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.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AI053782-01
Application #
6570832
Study Section
Special Emphasis Panel (ZRG1-MBC-1 (01))
Program Officer
Beisel, Christopher E
Project Start
2002-09-30
Project End
2004-08-31
Budget Start
2002-09-30
Budget End
2003-08-31
Support Year
1
Fiscal Year
2002
Total Cost
$226,000
Indirect Cost
Name
University of California San Francisco
Department
Anatomy/Cell Biology
Type
Schools of Dentistry
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Maidji, Ekaterina; Nigro, Giovanni; Tabata, Takako et al. (2010) Antibody treatment promotes compensation for human cytomegalovirus-induced pathogenesis and a hypoxia-like condition in placentas with congenital infection. Am J Pathol 177:1298-310
Pereira, L; Maidji, E (2008) Cytomegalovirus infection in the human placenta: maternal immunity and developmentally regulated receptors on trophoblasts converge. Curr Top Microbiol Immunol 325:383-95
Tabata, Takako; Kawakatsu, Hisaaki; Maidji, Ekaterina et al. (2008) Induction of an epithelial integrin alphavbeta6 in human cytomegalovirus-infected endothelial cells leads to activation of transforming growth factor-beta1 and increased collagen production. Am J Pathol 172:1127-40
Winn, Virginia D; Haimov-Kochman, Ronit; Paquet, Agnes C et al. (2007) Gene expression profiling of the human maternal-fetal interface reveals dramatic changes between midgestation and term. Endocrinology 148:1059-79
Maidji, Ekaterina; Genbacev, Olga; Chang, Hsin-Ti et al. (2007) Developmental regulation of human cytomegalovirus receptors in cytotrophoblasts correlates with distinct replication sites in the placenta. J Virol 81:4701-12
Tabata, T; McDonagh, S; Kawakatsu, H et al. (2007) Cytotrophoblasts infected with a pathogenic human cytomegalovirus strain dysregulate cell-matrix and cell-cell adhesion molecules: a quantitative analysis. Placenta 28:527-37
Adler, Stuart P; Nigro, Giovanni; Pereira, Lenore (2007) Recent advances in the prevention and treatment of congenital cytomegalovirus infections. Semin Perinatol 31:10-8
Maidji, Ekaterina; McDonagh, Susan; Genbacev, Olga et al. (2006) Maternal antibodies enhance or prevent cytomegalovirus infection in the placenta by neonatal Fc receptor-mediated transcytosis. Am J Pathol 168:1210-26
McDonagh, Susan; Maidji, Ekaterina; Chang, Hsin-Ti et al. (2006) Patterns of human cytomegalovirus infection in term placentas: a preliminary analysis. J Clin Virol 35:210-5
Pereira, Lenore; Maidji, Ekaterina; McDonagh, Susan et al. (2005) Insights into viral transmission at the uterine-placental interface. Trends Microbiol 13:164-74

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