Dissecting the role of the gut microbiome on striatal chromatin conformation and drug-seeking behavior

剖析肠道微生物组对纹状体染色质构象和药物寻求行为的作用

• 批准号: 10319289
• 负责人: Katherine Meckel
• 金额: $ 4.54万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10319289
• 关键词: Abstinence; Acute; Affect; Animal Model; Animals; Antibiotics; Bacteria; Behavior; Behavioral; Biological Assay; Blood - brain barrier anatomy; Brain; Chromatin; Chromatin Structure; Chronic; Cocaine; Complement; Complex; Corpus striatum structure; Coupled; Cues; DNA Binding; Data; Development; Disease; Drug Controls; Drug Modelings; Drug usage; Epigenetic Process; Exhibits; Exposure to; FDA approved; Faculty; Fellowship; Fiber; Fostering; Future; Gastrointestinal tract structure; Gene Expression; Genes; Genetic Transcription; Goals; Health; Histone Acetylation; Homeostasis; Image; Imaging Techniques; Immune system; Injections; Intravenous; Laboratories; Learning; Long-Term Depression; Long-Term Potentiation; Mental disorders; Mentors; Modeling; Molecular; Molecular Conformation; Morbidity - disease rate; Mus; Neurons; Nuclear; Nucleus Accumbens; Overdose; Pathogenesis; Pathologic; Patients; Pattern; Pharmaceutical Preparations; Pharmacotherapy; Phase; Photometry; Play; Population; Positioning Attribute; Public Health; Publishing; Regulation; Regulator Genes; Relapse; Research; Research Personnel; Rewards; Rodent Model; Role; Signal Pathway; Signal Transduction; Societies; Sorting - Cell Movement; Substance Use Disorder; Synapses; Synaptic plasticity; Techniques; Testing; Training; Transcriptional Regulation; Transposase; Vagus nerve structure; Withdrawal; Work; XCL1 gene; brain behavior; cell type; cocaine exposure; cocaine self-administration; cocaine use; cost; disability; drug craving; drug of abuse; drug reward; drug seeking behavior; epigenome; experimental study; gut microbiome; gut microbiota; gut-brain axis; human subject; in vivo calcium imaging; insight; loved ones; microbial; microbiome; mortality; mouse model; neurotransmission; novel; patient population; post-doctoral training; pre-doctoral; prevent; psychostimulant; response; skills; stimulant use; stimulant use disorder; tenure track; therapeutic target; transcriptome sequencing; transcriptomics

Project Summary Pathological substance use disorders are devastating psychiatric illnesses characterized by patterns of escalating and out of control drug use. These disorders account for profound disability, morbidity and mortality among patients, while also inflicting incalculable costs on their loved ones and society at large. Pathological use of stimulants, including cocaine, accounts for a significant proportion of morbidity and fatal overdose nationwide. Despite tremendous advances in elucidating the molecular changes underlying stimulant use disorders, there are no FDA-approved pharmacotherapies for their treatment. The largest obstacle to overcome in treating patients with psychostimulant use disorder is preventing relapse. In recent years, there has been a growing understanding that the resident population of bacteria in the gastrointestinal tract – collectively referred to as the gut microbiome – plays a critical role in regulating brain function and behavior. Extensive research now demonstrates that changes to the gut microbiome can support healthy brain function and also drive the development of pathological states. In our own lab, we have previously published that acute depletion of the microbiome can enhance the rewarding effects of cocaine and alter gene expression changes in the brain. In this proposal, I will investigate how antibiotic-induced depletion of the microbiome affects the persistence of drug-seeking behaviors in a mouse model of relapse, taking an integrative multi-level approach to examine changes in striatal reward centers at the epigenetic, transcriptional, and circuit levels. Preliminary data in Aim 1 (F99) demonstrate that microbiome-depleted mice exhibit increased cocaine-seeking behaviors following a period of prolonged abstinence. Full transcriptomic RNA-sequencing analysis from the nucleus accumbens demonstrates that these animals exhibit marked changes in gene expression in important gene networks known to affect DNA binding, synaptic plasticity, and behavior. Additional analyses demonstrate decreased permissive histone acetylation in microbiome-depleted mice following cocaine administration, suggesting that the microbiome affects chromatin structure. Aim 2 (F99) will delve more deeply into the microbial regulation of the epigenome in response to drugs of abuse by examining chromatin conformation in specific populations of striatal neurons. This will be accomplished via training in and application of Fluorescent- Activated Nuclear Sorting (FANS) and Assay for Transposase-Accessible Chromatin sequencing (ATAC-seq). These studies will be coupled with behavioral analyses to provide mechanistic insight into epigenetic remodeling underlying drug-seeking behaviors. For Aim 3 (K00) I will identify a postdoctoral mentor to support my professional development as well as my research examining the effects of the microbiome on firing activity in striatal neuronal subpopulations using in vivo calcium imaging techniques. These studies will provide important mechanistic insight into gut-brain signaling in a model of cocaine use disorder while simultaneously fostering my development into a successful independent investigator ready for a tenure-track faculty position.

项目摘要病理性物质使用障碍是一种破坏性的精神疾病，其特征是 毒品使用不断升级和失控的模式。这些障碍导致严重的残疾、发病率 这不仅会降低患者的死亡率，同时也会给他们的亲人和整个社会带来无法估量的代价。 病理性使用兴奋剂，包括可卡因，占发病率和致命性的很大比例。 全国范围内吸毒过量。尽管在阐明兴奋剂潜在的分子变化方面取得了巨大进展 使用障碍，目前还没有FDA批准的药物疗法来治疗他们。最大的障碍是 在治疗精神刺激剂使用障碍的患者中克服了这一点，防止了复发。近年来，有 越来越多的人认识到，胃肠道中的常驻细菌种群- 统称为肠道微生物组-在调节大脑功能和行为方面发挥着关键作用。 广泛的研究表明，肠道微生物群的改变可以支持健康的大脑功能 也推动了病理状态的发展。在我们自己的实验室里，我们之前已经发表了这种急性 微生物组的耗尽可以增强可卡因的奖赏效应并改变基因表达的变化 在大脑里。在这项提案中，我将研究抗生素导致的微生物群枯竭如何影响 多层次综合研究复吸小鼠模型中药物寻找行为的持续性 研究纹状体奖赏中心在表观遗传、转录和回路水平上的变化。初步 Aim 1(F99)中的数据表明，微生物组耗尽的小鼠表现出更多的可卡因寻找行为 在一段长时间的禁欲之后。细胞核的全转录RNA测序分析 伏隔菌表明，这些动物在重要基因的基因表达方面表现出显著的变化。 已知的影响DNA结合、突触可塑性和行为的网络。其他分析表明 在给予可卡因后，微生物组耗竭小鼠中允许的组蛋白乙酰化减少， 这表明微生物群影响染色质结构。《目标2》(F99)将更深入地探讨 通过检测染色质构象对表观基因组的微生物调节对滥用药物的反应 纹状体神经元的特定群体。这将通过培训和应用荧光技术来实现- 激活核分类(FANS)和转座酶可及染色质测序分析(ATAC-SEQ)。 这些研究将与行为分析相结合，以提供对表观遗传学的机制洞察 重塑潜在的毒品寻觅行为。对于目标3(K00)，我将指定一位博士后导师来支持 我的专业发展以及我对微生物群对激发活动影响的研究 体内钙成像技术在纹状体神经元亚群中的应用。这些研究将提供 在可卡因使用障碍模型中对肠脑信号的重要机械洞察力 将我培养成一名成功的独立调查员，为终身教职做好准备。