HIV-associated sensory polyneuropathy and neuronal injury are associated with miRNA–455-3p induction
Eugene L. Asahchop, William G. Branton, Anand Krishnan, Patricia A. Chen, Dong Yang, Linglong Kong, Douglas W. Zochodne, Bruce J. Brew, M. John Gill, Christopher Power
Eugene L. Asahchop, William G. Branton, Anand Krishnan, Patricia A. Chen, Dong Yang, Linglong Kong, Douglas W. Zochodne, Bruce J. Brew, M. John Gill, Christopher Power
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Research Article AIDS/HIV Neuroscience

HIV-associated sensory polyneuropathy and neuronal injury are associated with miRNA–455-3p induction

Abstract

Symptomatic distal sensory polyneuropathy (sDSP) is common and debilitating in people with HIV/AIDS, leading to neuropathic pain, although the condition’s cause is unknown. To investigate biomarkers and associated pathogenic mechanisms for sDSP, we examined plasma miRNA profiles in HIV/AIDS patients with sDSP or without sDSP in 2 independent cohorts together with assessing related pathogenic effects. Several miRNAs were found to be increased in the Discovery Cohort (sDSP, n = 29; non-DSP, n = 40) by array analyses and were increased in patients with sDSP compared with patients without sDSP. miR–455-3p displayed a 12-fold median increase in the sDSP group, which was confirmed by machine learning analyses and verified by reverse transcription PCR. In the Validation Cohort (sDSP n = 16, non-DSP n = 20, healthy controls n = 15), significant upregulation of miR–455-3p was also observed in the sDSP group. Bioinformatics revealed that miR–455-3p targeted multiple host genes implicated in peripheral nerve maintenance, including nerve growth factor (NGF) and related genes. Transfection of cultured human dorsal root ganglia with miR–455-3p showed a concentration-dependent reduction in neuronal β-III tubulin expression. Human neurons transfected with miR–455-3p demonstrated reduced neurite outgrowth and NGF expression that was reversed by anti–miR–455-3p antagomir cotreatment. miR–455-3p represents a potential biomarker for HIV-associated sDSP and might also exert pathogenic effects leading to sDSP.

Authors

Eugene L. Asahchop, William G. Branton, Anand Krishnan, Patricia A. Chen, Dong Yang, Linglong Kong, Douglas W. Zochodne, Bruce J. Brew, M. John Gill, Christopher Power

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Figure 5

Immunofluorescence and in-cell immunodetection in human neurons.

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Immunofluorescence and in-cell immunodetection in human neurons. SK-N-SH...
SK-N-SH cells were differentiated for 48 hours followed by transfection with (A–C) miRNA-NC or (D–F) miR–455-3p for 48 hours. Scale bars 32 μm. After transfection, cells were fixed and immunolabeled with anti–β-III tubulin (green) or anti-NGF (red) antibodies or stained with DAPI (blue). Immunofluorescence was a single experiment in triplicates or quadruplets. For in-cell Western analyses, differentiated SK-N-SH cells were transfected with miRNA-NC, miR–455-3p, or miR–455-3p with an miR–455-3p–specific antagomir for 48 hours. After transfection, cells were fixed, incubated with (G) anti–β-III tubulin or (H) anti-NGF antibodies, and quantified by in-cell Western analysis. All in-cell Western analysis experiments were done 3 times and the results merged, averaged, and analyzed using 1-way ANOVA (Kruskal-Wallis) test. (*P < 0.05, **P < 0.01). Horizontal bars indicate the mean ± SD.