Investigating microbiota of the gut-brain axis and the impact of cocaine

项目摘要/摘要 可卡因滥用仍然是一种全国性的危机，在美国，吸毒成瘾的代价是 与犯罪活动有关，丧失工作生产力，以及医疗保健，每年总计超过7400亿美元。在……里面 特别是占美国人口13.4%的非裔美国人，他们承担着不成比例的 可卡因的使用和过量。多项研究表明，长期使用可卡因会影响肠道微生物区系， 这反过来会改变下丘脑-脑垂体轴的活动和情绪行为。与可卡因相关的研究 研究表明，滥用药物会引起革兰氏阳性细菌的优先增殖和革兰氏阳性细菌的减少。 阴性类杆菌，结果与与药物成瘾有关的微生物区系一致。积累 证据还表明，肠道微生物会影响肠道的双向交流-- 脑轴，这表明肠道微生物区系对可卡因的关键作用。既然有证据证明这种影响 常见滥用药物对肠道-脑轴、横结肠、远端回肠、基底 神经节，以及垂体和肾上腺可能具有潜在的生物标志物作用，特别是在 过量用药案例，由于可卡因滥用对这些组织造成损害。在这项提案中，我们将解决 可卡因对肠道-脑轴微生物区系的影响。我们的方法将允许评估 利用从可卡因中提取的标本研究肠道-脑轴相关微生物区系中的微生物特征 过量的身体，一种以前的研究没有使用过的方法，但可以提供帮助的信息 确定人们对可卡因的不同反应，使研究人员能够更好地研究 微生物的不良影响。具体地说，我们将测试假设，即存在显著的关联 来自三个南方地区的可卡因过量病例的肠道微生物群和肠道-脑轴之间的关系-阿拉巴马州， 佛罗里达州和田纳西州。为了验证这一假设，我们将(1)确定肠道微生物群，即 与横结肠和远端回肠相关在可卡因使用和致死之间的关系 美国南部地区非裔美国人的药物中毒；(2)确定肠道是否 与基底节、脑垂体腺和肾上腺相关的微生物组介导 南方地区非裔美国人可卡因使用与致命性药物中毒的关系 美国；以及(3)在单盲研究中验证特定微生物生物标志物的数学模型 使用非裔美国人的纸巾来增加信心，并展示可重复性。这些进程将是 评估和验证，为新的诊断和治疗目标提供基础，以防止故意 和无意的可卡因过量，长期目标是恢复肠道微生物群的动态平衡。在……里面 此外，本科生获得了操作苯酚-氯仿的实践经验 提取、聚合酶链式反应和德克萨斯州弗里曼牧场身体农场的研讨会 该项目的大学将为他们提供关于生物医学科学的学术生涯是否 可能是他们想要进一步追求的一条途径。

PROJECT SUMMARY / ABSTRACT Cocaine abuse continues to be a national crisis, with the cost of addiction to illegal drugs in the United States related to criminal activity, lost work productivity, and medical care totaling more than $740 billion per year. In particular, African-Americans, who make up 13.4% of the U.S. population, disproportionately bear the burden of cocaine use and overdoses. Multiple studies have shown that prolonged use of cocaine affects the gut microbiota, which in turn alters hypothalamus-pituitary-axis activity and emotional behavior. Cocaine-related studies have shown that drugs of abuse trigger preferential proliferation of gram-positive Firmicutes and reduction of gram- negative Bacteroidetes, results that are consistent with microbiota associations with drug addiction. Accumulating evidence also shows that gut microorganisms influence the bidirectional communication along the gut- brain axis, which indicates the crucial role of the gut microbiota on cocaine. Since evidence exists for the influence of commonly abused drugs on the microbiota gut-brain axis, the transverse colon, the distal ileum, the basal ganglia, and the pituitary and adrenal glands may have potential roles as biomarkers, specifically in overdose cases, due to the damage to these tissues caused by cocaine abuse. In this proposal, we will address the impact that cocaine has on the microbiota of the gut-brain axis. Our approach will allow for evaluation of microbial signatures in microbiota gut-brain axis-associated tissues using specimens extracted from cocaine overdose cadavers, an approach that previous studies have not used but could provide information that helps determine how people respond differently to cocaine, allowing researchers to better examine the potential adverse effects of microbes. Specifically, we will test the hypothesis that there is a significant association between gut microbiota and the gut-brain axis in cocaine overdose cases from three Southern regions - Alabama, Florida, and Tennessee. To test this hypothesis, we will (1) determine whether the gut microbiome that is associated with the transverse colon and distal ileum mediates the relationship between cocaine use and lethal drug intoxication in African Americans from Southern regions of the United States; (2) determine whether the gut microbiome that is associated with the basal ganglia and the pituitary and adrenal glands mediates the relationship between cocaine use and lethal drug intoxication in African Americans from Southern regions of the United States;; and (3) validate a mathematical model of specific microbial biomarkers in single-blind studies using African-American tissues to increase confidence and demonstrate reproducibility. These processes will be evaluated and validated to provide a basis for new diagnostics as well as therapeutic targets to prevent intentional and unintentional cocaine overdose with the long-term objective of restoring gut microbiome homeostasis. In addition, the hands-on experience that the undergraduate students gain conducting phenol-chloroform extractions, polymerase chain reaction, and the workshop at the Freeman Ranch Body Farm at Texas State University in this project will provide them with insights into whether an academic career in biomedical sciences might be an avenue they want to further pursue.