Changes in Enteric Microbiota and Inflammation with HIV PrEP

项目摘要/摘要 尽管艾滋病毒仍然是一个主要的全球健康问题，但暴露前预防(PrEP)等战略可以 减少新感染的发生率。然而，用于预防艾滋病毒的抗逆转录病毒药物是 与使用它们的个体的肠道炎症增加有关。我们假设它们的广泛活动 同时影响肠道的病毒和细菌群落，引起局部炎症，可能导致上皮 功能障碍，增加微生物移位和全身炎症。 我们提出了一系列元基因组学实验来表征肠道的炎症作用。 病毒和细菌及其随PrEP的变化。对于这项新颖的研究，我们首先必须招募一批 开始使用HIV PrEP的参与者和一组类似的不使用PrEP的对照组参与者。我们会采集血液 并在基线和三个月后采集粪便样本。首先，我们将从粪便样本中鉴定病毒的特征 使用过滤后的病毒颗粒的元基因组测序，并比较内部和之间的变化- PrEP参与者和对照组(目标1)的个体差异。接下来，我们将使用 对包括前驱体在内的粪便细菌种群进行元基因组测序(目标2)。我们会 通过计算预测噬菌体的细菌靶标，然后从 参与者标本。使用这些文化，我们将验证预测的交互并评估 准备扰乱他们。最后，我们将检测粪便和血液中的炎症标志物，并使用病毒和 细菌丰度，以确定哪些微生物与炎症关系最密切(目标3)。这些 实验将识别相互作用并与PrEP相互作用的病毒和细菌，并确定它们的 对宿主炎症的影响。 Maust博士的职业发展计划建立在他的计算生物学背景和实验室专业知识的基础上 通过标记基因测序分析细菌群落，并增加元基因组测序方面的培训， 生物统计学技术，以及病毒和细菌培养的方法。这个K08奖项是一种结构良好的方式 他将最大限度地利用威斯康星大学和西雅图儿童的广泛科学资源来实现独立 作为一名内科科学家，他提出了自己的职业目标，即在微生物影响宿主时了解它们的相互作用 免疫以指导治疗干预。

PROJECT SUMMARY/ABSTRACT Although HIV remains a major global health problem, strategies such as Pre-Exposure Prophylaxis (PrEP) can reduce the incidence of new infections. However, the antiretrovirals used for HIV prevention have been associated with increased enteric inflammation in individuals using them. We hypothesize that their broad activity affects both the viral and bacterial communities of the gut, causing local inflammation which may cause epithelial dysfunction and increase microbial translocation and systemic inflammation. We propose a series of metagenomic experiments to characterize the inflammatory contributions of intestinal viruses and bacteria and their changes with PrEP. For this novel research, we must first enroll a cohort of participants starting HIV PrEP and a similar group of control participants not using PrEP. We will collect blood and stool samples at baseline and after three months. First, we will characterize the virome fromstool specimens using metagenomic sequencing of filtered viral particles and compare the changes of within- and between- individual diversity in the participants on PrEP and those in the control group (aim 1). Next, we will use metagenomic sequencing to characterize stool bacterial populations, including prophages (aim 2). We will computationally predict bacterial targets of bacteriophages, then culture the identified virus-bacterial pairs from participant specimens. Using these cultures, we will validate the predicted interactions and assess the ability of PrEP to disrupt them. Finally, we will measure inflammatory markers in stool and blood and use the viral and bacterial abundances to identify which organisms are most strongly associated with inflammation (aim 3). These experiments will identify viruses and bacteria which interact with each other and with PrEP and define their impact on host inflammation. Dr. Maust’s career development plan builds on his background in computational biology, and lab expertise in analyzing bacterial communities by marker gene sequencing and adds training in metagenomic sequencing, biostatistical techniques, and methods for viral and bacterial culture. This K08 award is a well-structured way for him to make maximal use of UW and Seattle Children’s extensive scientific resources to achieve independence as a physician scientist, advancing his career goal to understand microbial interactions as they impact host immunity to guide therapeutic interventions.