administration; nicotine use; novel; premature; rapid growth; receptor; response; sensory input; sensory system; single cell sequencing; single-cell RNA sequencing; tobacco control; tobacco smokers; tool; transmission process

ABSTRACT Nicotine addiction in the form of habitual tobacco use is the leading cause of premature death in the United States and costs at least $170 billion in healthcare-related expenses each year. In addition to its addictive qualities, nicotine triggers a plethora of respiratory, cardiovascular, gastrointestinal, and immune disorders, reflecting its actions in not only the brain, but also in the body of smokers. While it is well established that the addictive properties of nicotine are related to its direct pharmacological actions on nicotinic acetylcholine receptors located in reward and motivation brain circuits, evidence from our lab suggests that noxious effects of nicotine related to its actions on hindbrain aversion circuits (e.g. nucleus of the solitary tract (NTS), interpeduncular nucleus, and medial habenula) play a significant role in regulating nicotine intake as well. However, it is unclear if nicotine acts solely on these circuits by direct action on centrally expressed nicotinic receptors or if nicotine also acts indirectly via vagally (nodose ganglia, NG) derived sensory inputs that terminate primarily at the NTS. Preliminary data presented in this grant application strongly suggest a role for peripheral actions of nicotine in controlling nicotine intake. For instance, the peripherally-restricted, full nicotine agonist, methylnicotinium causes a conditioned place aversion beyond that generated by an equimolar dose of nicotine. Further, the peripherally-restricted cholecystokinin receptor (CCKR) agonist, CCK-8 (10 µ*kg-1) decreased volitional nicotine intake, especially at anxiogenic nicotine doses. Given these preliminary data and the observation that plasma CCK levels are dysregulated by nicotine in rodents and humans, I hypothesize that CCKRs in gut-innervating NG neurons potentiate aversive nicotine signals from the periphery to the NTS thereby regulating nicotine intake. I will test this hypothesis using a nicotine intravenous self-administration mouse model in combination with CCKR-specific lesions of the NG, FosTRAP mice, and chemogenetics. I will then define the transcriptional responsiveness of the NG to an aversive dose of nicotine using single cell RNA sequencing. Finally, I will employ an in vivo CRISPR-Cas9-mediated genomic cleavage strategy to knockdown prioritized nicotine-response genes in the NG and assess the consequences on nicotine intake. Completion of this highly innovative proposal will substantially advance my technical skills in mouse genetics, surgery, and computer programing, and provide me with entirely new training in single cell sequencing, bioinformatics, and advanced genome editing technologies. It will also contribute to the currently sparse literature about how sensory information related to nicotine actions in the periphery are transmitted via the vagus to the hindbrain. Defining such a mechanism will position me for a successful independent career in the exciting and rapidly growing field of brain-body interactions. In summation, this training plan will provide me with the conceptual, technical, and learning experiences that will serve as a springboard for my transition to an independent research investigator.

抽象的 以习惯性烟草使用的形式尼古丁成瘾是统一死亡过早死亡的主要原因 州和每年至少要花1700亿美元的医疗费用。除了其添加剂 质量，尼古丁会触发大量呼吸道，心血管，胃肠道和免疫疾病 反映其不仅在大脑，而且在吸烟者体内的行为。虽然已经确定 尼古丁的添加特性与其对烟碱乙酰胆碱的直接药物作用有关 来自奖励和动机脑电路的受体，我们实验室的证据表明，有害影响 尼古丁与其对后脑厌恶电路的作用有关（例如，固体核（NTS）的核 骨间核和内侧Habenula）在调节尼古丁摄入量也起着重要作用。 但是，目前尚不清楚尼古丁是否仅通过对中央表达的烟碱的直接作用才能在这些电路上作用 接收器或尼古丁也通过终止的感官输入（nodose神经节，ng）间接起作用 首先在NTS。本赠款应用程序中提供的初步数据强烈暗示了外围的角色 尼古丁在控制尼古丁摄入方面的作用。例如，外围限制的，全尼古丁激动剂， 甲基二素引起的条件位置的厌恶之外是由等差剂量的尼古丁产生的。 此外，外围限制的胆囊动蛋白受体（CCKR）激动剂，CCK-8（10 µ*kg-1）精制 自愿性尼古丁的摄入量，尤其是在焦虑症尼古丁剂量下。鉴于这些初步数据和 观察到血浆CCK水平在啮齿动物和人类中尼古丁失调，我假设 肠内NG神经元中的CCKR潜在的厌恶尼古丁信号从外围到NTS 调节尼古丁的摄入量。我将使用尼古丁静脉内自我管理鼠标模型检验该假设 结合Ng，Fostrap小鼠和化学遗传学的CCKR特异性病变。然后我将定义 使用单细胞RNA测序，NG对NG厌恶剂量的转录反应能力。 最后，我将采用体内CRISPR-CAS9介导的基因组裂解策略来敲定优先级 NG中的尼古丁反应基因并评估对尼古丁摄入的后果。完成此功能 创新的建议将大大提高我在鼠标遗传学，手术和计算机方面的技术技能 编程，并为我提供单细胞测序，生物信息学和先进的全新培训 基因组编辑技术。它还将有助于当前有关感官的稀疏文献 与外周种中尼古丁作用有关的信息通过迷走神经传播到后脑。定义 这样的机制将使我成为令人兴奋且快速发展的领域的成功独立职业 脑体相互作用。总而言之，该培训计划将为我提供概念，技术和 学习经验将作为我过渡到独立研究调查员的跳板。