Bernasconi
Fakultäten » Medizinische Fakultät » Kinderspital Zürich: Medizinische Klinik » Infektiologie, Abteilung » Prof. Dr. David Nadal » Bernasconi
Completed research project
| Title / Titel | Silencing of the LMP2 gene in Epstein-Barr virus transformed B cells: induction of lytic viral cycle for tumor therapy | ||||||
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| Abstract (PDF, 14 KB) | |||||||
| Summary / Zusammenfassung | This research project is guided by the hypothesis that silencing of the EBV gene LMP2 results in switching from the latent to the lytic EBV life cycle and lysis of EBV-harboring tumor cells. We will target EBV-infected tumor cells by designing an RNA interference (RNAi) system to specifically down-regulate LMP2, a protein required for EBV latency maintenance and thus tumor cell survival. RNAi, triggered by double-stranded RNA, enables gene silencing by degrading specific mRNA transcripts. Short RNAs, 19-21 nucleotides in length (siRNA), either synthetic or produced by in vitro translation, are sufficient to initiate RNAi in mammalian cells. A single dose of siRNA maintains gene silencing for up to 4 days and is sufficient to study many cell functions. We will investigate the effect of LMP2 down-regulation on tumor cell survival, proliferation, apoptosis, or necrosis. Moreover, we will use in-house developed EBV-specific real-time polymerase chain reaction-based systems and microarrays to profile and quantify EBV gene expression in cells upon LMP2 down-regulation to assess the specificity of the treatment and to identify potential new targets for down-regulation of key genes in EBV-harboring tumor cells. Affymetrix chip technology will be employed to assess effects of LMP2 gene silencing on human genes to characterize provoked host gene expression profiles which in turn may reveal key targets for alternative interventions. Since LMP2A is required for maintenance of EBV latent cycle, we hope thereby to induce or, at least, lower the threshold for lytic cycle activation. Further, should down-regulation of LMP2 prove insufficient, we foresee to develop an improved strategy to amplify the induction of lytic cycle in EBV-transformed cells. Finally, we will study the effect of LMP2 down-regulation on the infectivity of newly formed virus particles and of the impact of siRNA in preventing primary cellular infection with EBV or abrogating the establishment of EBV latency. | ||||||
| Project leadership and contacts / Projektleitung und Kontakte |
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| Funding source(s) / Unterstützt durch |
Foundation Zürcher Krebsliga |
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| Duration of Project / Projektdauer | Sep 2004 to Aug 2007 |