Progressive multifocal leukoencephalopathy (PML) is a fatal demyelinating disease caused by human neurotropic JC virus. JCV infects more than 80% of human population during the childhood and establishes a latent infection for the rest of the life. The virus reactivates under immunomodulatory conditions with an unknown mechanism, resulting in productive infection of oligodendrocytes and astrocytes in the central nervous system (CNS). JCV replicates almost exclusively in glial cells, and its promoter sequence, which has tissue- specific characteristics, tightly modulates expression of viral genome in appropriate cell types and immune conditions through communication with cellular factors. We recently identified the alternative splicing factor, SF2/ASF, as a potential regulator of JCV as its overexpression in glial cells strongly suppresses viral gene expression and replication. Unexpectedly, down-regulation of JCV by SF2/ASF is mediated at transcriptional stage, thus ascribing a novel role for SF2/ASF in the control of promoter activity. Results from our preliminary studies suggest that immune mediators secreted by PBMCs induce the expression of SF2/ASF, and inhibit the replication of JCV. These observations suggest operation of a novel immune signaling pathway between peripheral immune cells and glial cells that controls the immediate early stage of JCV gene expression during the course of viral reactivation. Our preliminary studies from JCV-infected glial cells also revealed a significant role of agnoprotein, a small viral protein crucial for JCV replication, in regulation of SF2/ASF. Expression of SF2/ASF is induced, and colocalized with agnoprotein in the cytoplasm of infected cells. Results from biochemical assays showed interaction of agnoprotein with SF2/ASF suggesting a functional cross-association between these two proteins. Results from reporter gene assays revealed that agnoprotein induces the viral transcription silenced by SF2/ASF. These observations provided us a rationale to study the role of SF2/ASF in JCV life cycle, and led us to formulate our central hypothesis which s that neuroimmune regulation of JCV replication is mediated by SF2/ASF, and viral agnoprotein interferes with this regulation. We propose to examine our hypothesis by (i) examining immune mediated regulation of SF2/ASF and replication of JCV during the latent as well as productive period of the viral infection and (ii) investigating the functional interplay between SF2/ASF and JCV agnoprotein in this regulation. We will employ molecular, cellular, and virological approaches to address these questions and examine the biological relevance of our findings by immunohistochemical evaluation of clinical samples from patients with PML disease. The information from these studies will offer a novel strategy for the treatment of PML.

Public Health Relevance

Patients undergoing immune modulatory therapies for the treatment of autoimmune diseases such as multiple sclerosis, and individuals with an impaired-immune system, most notably AIDS patients, are in the high risk group of developing progressive multifocal leukoencephalopathy (PML), a fatal demyelinating disease of the white matter caused by human neurotropic polyomavirus, JC virus. Our proposed study will increase our understanding on molecular regulation of JCV replication by immune mediators during latent as well as productive stage of the viral infection, and will determin viral and cellular players of this regulation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI101192-02
Application #
8473161
Study Section
Clinical Neuroimmunology and Brain Tumors Study Section (CNBT)
Program Officer
Park, Eun-Chung
Project Start
2012-06-01
Project End
2017-05-31
Budget Start
2013-06-01
Budget End
2014-05-31
Support Year
2
Fiscal Year
2013
Total Cost
$359,550
Indirect Cost
$124,550
Name
Temple University
Department
Neurosciences
Type
Schools of Medicine
DUNS #
057123192
City
Philadelphia
State
PA
Country
United States
Zip Code
19122
Sariyer, Ilker Kudret; Sariyer, Rahsan; Otte, Jessica et al. (2016) Pur-Alpha Induces JCV Gene Expression and Viral Replication by Suppressing SRSF1 in Glial Cells. PLoS One 11:e0156819
White, Martyn K; Sariyer, Ilker K; Gordon, Jennifer et al. (2016) Diagnostic assays for polyomavirus JC and progressive multifocal leukoencephalopathy. Rev Med Virol 26:102-14
Sariyer, Rahsan; De-Simone, Francesca Isabella; Gordon, Jennifer et al. (2016) Immune suppression of JC virus gene expression is mediated by SRSF1. J Neurovirol :
Regan, Patrick M; Sariyer, Ilker K; Langford, T Dianne et al. (2016) Morphine-induced MOR-1X and ASF/SF2 Expressions Are Independent of Transcriptional Regulation: Implications for MOR-1X Signaling. J Cell Physiol 231:1542-53
Craigie, Michael; Regan, Patrick; Otalora, Yolanda-Lopez et al. (2015) Molecular interplay between T-Antigen and splicing factor, arginine/serine-rich 1 (SRSF1) controls JC virus gene expression in glial cells. Virol J 12:196
Merabova, Nana; Sariyer, Ilker Kudret; Saribas, A Sami et al. (2015) WW domain of BAG3 is required for the induction of autophagy in glioma cells. J Cell Physiol 230:831-41
De-Simone, Francesca Isabella; Sariyer, Rahsan; Otalora, Yolanda-Lopez et al. (2015) IFN-Gamma Inhibits JC Virus Replication in Glial Cells by Suppressing T-Antigen Expression. PLoS One 10:e0129694
Saribas, A Sami; Khalili, Kamel; Sariyer, Ilker Kudret (2015) Dysregulation of autophagy by HIV-1 Nef in human astrocytes. Cell Cycle 14:2899-904
Bruno, Anna Paola; De Simone, Francesca Isabella; Iorio, Vittoria et al. (2014) HIV-1 Tat protein induces glial cell autophagy through enhancement of BAG3 protein levels. Cell Cycle 13:3640-4
Otlu, Onder; De Simone, Francesca Isabella; Otalora, Yolanda-Lopez et al. (2014) The agnoprotein of polyomavirus JC is released by infected cells: evidence for its cellular uptake by uninfected neighboring cells. Virology 468-470:88-95

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