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Neurotoxic effects of the HCV core protein are mediated by sustained activation of ERK via TLR2 signaling

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Abstract

Hepatitis C virus (HCV) infection is a serious problem among those co-infected with human immunodeficiency virus; however, its impact in the central nervous system (CNS) remains unclear. This study aimed to investigate the mechanisms underlying HCV core protein-mediated neurodegeneration. Analysis of human HCV seropositive cases demonstrated widespread damage to neuronal dendritic processes and sustained activation of extracellular signal-related kinase (ERK); analogous pathologies were observed in wild type injected with HCV core protein into the hippocampus. In vitro analysis in neuronal cells exposed to HCV core demonstrated retraction of the neuronal processes in an ERK/Signal Transducer and Activator of Transcription 3 (STAT3)-dependent manner dependent on toll-like receptor 2 (TLR2) signaling activation. These results indicate that HCV core protein neurotoxicity may be mediated by the sustained activation of ERK/STAT3 via TLR2-IRAK1 signaling pathway. These pathways provide novel targets for development of neuroprotective treatments for HCV involvement of the CNS.

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Acknowledgements

We would like to thank Drs. D. Smith and D. Wyles for their discussions and comments. This study is supported by NIH grants MH059745, MH62512, MH62962, and NIDA DA12065.

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Author notes

  1. Amy D. Paulino

    Present address: Neuropore Therapies Inc., 3030 Bunker Hill St. Suite 117A, San Diego, CA, 92109, USA

  2. Ian P. Everall

    Present address: Department of Psychiatry, University of Melbourne, Level 1 North, Royal Melbourne Hospital, Melbourne, VIC, 3050, Australia

Authors and Affiliations

  1. Department of Neurosciences, University of California San Diego, 9500 Gilman Drive, La Jolla, CA, 92093-0624, USA

    Amy D. Paulino, Kiren Ubhi, Edward Rockenstein, Anthony Adame, Ronald Ellis & Eliezer Masliah

  2. Department of Pathology, University of California San Diego, La Jolla, CA, USA

    Leslie Crews & Eliezer Masliah

  3. Department of Medicine, University of California San Diego, La Jolla, CA, USA

    Scott Letendre

  4. Department of Psychiatry and the HIV Neurobehavioral Research Center, University of California San Diego, La Jolla, CA, USA

    Ian P. Everall & Igor Grant

  5. Veterans Affairs Healthcare System, La Jolla, CA, USA

    Igor Grant

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Correspondence to Eliezer Masliah.

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Supplementary Fig. 1

Co-immunoprecipitation studies TLR2/IRAK pathway after HCV core exposure. a Immunoprecipitation with the IRAK1 antibody resulted in the detection of TRAF6 by immunoblot only in neuronal cells treated with HCV core compared to vehicle. b Immunoprecipitation with the TRAF6 antibody resulted in the detection of TAB1 by immunoblot (55 kDa band) in neuronal cells treated with HCV core compared to vehicle. c Immunoprecipitation with the TLR2 antibody resulted in the detection of MyD88 by immunoblot only in HCV core-challenged neuronal cells compared to vehicle controls. These effects were not observed in control experiments where the neuronal cells treated with HCV core were immunoprecipitated with a non-specific IgG (DOC 466 kb)

Supplementary Fig. 2

HCV core protein toxicity in neuronal cells derived from neuronal precursor cells. a β-tubulin and (b) pERK immunoreactivity in vehicle-treated control neuronal cells. c Co-localization of β-tubulin and pERK immunoreactivity in vehicle-treated control neuronal cells. d β-tubulin and (e) pERK immunoreactivity in HCV core-treated neuronal cells. f Co-localization of β-tubulin and pERK immunoreactivity in HCV core-treated neuronal cells. g β-tubulin and (h) pERK immunoreactivity in inactive HCV core-treated neuronal cells. i Co-localization of β-tubulin and pERK immunoreactivity in inactive HCV core-treated neuronal cells. j β-tubulin and (k) pERK immunoreactivity in HCV NS3-treated neuronal cells. l Co-localization of β-tubulin and pERK immunoreactivity in HCV NS3-treated neuronal cells. m Analysis of pERK immunoreactivity across experimental groups. Scale bar = 30 μM. Asterisk indicates p < 0.05 one-way ANOVA with post hoc Fisher. HCV hepatitis C virus, pERK phospho-extracellular signal-related kinase (DOC 1606 kb)

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Paulino, A.D., Ubhi, K., Rockenstein, E. et al. Neurotoxic effects of the HCV core protein are mediated by sustained activation of ERK via TLR2 signaling. J. Neurovirol. 17, 327–340 (2011). https://doi.org/10.1007/s13365-011-0039-0

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