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 习惯性吸烟导致的尼古丁成瘾是美国过早死亡的主要原因 各州每年至少花费 1700 亿美元的医疗保健相关费用。除了让人上瘾之外 尼古丁会引发多种呼吸系统、心血管、胃肠道和免疫疾病， 它的作用不仅反映在吸烟者的大脑中，还反映在吸烟者的身体中。虽然众所周知， 尼古丁的成瘾特性与其对烟碱乙酰胆碱的直接药理作用有关 位于奖赏和动机大脑回路中的受体，我们实验室的证据表明， 尼古丁与其对后脑厌恶回路的作用有关（例如孤束核（NTS）， 脚间核和内侧缰核）在调节尼古丁摄入量方面也发挥着重要作用。 然而，尚不清楚尼古丁是否仅通过直接作用于中枢表达的烟碱来作用于这些回路。 受体或如果尼古丁也通过迷走神经（结节神经节，NG）衍生的感觉输入间接作用，终止 主要在NTS。本次拨款申请中提供的初步数据强烈表明外围设备的作用 尼古丁在控制尼古丁摄入量方面的作用。例如，外周限制性全尼古丁激动剂， 甲基烟碱引起的条件性场所厌恶超过等摩尔剂量尼古丁产生的效果。 此外，外周限制性胆囊收缩素受体 (CCKR) 激动剂 CCK-8 (10 µ*kg-1) 减少 自愿摄入尼古丁，尤其是在致焦虑的尼古丁剂量下。鉴于这些初步数据和 观察到啮齿动物和人类的血浆 CCK 水平受到尼古丁的失调，我假设 肠道神经支配的 NG 神经元中的 CCKR 增强了从外周到 NTS 的厌恶尼古丁信号，从而 调节尼古丁摄入量。我将使用尼古丁静脉注射自我给药小鼠模型来检验这个假设 结合 NG、FosTRAP 小鼠的 CCKR 特异性损伤和化学遗传学。然后我将定义 使用单细胞 RNA 测序研究 NG 对厌恶剂量尼古丁的转录反应。 最后，我将采用体内 CRISPR-Cas9 介导的基因组切割策略来优先敲除 NG 中的尼古丁反应基因并评估对尼古丁摄入的影响。完成这个高度 创新提案将大大提高我在小鼠遗传学、外科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、生物信息学和高级方面的全新培训 基因组编辑技术。它还将为目前关于感官如何进行的稀疏文献做出贡献。 与外周尼古丁作用相关的信息通过迷走神经传递到后脑。定义 这样的机制将使我在这个令人兴奋且快速发展的领域获得成功的独立职业生涯 脑与身体的相互作用。总之，这个培训计划将为我提供概念、技术和 学习经历将成为我过渡到独立研究调查员的跳板。