Regulatory role of non-coding RNAs in herpesvirus infections
Lead Research Organisation:
University of Cambridge
Department Name: Medicine
Abstract
Herpesviruses are medically relevant causing a variety of diseases ranging from the common cold sore to cancer. During evolution these viruses have learned to usurp numerous cellular processes and counteract any host defence mechanism. Therefore, they provide ideal tools to study fundamental biological processes.
Recently, small molecules called microRNAs were identified as key regulators in multicellular organisms and some viruses, predominantly herpesviruses. Virally-encoded microRNAs may represent novel targets for highly specific antiviral drugs. Animal models are required to understand their function throughout the virus life-cycle. The human cytomegalovirus is the leading cause of birth defects resulting from congenital infections and causes severe disease in immunocompromised patients. My group established its murine model to study viral miRNA function in vivo. In close collaboration with a network of other national and international groups this project aims at elucidating the role of cytomegalovirus miRNAs and their application as targets for novel, highly specific antiviral drugs.
I developed a novel system based on metabolic labelling of nascent RNA which allows precise monitoring the kinetics of cellular and viral gene expression. In a second project I will combine this with next-generation sequencing and whole-proteome mass-spectrometry to study virus-host interaction with superior resolution.
Recently, small molecules called microRNAs were identified as key regulators in multicellular organisms and some viruses, predominantly herpesviruses. Virally-encoded microRNAs may represent novel targets for highly specific antiviral drugs. Animal models are required to understand their function throughout the virus life-cycle. The human cytomegalovirus is the leading cause of birth defects resulting from congenital infections and causes severe disease in immunocompromised patients. My group established its murine model to study viral miRNA function in vivo. In close collaboration with a network of other national and international groups this project aims at elucidating the role of cytomegalovirus miRNAs and their application as targets for novel, highly specific antiviral drugs.
I developed a novel system based on metabolic labelling of nascent RNA which allows precise monitoring the kinetics of cellular and viral gene expression. In a second project I will combine this with next-generation sequencing and whole-proteome mass-spectrometry to study virus-host interaction with superior resolution.
Technical Summary
All herpesviruses share the ability to persist for life in infected individuals thus posing the risk of reactivation and disease. The eight human herpesviruses inflict a substantial burden of disease and economic costs in immunosuppressed patients and in pregnancy. Our lack of understanding of the molecular mechanisms governing viral gene expression and manipulation of host gene expression significantly hinders our ability to deal with the associated diseases and has obstructed the development of new therapies.
Here I propose two closely related projects which ask three major questions.
1) How do herpesviruses modulate cellular and viral gene expression?
2) What is the contribution and function of herpesvirus miRNAs?
3) Can viral miRNAs serve as targets for novel antiviral drugs?
The low temporal resolution of standard gene expression profiling and the inability to precisely measure RNA decay has severely hampered our understanding of the regulation of gene expression during virus infections. We developed a novel system for nascent transcriptome analysis (NTA) to overcome all of these problems. We now combine NTA with next-generation sequencing to study regulation of gene expression during lytic MCMV and HCMV infection. Similar approaches will be applied to the ?- and ?-herpesvirus family (HSV-1, MHV68). Thereby, we will provide a comprehensive picture of the regulation of gene expression in herpesvirus infection. SILAC proteomics available at the Babraham Institute will support NTA data at whole proteome level but will also elucidate the effects of viral miRNAs on target protein synthesis.
In close collaboration with a number of international groups my group established the infection of the mouse with the murine cytomegalovirus (MCMV) to study the function of cytomegalovirus miRNAs in vivo. Technologies like traceless-mutagenesis of viral genomes, miRNA target identification by RIP-Chip as well as in vivo experiments are well established. We will now study how viral miRNAs concert with viral proteins to provide a favourable environment for these viruses in vivo and how this knowledge can be used against them.
Viral miRNA research is a highly competitive field of research. Based on the advantage of a timely start and well established collaborations with leading groups in the field I am confident that both of these studies are straightforward. The aims are realistically achievable and the outcome will greatly improve our understanding of herpesvirus biology in general. Most importantly, the methodological approaches are easily transferred to any other cellular process opening up a plethora of opportunities for the future.
Here I propose two closely related projects which ask three major questions.
1) How do herpesviruses modulate cellular and viral gene expression?
2) What is the contribution and function of herpesvirus miRNAs?
3) Can viral miRNAs serve as targets for novel antiviral drugs?
The low temporal resolution of standard gene expression profiling and the inability to precisely measure RNA decay has severely hampered our understanding of the regulation of gene expression during virus infections. We developed a novel system for nascent transcriptome analysis (NTA) to overcome all of these problems. We now combine NTA with next-generation sequencing to study regulation of gene expression during lytic MCMV and HCMV infection. Similar approaches will be applied to the ?- and ?-herpesvirus family (HSV-1, MHV68). Thereby, we will provide a comprehensive picture of the regulation of gene expression in herpesvirus infection. SILAC proteomics available at the Babraham Institute will support NTA data at whole proteome level but will also elucidate the effects of viral miRNAs on target protein synthesis.
In close collaboration with a number of international groups my group established the infection of the mouse with the murine cytomegalovirus (MCMV) to study the function of cytomegalovirus miRNAs in vivo. Technologies like traceless-mutagenesis of viral genomes, miRNA target identification by RIP-Chip as well as in vivo experiments are well established. We will now study how viral miRNAs concert with viral proteins to provide a favourable environment for these viruses in vivo and how this knowledge can be used against them.
Viral miRNA research is a highly competitive field of research. Based on the advantage of a timely start and well established collaborations with leading groups in the field I am confident that both of these studies are straightforward. The aims are realistically achievable and the outcome will greatly improve our understanding of herpesvirus biology in general. Most importantly, the methodological approaches are easily transferred to any other cellular process opening up a plethora of opportunities for the future.