Impact of diverse nutrient environment on viral replication and metabolite flow in human cytomegalovirus infection

项目摘要 人巨细胞病毒(HCMV)是一种流行的疱疹病毒，可建立终身感染。巨细胞病毒 感染会导致免疫功能低下的人患上严重的疾病，是先天性心脏病的主要原因 残疾人士。与所有病毒一样，HCMV依赖宿主新陈代谢来构建病毒复制的基础，例如 用于病毒基因组合成的核苷酸、用于病毒蛋白质的氨基酸和用于病毒膜的脂类。巨细胞病毒 是高度物种特异性的，这将病毒复制的分子研究限制在细胞培养模型上 感染。高营养条件下HCMV代谢重塑寄主代谢的前期研究 重点是获得最佳的病毒生产。然而，支持最佳病毒的高营养培养基 复制不能概括人体内的营养环境。这个项目将解决这个问题 通过确定低营养环境如何改变养分利用来支持巨细胞病毒复制的缺点。我 假设人巨细胞病毒可以通过代谢在不同的营养环境中复制到次优水平 灵活性。 我是通过关注无糖状态下的葡萄糖利用和替代营养流来启动这些研究的 人巨细胞病毒感染期间的培养。初步数据表明，病毒基因组合成、病毒蛋白水平、 在缺糖的过程中，病毒的产生会减少。挽救病毒基因组水平不能恢复 病毒的产生，表明葡萄糖支持病毒复制的多个步骤。而血脂正常情况下 在HCMV复制过程中由葡萄糖制成，我发现在HCMV感染过程中脂类被合成，尽管 葡萄糖的流失，表明另一种营养流正在发生。在目标1中，我将确定葡萄糖如何 丢失会影响HCMV复制阶段。这些研究将确定病毒复制的阶段，需要 并将决定哪些营养素可以补偿葡萄糖，以支持病毒复制。在目标2中，我 将确定葡萄糖损失如何改变营养流动，以支持在HCMV复制过程中的脂肪合成。这部作品 将确定谷氨酰胺是否正在补偿葡萄糖的脂肪合成，并研究葡萄糖水平如何 影响营养物质的流动。这些研究建立在确定人巨细胞病毒代谢复杂性的先前研究的基础上 控制力。由此产生的发现将增加我们对代谢重塑和替代的理解 在复制过程中使用营养物质，同时为开发与生理更相关的模型提供洞察力 巨细胞病毒与宿主代谢的相互作用。

Project Abstract Human cytomegalovirus (HCMV) is a prevalent herpesvirus that establishes lifelong infection. HCMV infection causes severe disease in immunocompromised individuals and is a leading cause of congenital disabilities. Like all viruses, HCMV relies on host metabolism for the building blocks of viral replication, such as nucleotides for viral genome synthesis, amino acids for viral proteins, and lipids for the virus membrane. HCMV is highly species-specific, which limits molecular investigation of virus replication to cell culture models of infection. Prior research on HCMV metabolic remodeling of host metabolism utilized high nutrient culture media with a focus on obtaining optimal virus production. However, high nutrient media that supports optimal virus replication does not recapitulate the nutrient environment in the human body. This project will address this drawback by identifying how low nutrient environments alter nutrient utilization to support HCMV replication. I hypothesize that HCMV can replicate to sub-optimal levels in diverse nutrient environments via metabolic flexibility. I am initiating these studies by focusing on glucose utilization and alternative nutrient flow in glucose-free cultures during HCMV infection. Preliminary data demonstrate that viral genome synthesis, viral protein levels, and virus production are decreased during glucose deprivation. Rescuing viral genome levels does not restore virus production, suggesting that glucose supports multiple steps of virus replication. While lipids are normally made from glucose during HCMV replication, I found that lipids are synthesized during HCMV infection despite the loss of glucose, suggesting that alternative nutrient flow is occurring. In aim 1, I will determine how glucose loss impacts HCMV replication stages. These studies will define the stages of virus replication that require glucose and will determine which nutrients can compensate for glucose to support virus replication. In aim 2, I will identify how glucose loss alters nutrient flow to support lipid synthesis during HCMV replication. This work will determine if glutamine is compensating for glucose for lipid synthesis and investigate how glucose levels impact nutrient flow. These studies build upon prior research that identified the complexities of HCMV metabolic control. The resulting discoveries will increase our understanding of metabolic remodeling and alternative nutrient use during HCMV replication while providing insight for developing a more physiological-relevant model of HCMV interaction with host metabolism.