Utilizing opioid receptor expression to identify the neurons and molecules responsible for opioid respiratory depression and basal breathing.

利用阿片受体表达来识别负责阿片类呼吸抑制和基础呼吸的神经元和分子。

• 批准号: 10701824
• 负责人: Kevin Yackle
• 金额: $ 35.8万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10701824
• 关键词: ARRB2; Acute; Address; Adult; Analgesics; Behavior; Behavioral; Brain; Brain Stem; Breathing; Cardiac pacemaker; Cessation of life; Clinical; Compensation; Data; Future; GTP-Binding Proteins; Generations; Genetic; Goals; In Vitro; Measures; Mediating; Mediator; Medicine; Modeling; Molecular; Motor Neurons; Mus; Neurons; Opioid; Opioid Receptor; Pacemakers; Pain management; Pathway interactions; Periodicity; Persons; Play; Respiratory physiology; Role; Signal Pathway; Signal Transduction; Site; Societies; Synaptic Vesicles; System; Testing; Therapeutic; Time; United States; Ventilatory Depression; Work; beta-arrestin; design; excitatory neuron; experimental study; genetic approach; in vivo; inward rectifier potassium channel; mouse genetics; neural; nodal myocyte; novel; opioid epidemic; opioid misuse; opioid mortality; opioid overdose; opioid use; optogenetics; overdose death; pandemic disease; pharmacologic; preBotzinger complex; presynaptic; prevent; receptor expression; side effect; vesicular release

Between 1999 and 2018 more than 446,000 people died from profoundly slow and shallow breathing after opioid overdose. This number grows at a faster rate each year and in 2019 there were nearly 50,000 overdose deaths. Opioids are perhaps the most effective analgesic, therefore, despite concern, they will persist as a staple therapy in medicine and within our society into the future. Thus, novel solutions to safely use opioids must be developed. But to discover these, we must first understand the key cellular and molecular mechanisms for opioid depression of breathing, known as opioid induced respiratory depression (OIRD). Recently we demonstrated that opioids depress breathing mostly through their action upon µ-opioid receptor (MOR) expressing neurons in the same site where the breathing rhythm is created, the preBötzinger Complex (preBötC). In this proposal, a central goal is to define the key preBötC neural type and MOR signaling pathway(s) that cause OIRD. Results from both aims will have direct clinical and therapeutic impact. Here, we hypothesize that opioids cause OIRD by blocking synaptic vesicle release from just 140 MOR+ excitatory preBötC neurons. Additionally, we expect that these same neurons, despite being less than 10% of preBötC, are specialized for creating the pace of breathing in general, analogous to the cardiac pacemaker cells. If so, these will be the first neurons identified with such an important purpose. This is the second central goal of the proposal. To test these three hypotheses, we have designed a sophisticated intersectional genetic approach to selectively delete MOR in adult mice from either excitatory or inhibitory preBötC neurons or nearby upper airway motor neurons to then compare if and how much OIRD changes. Next, we will use a sensitive in vitro system to uniformly test the necessity of each primary MOR signaling pathway in suppressing preBötC rhythmicity. And last, we will use an intersectional genetic approach to measure changes in breathing after acutely, ectopically silencing the preBötC MOR+ excitatory neurons. The identification of the cellular and molecular mechanisms of OIRD will establish a framework for future studies to develop novel pharmacological approaches to block or rescue it. Moreover, this proposal aims to determine if MOR+ excitatory neurons are pacemakers for breathing. If so, we will have identified, perhaps, some of the most vital neurons in the brain.

在1999年至2018年之间，有446,000多人死于绿op骨后的慢慢和浅呼吸。 过量。这个数字每年的增长速度更快，2019年有近50,000人过量死亡。 因此，阿片类药物可能是最有效的镇痛药，因此，目的地关心，它们将继续作为主食治疗 在医学和我们的社会中进入未来。这是必须开发安全使用阿片类药物的新颖解决方案。 但是要发现这些，我们必须首先了解阿片类药物抑郁的关键细胞和分子机制 呼吸，称为阿片类药物诱导的呼吸抑郁症（OIRD）。 最近，我们证明阿片类药物主要通过其对阿片类药物接收器的作用降低呼吸 （MOR）在产生呼吸节奏的同一位置表达神经元，Prebötzinger综合体 （prebötc）。在此提案中，一个核心目标是定义关键的PreBötc神经类型和MOR信号通路 这会导致oird。两个目标的结果将具有直接的临床和治疗影响。在这里，我们假设 OioID通过阻止仅140个MOR+兴奋性Prebötc神经元的突触囊泡释放而引起的。 此外，我们期望这些相同的神经元（二p岩少于prebötc的10％）专门用于 通常，呼吸速度通常类似于心脏起搏器细胞。如果是这样，这些将是第一个 神经元以如此重要的目的确定。这是该提议的第二个中心目标。测试这些 三个假设，我们设计了一种复杂的交叉遗传方法来选择性删除MOR 在兴奋或抑制性prebötc神经元或附近气道运动神经元的成年小鼠中 比较是否以及多少变化。接下来，我们将使用敏感的体外系统统一测试 每种主要MOR信号通路的必要性在抑制Prebötc节奏性中。最后，我们将使用 急性，生态沉默的呼吸变化的遗传学方法，使呼吸器变化对prebötc进行沉默 MOR+兴奋性神经元。 OIRD的细胞和分子机制的鉴定将建立一个未来研究的框架 开发新颖的药理学方法来阻止或营救它。此外，该建议旨在确定是否是否 MOR+兴奋性神经元是呼吸的起搏器。如果是这样，我们也许已经确定了最多的 大脑中的重要神经元。