Inhibition of Replication and Transcription Activator and Latency-associated Nuclear Antigen of Kaposi's Sarcoma-associated Herpesvirus by Morpholino Oligomers

Overview

Journal Antiviral Res

Publisher Elsevier

Date 2006 Jul 18

PMID 16842866

Citations 32

Authors

Yan-Jin Zhang

Kai-Yu Wang

David A Stein

Deendayal Patel

Rheba Watkins

Hong M Moulton

Patrick L Iversen

David O Matson

Affiliations

Soon will be listed here.

Abstract

Kaposi's sarcoma-associated herpesvirus (KSHV) is associated with Kaposi's sarcoma and primary effusion lymphoma (PEL). The KSHV replication and transcription activator (RTA) and latency-associated nuclear antigen (LANA) play key roles in activating KSHV lytic replication and maintaining KSHV latency, respectively. Phosphorodiamidate morpholino oligomers (PMO) are similar to short single-stranded DNA oligomers, but possess a modified backbone that confers highly specific binding and resistance to nucleases. In this study, RTA and LANA mRNA in PEL cells were targeted by antisense peptide-conjugated PMO (P-PMO) in an effort to suppress KSHV replication. Highly efficient P-PMO uptake by PEL cells was observed. Treatment of PEL cells with a RTA P-PMO (RP1) reduced RTA expression in a dose-dependent and sequence-specific manner, and also caused a significant decrease in several KSHV early and late gene products, including vIL-6, vIRF-1, and ORF-K8.1A. KSHV viral DNA levels were reduced both in cells and culture supernatants of RP1 P-PMO-treated cells, indicating that KSHV lytic replication was suppressed. Treatment of BCBL-1 cells with P-PMO against LANA resulted in a reduction of LANA expression. Cell viability assays detected no cytotoxicity from P-PMO alone, within the concentration range used for the experiments in this study. These results suggest that RP1 P-PMO can specifically block KSHV replication, and further study is warranted.

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References

Lukac D, Kirshner J, Ganem D . Transcriptional activation by the product of open reading frame 50 of Kaposi's sarcoma-associated herpesvirus is required for lytic viral reactivation in B cells. J Virol. 1999; 73(11):9348-61. PMC: 112969. DOI: 10.1128/JVI.73.11.9348-9361.1999. View

Schmajuk G, Sierakowska H, Kole R . Antisense oligonucleotides with different backbones. Modification of splicing pathways and efficacy of uptake. J Biol Chem. 1999; 274(31):21783-9. DOI: 10.1074/jbc.274.31.21783. View

Ghosh C, Iversen P . Intracellular delivery strategies for antisense phosphorodiamidate morpholino oligomers. Antisense Nucleic Acid Drug Dev. 2000; 10(4):263-74. DOI: 10.1089/108729000421448. View

van den Born E, Stein D, Iversen P, Snijder E . Antiviral activity of morpholino oligomers designed to block various aspects of Equine arteritis virus amplification in cell culture. J Gen Virol. 2005; 86(Pt 11):3081-3090. DOI: 10.1099/vir.0.81158-0. View

Gao S, Boshoff C, Jayachandra S, Weiss R, Chang Y, Moore P . KSHV ORF K9 (vIRF) is an oncogene which inhibits the interferon signaling pathway. Oncogene. 1997; 15(16):1979-85. DOI: 10.1038/sj.onc.1201571. View

Ensoli B, Barillari G, Buonaguro L, Gallo R . Molecular mechanisms in the pathogenesis of AIDS-associated Kaposi's sarcoma. Adv Exp Med Biol. 1991; 303:27-38. DOI: 10.1007/978-1-4684-6000-1_4. View

Gao S, Kingsley L, Hoover D, Spira T, Rinaldo C, Saah A . Seroconversion to antibodies against Kaposi's sarcoma-associated herpesvirus-related latent nuclear antigens before the development of Kaposi's sarcoma. N Engl J Med. 1996; 335(4):233-41. DOI: 10.1056/NEJM199607253350403. View

Miller G, Heston L, Grogan E, Gradoville L, Rigsby M, Sun R . Selective switch between latency and lytic replication of Kaposi's sarcoma herpesvirus and Epstein-Barr virus in dually infected body cavity lymphoma cells. J Virol. 1997; 71(1):314-24. PMC: 191053. DOI: 10.1128/JVI.71.1.314-324.1997. View

Ballestas M, Chatis P, Kaye K . Efficient persistence of extrachromosomal KSHV DNA mediated by latency-associated nuclear antigen. Science. 1999; 284(5414):641-4. DOI: 10.1126/science.284.5414.641. View

10.

Zhu J, Trang P, Kim K, Zhou T, Deng H, Liu F . Effective inhibition of Rta expression and lytic replication of Kaposi's sarcoma-associated herpesvirus by human RNase P. Proc Natl Acad Sci U S A. 2004; 101(24):9073-8. PMC: 428475. DOI: 10.1073/pnas.0403164101. View

11.

Staskus K, Sun R, Miller G, Racz P, Jaslowski A, Metroka C . Cellular tropism and viral interleukin-6 expression distinguish human herpesvirus 8 involvement in Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman's disease. J Virol. 1999; 73(5):4181-7. PMC: 104197. DOI: 10.1128/JVI.73.5.4181-4187.1999. View

12.

Scholpp S, Brand M . Morpholino-induced knockdown of zebrafish engrailed genes eng2 and eng3 reveals redundant and unique functions in midbrain--hindbrain boundary development. Genesis. 2001; 30(3):129-33. DOI: 10.1002/gene.1047. View

13.

Kedes D, Ganem D . Sensitivity of Kaposi's sarcoma-associated herpesvirus replication to antiviral drugs. Implications for potential therapy. J Clin Invest. 1997; 99(9):2082-6. PMC: 508037. DOI: 10.1172/JCI119380. View

14.

Sergerie Y, Boivin G . Evaluation of susceptibility of human herpesvirus 8 to antiviral drugs by quantitative real-time PCR. J Clin Microbiol. 2003; 41(8):3897-900. PMC: 179795. DOI: 10.1128/JCM.41.8.3897-3900.2003. View

15.

Morcos P . Achieving efficient delivery of morpholino oligos in cultured cells. Genesis. 2001; 30(3):94-102. DOI: 10.1002/gene.1039. View

16.

Moulton H, Nelson M, Hatlevig S, Reddy M, Iversen P . Cellular uptake of antisense morpholino oligomers conjugated to arginine-rich peptides. Bioconjug Chem. 2004; 15(2):290-9. DOI: 10.1021/bc034221g. View

17.

Curreli F, Cerimele F, Muralidhar S, Rosenthal L, Cesarman E, Friedman-Kien A . Transcriptional downregulation of ORF50/Rta by methotrexate inhibits the switch of Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 from latency to lytic replication. J Virol. 2002; 76(10):5208-19. PMC: 136151. DOI: 10.1128/jvi.76.10.5208-5219.2002. View

18.

Lim C, Choi C, Choe J . Mitotic chromosome-binding activity of latency-associated nuclear antigen 1 is required for DNA replication from terminal repeat sequence of Kaposi's sarcoma-associated herpesvirus. J Virol. 2004; 78(13):7248-56. PMC: 421666. DOI: 10.1128/JVI.78.13.7248-7256.2004. View

19.

Wang S, Wu F, Fujimuro M, Zong J, Hayward S, Hayward G . Role of CCAAT/enhancer-binding protein alpha (C/EBPalpha) in activation of the Kaposi's sarcoma-associated herpesvirus (KSHV) lytic-cycle replication-associated protein (RAP) promoter in cooperation with the KSHV replication and transcription.... J Virol. 2002; 77(1):600-23. PMC: 140597. DOI: 10.1128/jvi.77.1.600-623.2003. View

20.

Friborg Jr J, Kong W, Hottiger M, Nabel G . p53 inhibition by the LANA protein of KSHV protects against cell death. Nature. 2000; 402(6764):889-94. DOI: 10.1038/47266. View