Building on our previous work with antiviral drugs active against EBV we propose six specific aims to address fundamental issues posed by EBV infection: productive and persistent infection, latency, drug resistance, reversibility of drug effect, and cellular transformation. The first and third aims are designed to coupled investigation of new drugs or drug combinations with EBV biologic systems. Synergistic combinations (aim 2) of drugs with different modes of action may offer an approach to combat drug- resistance, reversibility and reduce cytotoxicity.
The second aim deals with the mode of action of the drugs in general and AZT in particular. Based on what we have learned from our studies with ACV, DHPG, BVdU, FMAU and FIAC we will investigate (a) drug phosphorylation, (b) role of EBV TK, (c) interaction of EBV polymerase with drug-triphosphates, (d) chain termination, (e) drug incorporation, (f) reversibility, (g) role of thymidylate kinase, and (h) drug-binding domains. In the fourth aim we will use mutated EBV polymerase-expressing cell lines generated by site- directed mutagenesis to study cross-resistance to the new drugs. In the fifth aim we present a plan to disrupt or cure EBV episomal maintenance with synthetic EBNA-1 antisense methylated oligonucleotides by blocking synthesis of EBNA-1 protein needed for binding to the EBV ori-P.
The final aim deals with blocking EBV-induced transformation by selected new drugs and interferon alpha and gamma, coupled with use of EBNA-1 antisense oligonucleotides to reduce episomal numbers during transformation. Data obtained from these studies will facilitate understanding of mechanisms of drug action, help in the development of selective antiviral agents, and open new approaches to dealing with EBV infection.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI017205-12
Application #
3127028
Study Section
Virology Study Section (VR)
Project Start
1980-08-01
Project End
1993-07-31
Budget Start
1991-08-01
Budget End
1992-07-31
Support Year
12
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Type
Schools of Medicine
DUNS #
078861598
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Pagano, J S; Jimenez, G; Sung, N S et al. (1992) Epstein-Barr viral latency and cell immortalization as targets for antisense oligomers. Ann N Y Acad Sci 660:107-16
Lin, J C; Reefschlager, J; Herrmann, G et al. (1992) Structure-activity relationship between (E)-5-(2-bromovinyl)- and 5-vinyl-1-beta-D-arabinofuranosyluracil (BV-araU, V-araU) in inhibition of Epstein-Barr virus replication. Antiviral Res 17:43-52
Lin, J C; De Clercq, E; Pagano, J S (1991) Inhibitory effects of acyclic nucleoside phosphonate analogs, including (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine, on Epstein-Barr virus replication. Antimicrob Agents Chemother 35:2440-3
Lin, J C; Sista, N D; Besencon, F et al. (1991) Identification and functional characterization of Epstein-Barr virus DNA polymerase by in vitro transcription-translation of a cloned gene. J Virol 65:2728-31
Lin, J C; Zhang, Z X; Chou, T C et al. (1989) Synergistic inhibition of Epstein-Barr virus: transformation of B lymphocytes by alpha and gamma interferon and by 3'-azido-3'-deoxythymidine. J Infect Dis 159:248-54
Lin, J C; Machida, H (1988) Comparison of two bromovinyl nucleoside analogs, 1-beta-D-arabinofuranosyl-E-5-(2-bromovinyl)uracil and E-5-(2-bromovinyl)-2'-deoxyuridine, with acyclovir in inhibition of Epstein-Barr virus replication. Antimicrob Agents Chemother 32:1068-72
Lin, J C; Zhang, Z X; Smith, M C et al. (1988) Anti-human immunodeficiency virus agent 3'-azido-3'-deoxythymidine inhibits replication of Epstein-Barr virus. Antimicrob Agents Chemother 32:265-7
Rompalo, A M; Roberts, P; Johnson, K et al. (1988) Empirical therapy for the management of acute proctitis in homosexual men. JAMA 260:348-53
Lin, J C; Raab-Traub, N (1987) Two strains of Epstein-Barr virus (B95-8 and a P3HR-1 subclone) that lack defective genomes induce early antigen and cause abortive infection of Raji cells. J Virol 61:1985-91
Lin, J C; DeClercq, E; Pagano, J S (1987) Novel acyclic adenosine analogs inhibit Epstein-Barr virus replication. Antimicrob Agents Chemother 31:1431-3

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