Epigenomic labeling of cells that drive drug abuse behavior

驱动药物滥用行为的细胞的表观基因组标记

• 批准号: 10653905
• 负责人: William Russell Renthal
• 金额: $ 53.7万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10653905
• 关键词: Affect; Animal Model; Bar Codes; Behavior; Cells; Cessation of life; Chromatin; Chronic; Development; Drug Prescriptions; Drug abuse; Elements; Enhancers; Environment; Exposure to; Gene Expression; Genes; Genetic; Genomic Segment; Knowledge; Label; Libraries; Life; Maps; Medical Care Costs; Modeling; Morphine; Mus; Nature; Neurons; Nucleus Accumbens; Opioid; Patients; Population; Rapid screening; Refractory; Regulator Genes; Regulatory Element; Relapse; Resolution; Respiratory Failure; Self Administration; Stimulus; Technology; Transgenic Mice; Translating; United States; Viral; Work; addiction; brain reward regions; chronic pain patient; drug abstinence; drug seeking behavior; epigenomics; gene conservation; improved; in vivo; innovation; novel; novel strategies; novel therapeutic intervention; opioid use; opioid use disorder; pain relief; side effect; single nucleus RNA-sequencing

PROJECT SUMMARY In an effort to provide pain relief to tens of millions of patients with chronic pain, opioids are one of the most commonly prescribed medications in the United States. Large segments of the population are thus exposed to the detrimental side effects of opioids, which can be life threatening and include addiction and respiratory failure. Compulsive opioid use despite these negative consequences defines opioid use disorder, a condition that is responsible for nearly 50,000 deaths and $80 billion in medical costs annually. Thus, there is an urgent need for the development of improved treatments for opioid use disorder. One of the greatest challenges in treating opioid use disorder is its chronic nature with patients often relapsing after long periods of drug abstinence. Persistent epigenomic changes in the nucleus accumbens of patients with opioid use disorder are thought to contribute to its chronic, relapsing course. It has remained challenging, however, to translate this knowledge into novel therapeutic approaches because it has not been possible to selectively target the epigenomically-modified neurons involved in drug-seeking behavior without also affecting nearby cells in unrelated circuits. Here we present an innovative approach to label cells that drive opioid-seeking behavior based on their unique epigenomic profile. To do this, we will first map at single-cell resolution, the regions of chromatin that are selectively accessible in mouse nucleus accumbens neurons after morphine self-administration, an established model of opioid use disorder. Some of these genomic regions will act as functional gene regulatory elements that activate gene expression (e.g. gene enhancers) after morphine self-administration. To identify these functional gene enhancers, we will generate an adeno-associated viral library in which each putative element promotes the expression of a unique barcode. We will then use single-nucleus RNA-sequencing to rapidly screen this viral library in vivo for the elements that selectively drive expression of their barcode in the nucleus accumbens neurons that have been epigenomically altered by morphine self-administration. The most selective viral candidate will be used to express inhibitory chemogenetic channels for controlling morphine-seeking behavior. Successful completion of this proposal will establish a fundamentally new approach to selectively label, purify, and control cells that drive opioid-seeking behavior. This approach offers a number of advantages over current state-of-the-art technologies including the ability to label cells involved in drug-seeking behavior without need for transgenic mice or precisely timed conditioned stimuli. By using evolutionarily-conserved gene regulatory elements to drive viral expression, this approach also has the potential to translate to patients with refractory opioid use disorder.

项目摘要 为了减轻数以千万计的慢性疼痛患者，阿片类药物是最多的患者之一 在美国通常开处方药。因此，大部分人口暴露于 阿片类药物的有害副作用，可能是威胁生命的，包括成瘾和呼吸衰竭。 强迫性阿片类药物的使用尽管有这些负面后果定义了阿片类药物的使用障碍，疾病是 每年造成近50,000人死亡和800亿美元的医疗费用。因此，迫切需要 改进治疗阿片类药物使用障碍的发展。 治疗阿片类药物使用障碍最大的挑战之一是其慢性性质，患者经常复发 经过长时间的戒毒。患有 人们认为阿片类药物使用障碍会导致其慢性复发过程。它仍然具有挑战性， 但是，将这些知识转化为新颖的治疗方法，因为不可能 有选择地靶向参与毒品行为的表观遗传学修饰的神经元，而不影响 在无关电路中附近的细胞。 在这里，我们提出了一种创新的方法，用于标签细胞，该细胞根据其独特 表观基因组谱。为此，我们将首先以单细胞分辨率映射，即染色质区域 吗啡自我给药后，有选择地在小鼠核伏伏神经中访问，一个已建立的 阿片类药物使用障碍的模型。其中一些基因组区域将充当功能基因调节元件 在吗啡自助给药后激活基因表达（例如基因增强子）。识别这些 功能性基因增强子，我们将生成一个与腺相关的病毒库，每个假定的元素 促进唯一条形码的表达。然后，我们将使用单核RNA序列进行快速筛选 这个病毒文库在体内的体内，用于选择性地驱动其条形码表达的元素 被吗啡自我给药从表观遗传学上改变的伏隔神经元。最选择性 病毒候选者将用于表达控制吗啡的抑制性化学通道 行为。 该提案的成功完成将建立一种从根本上建立新的方法，以选择性标签，纯化， 以及驱动阿片类药物行为的控制细胞。这种方法比当前具有许多优势 最先进的技术，包括标记参与毒品行为的细胞的能力，而无需 转基因小鼠或精确定时条件刺激。通过使用进化保存的基因调节 驱动病毒表达的元素，这种方法也有可能转化为难治性患者 阿片类药物使用障碍。

项目成果

Epigenomic profiling of mouse nucleus accumbens at single-cell resolution.

单细胞分辨率下小鼠伏隔核的表观基因组分析。

• DOI: 10.1016/j.mcn.2023.103857
• 发表时间: 2023
• 期刊: Molecular and cellular neurosciences
• 作者: Bhatia,Parth;Yang,Lite;Luo,JayXJ;Xu,Mengyi;Renthal,William
• 通讯作者: Renthal,William

Human and mouse trigeminal ganglia cell atlas implicates multiple cell types in migraine.

• DOI: 10.1016/j.neuron.2022.03.003
• 发表时间: 2022-06-01
• 期刊: NEURON
• 影响因子: 16.2
• 作者: Yang, Lite;Xu, Mengyi;Bhuiyan, Shamsuddin A.;Li, Jia;Zhao, Jun;Cohrs, Randall J.;Susterich, Justin T.;Signorelli, Sylvia;Green, Ursula;Stone, James R.;Levy, Dan;Lennerz, Jochen K.;Renthal, William
• 通讯作者: Renthal, William