Changes in Enteric Microbiota and Inflammation with HIV PrEP

项目总结/摘要 虽然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）等策略可以 减少新感染的发生。然而，用于预防艾滋病毒的抗逆转录病毒药物一直是 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 影响肠道的病毒和细菌群落，引起局部炎症， 功能障碍并增加微生物易位和全身炎症。 我们提出了一系列的宏基因组实验来表征肠道炎症的贡献， 病毒和细菌及其与PrEP的变化。对于这项新的研究，我们必须首先招募一个队列， 开始HIV PrEP的参与者和不使用PrEP的类似对照组参与者。我们将收集血液 以及基线和三个月后的粪便样本。首先，我们将从粪便标本中鉴定病毒组， 使用过滤的病毒颗粒的宏基因组测序，并比较 PrEP参与者和对照组参与者的个体多样性（目标1）。接下来，我们将使用 宏基因组测序以表征粪便细菌群体，包括前噬菌体（aim 2）。我们将 通过计算预测噬菌体的细菌靶标，然后培养鉴定的病毒-细菌对， 参与者标本。使用这些培养物，我们将验证预测的相互作用，并评估 PrEP来扰乱他们。最后，我们将测量粪便和血液中的炎症标志物，并使用病毒和 细菌丰度，以确定哪些生物体与炎症最密切相关（目标3）。这些 实验将确定病毒和细菌相互作用，并与PrEP和定义他们的 影响宿主炎症。 博士Maust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，并增加宏基因组测序的培训， 生物统计学技术，以及病毒和细菌培养方法。这个K 08奖项是一个结构良好的方式， 他最大限度地利用华盛顿大学和西雅图儿童广泛的科学资源，以实现独立 作为一名医生科学家，他的职业目标是了解微生物在影响宿主时的相互作用， 免疫指导治疗干预。

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.