Human Placental CMV Infection: Global Gene Expression

描述（由申请方提供）：本申请中的研究旨在使用DNA微阵列技术全面了解胎盘中分化的细胞滋养层细胞（CTB）中的基因表达，并评估人巨细胞病毒（CMV）感染对这些细胞中表达途径的影响。我们试图通过鉴定在胎盘发育过程中CTB分化失调的病毒基因来定义CMV发病机制的新生物学。通过与DNA微阵列技术先驱Joseph DeRisi博士的实验室合作，我们建议大幅扩大母基金（AI 46657）的范围。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。这种方法将允许询问整个转录程序，而不需要先验地选择要研究的基因。因此，发现新的或意想不到的基因表达的重要变化的可能性非常高，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组织了一个基因组学和蛋白质组学的微阵列核心设施，提供微阵列打印，杂交和数据分析方面的设备，协议，软件和技术培训。在DeRisi博士实验室的协助下，我们将在含有代表14，000个人类基因和整个CMV基因组的70-mer寡核苷酸的微阵列上同时检查细胞和病毒基因表达的模式。我们的具体目标是（1）确定细胞基因表达模式在分化CTB和研究CMV感染与实验室和致病性菌株的影响和（2）确定CMV基因影响CTB表达途径。在微阵列实验中提出的表达变化将通过蛋白质和RNA水平分析以及功能测定进行独立验证，并得到父母资助。通过此次合作获得的专业知识和资源将使我们能够全面了解与CTB分化和免疫功能相关的表达途径，并加深我们对临床相关原代细胞类型中CMV发病机制的理解。微阵列技术促进的基因探索可以揭示分子诊断的新靶点，并导致阻断产前CMV感染的治疗。与CTB和致瘤细胞中的侵袭和血管生成相关的表达途径之间的相似性表明，这些研究具有额外的潜力，可用于确定可用于治疗癌症中肿瘤进展的治疗和CMV基因的新靶点。

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.