Cytomegalovirus-mediated modification of host cell metabolism

巨细胞病毒介导的宿主细胞代谢修饰

• 批准号: 8084510
• 负责人: JAMES C ALWINE
• 金额: $ 32.75万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8084510
• 关键词: Address; Adipocytes; Area; Biogenesis; Cell physiology; Cells; Cellular Stress Response; Citric Acid Cycle; Cytomegalovirus; Cytomegalovirus Infections; Data; Development; Disease; Embryo; Endoplasmic Reticulum; Enzymes; Fatty Acids; Fibroblasts; Gene Activation; Genes; Glucose; Glucose Transporter; Glutamine; Herpesviridae; Human; Infection; Knock-out; Link; Lipids; Maintenance; Mammalian Cell; Mediating; Metabolism; Modification; Molecular; Molecular Virology; Murid herpesvirus 1; Mus; Nutrient; Pathogenesis; Phosphotransferases; Production; Proteins; Publishing; Research; Role; Signal Transduction; Stress Response Signaling; System; Testing; Therapeutic Intervention; Viral; Virus; Warburg Effect; Work; adipocyte differentiation; c-myc Genes; design; endoplasmic reticulum stress; glucose uptake; interest; lipid biosynthesis; neoplastic cell; overexpression; programs; research study; response; sensor; uptake; virology

DESCRIPTION (provided by applicant): HCMV infection dramatically alters glucose and glutamine metabolism. The hypothesis of this proposal is that HCMV induces adipocyte-like differentiation in infected cells in order to allow glucose to be used synthetically for the production of fatty acids and lipids which are known to be essential for HCMV replication. This requires the activation of lipogenic genes, increased glucose and glutamine uptake and the use of glutamine to maintain the tricarboxylic acid (TCA) cycle. Our data show that glutamine is essential for maintenance of the TCA cycle during HCMV infection. This is done in a manner that is very similar to many tumor cells in which glucose is used synthetically and glutamine must be used to maintain the TCA cycle. In tumor cells the program for this use of glutamine is activated by c-Myc; our preliminary data suggest that this is also the case in HCMV infected cells. Adipocyte differentiation and the activation of lipogenic genes have previously been shown to be mediated by PKR-like endoplasmic reticulum kinase (PERK), an ER-resident sensor that is activated by endoplasmic reticulum stress, specifically the unfolded protein response (UPR). Our published and preliminary studies of HCMV-mediated effects on the UPR suggest that PERK is activated by HCMV infection and induces an adipocyte-like differentiation program. The specific aims are: 1) Determine the role of PERK in the induction of lipid biosynthesis and adipocyte-like differentiation in HCMV infected human fibroblasts. 2) Determine the role of c-Myc in the induction of glutamine utilization for the maintenance of the TCA cycle in infected cells. 3) Use murine cytomegalovirus (MCMV) to further assess the roles of PERK and c-Myc in inducing lipid biogenesis and the utilization of glutamine to maintain the TCA cycle. Here we will expand the studies using MCMV with the many mouse embryo fibroblast lines with genes of interest for Aims 1 and 2 knocked out or conditionally knocked out. This approach not only provides a means to verify the studies with HCMV, but also a means to address questions that would be more difficult, or impossible, in the human system. Accomplishing the aims will shift current research toward the concept that viruses like HCMV must dramatically alter cellular metabolism and that they accomplish this by integrating mechanisms for manipulating cellular signaling and stress responses. PUBLIC HEALTH RELEVANCE: In order to better understand the molecular pathogenesis of human cytomegalovirus (HCMV) it is essential to determine the means by which the virus alters key cellular functions such as cellular metabolism. Such understanding will elucidate new aspects of HCMV virology and pathogenesis, help clarify HCMVs potential role in transformation and will highlight new targets for therapeutic intervention in HCMV disease.

描述（由申请人提供）：HCMV感染极大地改变了葡萄糖和谷氨酰胺代谢。该提议的假设是HCMV在感染细胞中诱导脂肪细胞样分化，以便允许葡萄糖合成用于产生脂肪酸和脂质，这些脂肪和脂质对于HCMV复制至关重要。这需要激活脂肪生成基因，增加的葡萄糖和谷氨酰胺摄取以及使用谷氨酰胺来维持三羧酸（TCA）周期。我们的数据表明，谷氨酰胺对于在HCMV感染过程中维持TCA周期至关重要。这是以与许多肿瘤细胞非常相似的方式完成的，在这些肿瘤细胞中使用葡萄糖，必须使用谷氨酰胺来维持TCA循环。在肿瘤细胞中，这种使用谷氨酰胺的程序被C-MYC激活。我们的初步数据表明，在HCMV感染的细胞中也是如此。脂肪细胞分化和脂肪生成基因的激活先前已被证明是由PKR样性内质网激酶（PERK）介导的，这是一种通过内质网应激激活的ER驻留传感器，该传感器是由内质网应激激活的，该传感器（特别是外向的蛋白质反应（UPR））。我们对HCMV介导的对UPR的影响的发表和初步研究表明，PERK是通过HCMV感染激活的，并诱导了类似脂肪细胞的分化程序。具体目的是：1）确定PERK在HCMV感染的人成纤维细胞中诱导脂质生物合成和脂肪细胞样分化中的作用。 2）确定C-MYC在诱导谷氨酰胺利用中维持TCA周期中受感染细胞中的作用。 3）使用鼠巨细胞病毒（MCMV）进一步评估PERK和C-MYC在诱导脂质生物发生和利用谷氨酰胺来维持TCA循环中的作用。在这里，我们将使用MCMV与许多小鼠胚胎成纤维细胞系进行扩展研究，并以目标1和2击倒或有条件地击倒目标1和2。这种方法不仅提供了一种用HCMV验证研究的方法，而且还提供了一种解决人类系统中更困难或不可能的问题的方法。实现目标将把当前的研究转移到诸如HCMV之类的病毒必须显着改变细胞代谢的概念，并通过整合操纵细胞信号和应力反应的机制来实现这一目标。 公共卫生相关性：为了更好地了解人类巨细胞病毒（HCMV）的分子发病机理，必须确定病毒改变关键细胞功能（例如细胞代谢）的手段至关重要。这种理解将阐明HCMV病毒学和发病机理的新方面，有助于阐明HCMV在转化中的潜在作用，并将重点介绍HCMV疾病治疗干预的新目标。