Human Placental CMV Infection: Global Gene Expression

人胎盘 CMV 感染：全局基因表达

• 批准号: 6661949
• 负责人: LENORE PALMA PEREIRA
• 金额: $ 22.73万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6661949
• 关键词: cell adhesion molecules; cell differentiation; clinical research; complementary DNA; congenital disorders; cytomegalovirus; enzyme linked immunosorbent assay; gel electrophoresis; gene expression; genetic transcription; genome; human fetus tissue; immunofluorescence technique; immunogenetics; microarray technology; northern blottings; open reading frames; placenta; spontaneous abortion; tissue /cell culture; trophoblast; virus infection mechanism; virus protein

DESCRIPTION (provided by applicant): Studies in this application are designed to use DNA microarray technology to attain a comprehensive understanding of gene expression in differentiating cytotrophoblasts (CTBs) in the placenta and to assess the impact of human cytomegalovirus (CMV) infection on expression pathways in these cells. We seek to define a new biology of CMV pathogenesis by identifying viral genes that dysregulate the differentiation of CTBs during placental development. Through a collaboration with the laboratory of Dr. Joseph DeRisi, a pioneer in DNA microarray technology, we propose to broaden the scope of the parent grant (AI46657) substantially. Our plan is to use human and CMV genomic arrays to explore gene expression pathways in CTBs before and after CMV infection. This approach will allow interrogation of the entire transcriptional program without requiring an a priori choice of which genes to study. Thus, there is a very high likelihood of discovering important changes in the expression of novel or unexpected genes, which would be very difficult without the global assessment made possible by this new technology. Dr. DeRisi has organized a microarray core facility for genomics and proteomics at UCSF that offers equipment, protocols, software, and technical training in microarray printing, hybridization, and data analysis. With assistance from the Dr. DeRisi laboratory, we will examine patterns of cellular and viral gene expression simultaneously on microarrays containing 70-mer oligonucleotides representing 14,000 human genes and the entire CMV genome. Our specific aims are (1) to determine patterns of cellular gene expression in differentiating CTBs and investigate the impact of CMV infection with laboratory and pathogenic strains and (2) to identify CMV genes that affect CTB expression pathways. Changes in expression suggested in microarray experiments will be verified independently by analysis of protein and RNA levels and functional assays with support of the parent grant. The expertise and resources available through this collaboration will enable us to achieve a global view of expression pathways associated with CTB differentiation and immune functions and to deepen our understanding of CMV pathogenesis in a clinically relevant primary cell type. Gene exploration facilitated by microarray technology could reveal novel targets for molecular diagnosis and lead to therapies that block prenatal CMV infection. Similarities between expression pathways associated with invasion and angiogenesis in CTBs and tumorigenic cells suggest the added potential of these studies to identify novel targets for therapies and CMV genes that could be used to treat tumor progression in cancer.

描述（由申请人提供）：本申请中的研究旨在使用DNA微阵列技术，以全面了解胎盘中细胞增生细胞细胞（CTB）中的基因表达，并评估人类巨细胞瘤病毒（CMV）感染对这些细胞表达途径的影响。我们试图通过鉴定胎盘发育过程中CTB分化失调的病毒基因来定义CMV发病机理的新生物学。通过与DNA微阵列技术的先驱约瑟夫·德里西（Joseph Derisi）的实验室合作，我们建议大大扩大父母赠款的范围（AI46657）。我们的计划是使用人类和CMV基因组阵列来探索CMV感染前后CTB中的基因表达途径。这种方法将允许对整个转录程序进行询问，而无需先验选择哪些基因研究。因此，发现新颖基因或意外基因表达的重要变化很有可能，如果没有这项新技术使全球评估成为可能，这将非常困难。 Derisi博士在UCSF的基因组学和蛋白质组学组织了一个微阵列核心设施，该设施提供了微阵列打印，杂交和数据分析的设备，协议，软件和技术培训。在Derisi博士实验室的帮助下，我们将在包含代表14,000个人类基因的70-Mer寡核苷酸的微阵列上同时研究细胞和病毒基因表达的模式。我们的具体目的是（1）确定分化CTB中细胞基因表达的模式，并研究CMV感染对实验室和致病菌株的影响以及（2）鉴定影响CTB表达途径的CMV基因。在微阵列实验中建议的表达变化将通过对父母赠款的支持分析蛋白质和RNA水平以及功能分析来独立验证。通过这种合作可用的专业知识和资源将使我们能够实现与CTB分化和免疫功能相关的表达途径的全球观点，并加深我们对临床相关的主要细胞类型中CMV发病机理的理解。微阵列技术促进的基因探索可以揭示分子诊断的新靶标，并导致阻断产前CMV感染的疗法。在CTB和肿瘤细胞中与侵袭和血管生成相关的表达途径之间的相似性表明，这些研究的额外潜力是鉴定可用于治疗癌症肿瘤进展的疗法和CMV基因的新靶标。