Dimethyl Fumarate Prevents HIV-Induced Lysosomal Dysfunction and Cathepsin B Release from Macrophages
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HIV-associated neurocognitive disorders (HAND) are prevalent despite combined antiretroviral therapy, affecting nearly half of HIV-infected patients worldwide. During HIV infection of macrophages secretion of the lysosomal protein, cathepsin B, is increased. Secreted cathepsin B has been shown to induce neurotoxicity. Oxidative stress is increased in HIV-infected patients, while antioxidants are decreased in monocytes from patients with HIV-associated dementia (HAD). Dimethyl fumarate (DMF), an antioxidant, has been reported to decrease HIV replication and neurotoxicity mediated by HIV-infected macrophages. Thus, we hypothesized that DMF will decrease cathepsin B release from HIV-infected macrophages by preventing oxidative stress and enhancing lysosomal function. Monocyte-derived macrophages (MDM) were isolated from healthy donors, inoculated with HIV-1ADA, and treated with DMF following virus removal. After 12 days post-infection, HIV-1 p24 and total cathepsin B levels were measured from HIV-infected MDM supernatants using ELISA; intracellular reactive oxygen and nitrogen species (ROS/RNS) were measured from MDM lysates, and functional lysosomes were assessed using a pH-dependent lysosomal dye. Neurons were incubated with serum-free conditioned media from DMF-treated MDM and neurotoxicity was determined using TUNEL assay. Results indicate that DMF reduced HIV-1 replication and cathepsin B secretion from HIV-infected macrophages in a dose-dependent manner. Also, DMF decreased intracellular ROS/RNS levels, and prevented HIV-induced lysosomal dysfunction and neuronal apoptosis. In conclusion, the improvement in lysosomal function with DMF treatment may represent the possible mechanism to reduce HIV-1 replication and cathepsin B secretion. DMF represents a potential therapeutic strategy against HAND.
KeywordsCathepsin B HIV DMF Lysosomes MDM HIV-associated neurocognitive disorders
This research was supported in part by grants from the National Institutes of Health: R25-GM061838 (LR, KC), R01MH083516 (LMM) U54MD007600 (LMM), R25-GM082406, SC1GM11369–01 (LMM), and University of Puerto Rico School of Medicine and Biomedical Sciences Deanships. We thank the Puerto Rico Clinical and Translational Research Consortium (PRCTRC) grant U54MD007587 from National Institute on Minority Health and Health Disparities (NIMHD) and the National Institute of Allergy and Infectious Diseases (NIAID) of the National Institutes of Health for the clinical support in obtaining samples from HIV-seronegative donors and for their partial support in obtaining the Nikon Eclipse E400, with a camera SPOT Insight QE and Fluorescence X-Cite Series 120 used in fluorescence assays.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
All procedure performed in studies involving human subjects were in accordance with the ethical standards the institutional review board and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all subjects included in this study. This article does not contain any studies with animals performed by any of the authors.
- Albrecht P, Bouchachia I, Goebels N, Henke N, Hofstetter HH, Issberner A, Kovacs Z, Lewerenz J, Lisak D, Maher P, Mausberg AK, Quasthoff K, Zimmermann C, Hartung HP, Methner A (2012) Effects of dimethyl fumarate on neuroprotection and immunomodulation. J Neuroinflammation 9:163. https://doi.org/10.1186/1742-2094-9-163 CrossRefPubMedPubMedCentralGoogle Scholar
- Ancuta P, Kamat A, Kunstman KJ, Kim EY, Autissier P, Wurcel A, Zaman T, Stone D, Mefford M, Morgello S, Singer EJ, Wolinsky SM, Gabuzda D (2008) Microbial translocation is associated with increased monocyte activation and dementia in AIDS patients. PLoS One 3:e2516. https://doi.org/10.1371/journal.pone.0002516 CrossRefPubMedPubMedCentralGoogle Scholar
- Anderson AM, Muñoz-Moreno JA, McClernon D, Ellis RJ, Cookson D, Clifford DB, Collier AC, Gelman BB, Marra CM, McArthur J, McCutchan J, Morgello S, Sacktor N, Simpson DM, Franklin DR, Heaton RK, Grant I, Letendre SL, CHARTER Group (2016) Prevalence and correlates of persistent HIV-1 RNA in cerebrospinal fluid during antiretroviral therapy. J Infect Dis 215:105–113. https://doi.org/10.1093/infdis/jiw505 CrossRefPubMedPubMedCentralGoogle Scholar
- Brennan MS, Matos MF, Li B, Hronowski X, Gao B, Juhasz P, Rhodes KJ, Scannevin RH (2015) Dimethyl fumarate and monoethyl fumarate exhibit differential effects on KEAP1, NRF2 activation, and glutathione depletion in vitro. PLoS One 10:e0120254. https://doi.org/10.1371/journal.pone.0120254 CrossRefPubMedPubMedCentralGoogle Scholar
- Chen X, Hui L, Geiger NH, Haughey NJ, Geiger JD (2013) Endolysosome involvement in HIV-1 transactivator protein-induced neuronal amyloid beta production. Neurobiol Aging 34:2370–2378. https://doi.org/10.1016/j.neurobiolaging.2013.04.015 CrossRefPubMedPubMedCentralGoogle Scholar
- Cross SA, Cook DR, Chi AW et al (2011) Dimethyl fumarate, an immune modulator and inducer of the antioxidant response, suppresses HIV replication and macrophage-mediated neurotoxicity: a novel candidate for HIV neuroprotection. J Immunol 187:5015–5025. https://doi.org/10.4049/jimmunol.1101868 CrossRefPubMedPubMedCentralGoogle Scholar
- Fox RJ, Miller DH, Phillips JT, Hutchinson M, Havrdova E, Kita M, Yang M, Raghupathi K, Novas M, Sweetser MT, Viglietta V, Dawson KT, CONFIRM Study Investigators (2012) Placebo-controlled phase 3 study of oral BG-12 or glatiramer in multiple sclerosis. N Engl J Med 367:1087–1097. https://doi.org/10.1056/NEJMoa1206328 CrossRefPubMedGoogle Scholar
- Gold R, Kappos L, Arnold DL, Bar-Or A, Giovannoni G, Selmaj K, Tornatore C, Sweetser MT, Yang M, Sheikh SI, Dawson KT, DEFINE Study Investigators (2012) Placebo-controlled phase 3 study of oral BG-12 for relapsing multiple sclerosis. N Engl J Med 367:1098–1107. https://doi.org/10.1056/NEJMoa1114287 CrossRefPubMedGoogle Scholar
- Kallianpur KJ, Gerschenson M, Mitchell BI, LiButti DE, Umaki TM, Ndhlovu LC, Nakamoto BK, Chow DC, Shikuma CM (2016) Oxidative mitochondrial DNA damage in peripheral blood mononuclear cells is associated with reduced volumes of hippocampus and subcortical gray matter in chronically HIV-infected patients. Mitochondrion 28:8–15. https://doi.org/10.1016/j.mito.2016.02.006 CrossRefPubMedPubMedCentralGoogle Scholar
- Kappos L, Gold R, Miller DH, MacManus DG, Havrdova E, Limmroth V, Polman CH, Schmierer K, Yousry TA, Yang M, Eraksoy M, Meluzinova E, Rektor I, Dawson KT, Sandrock AW, O'Neill GN (2008) Efficacy and safety of oral fumarate in patients with relapsing-remitting multiple sclerosis: a multicentre, randomised, double-blind, placebo-controlled phase IIb study. Lancet 372:1463–1472. https://doi.org/10.1016/S0140-6736(08)61619-0 CrossRefPubMedGoogle Scholar
- Medina DL, Fraldi A, Bouche V, Annunziata F, Mansueto G, Spampanato C, Puri C, Pignata A, Martina JA, Sardiello M, Palmieri M, Polishchuk R, Puertollano R, Ballabio A (2011) Transcriptional activation of lysosomal exocytosis promotes cellular clearance. Dev Cell 21:421–430. https://doi.org/10.1016/j.devcel.2011.07.016 CrossRefPubMedPubMedCentralGoogle Scholar
- Neuenburg JK, Brodt HR, Herndier BG et al (2002) HIV-related neuropathology, 1985 to 1999: rising prevalence of HIV encephalopathy in the era of highly active antiretroviral therapy. J Acquir Immune Defic Syndr 31:171–177. https://doi.org/10.1097/01.QAI.0000030047.72209.D3 CrossRefPubMedGoogle Scholar
- Reddy PV, Agudelo M, Atluri VS, Nair MP (2012) Inhibition of nuclear factor erythroid 2-related factor 2 exacerbates HIV-1 gp120-induced oxidative and inflammatory response: role in HIV associated neurocognitive disorder. Neurochem Res 37:1697–1706. https://doi.org/10.1007/s11064-012-0779-0 CrossRefPubMedGoogle Scholar
- Rodriguez-Franco EJ, Cantres-Rosario YM, Plaud-Valentin M, Romeu R, Rodríguez Y, Skolasky R, Meléndez V, Cadilla CL, Melendez LM (2012) Dysregulation of macrophage-secreted cathepsin B contributes to HIV-1-linked neuronal apoptosis. PLoS One 7:e36571. https://doi.org/10.1371/journal.pone.0036571 CrossRefPubMedPubMedCentralGoogle Scholar
- Saha RN, Pahan K (2007) Differential regulation of Mn-superoxide dismutase in neurons and astroglia by HIV-1 gp120: implications for HIV-associated dementia. Free Radic Biol Med 42:1866–1878. https://doi.org/10.1016/j.freeradbiomed.2007.03.022 CrossRefPubMedPubMedCentralGoogle Scholar
- Saylor D, Dickens AM, Sacktor N, Haughey N, Slusher B, Pletnikov M, Mankowski JL, Brown A, Volsky DJ, McArthur JC (2016) HIV-associated neurocognitive disorder — pathogenesis and prospects for treatment. Nat Rev Neurol 12:234–248. https://doi.org/10.1038/nrneurol.2016.27 CrossRefPubMedPubMedCentralGoogle Scholar
- Scannevin RH, Chollate S, Jung M, Shackett M, Patel H, Bista P, Zeng W, Ryan S, Yamamoto M, Lukashev M, Rhodes KJ (2012) Fumarates promote cytoprotection of central nervous system cells against oxidative stress via the nuclear factor (erythroid-derived 2)-like 2 pathway. J Pharmacol Exp Ther 341:274–284. https://doi.org/10.1124/jpet.111.190132 CrossRefPubMedGoogle Scholar
- Schifitto G, Yiannoutsos CT, Ernst T, Navia BA, Nath A, Sacktor N, Anderson C, Marra CM, Clifford DB, For the ACTG 5114 Team (2009) Selegiline and oxidative stress in HIV-associated cognitive impairment. Neurology 73:1975–1981. https://doi.org/10.1212/WNL.0b013e3181c51a48 CrossRefPubMedPubMedCentralGoogle Scholar
- Wang Q, Chuikov S, Taitano S, Wu Q, Rastogi A, Tuck S, Corey J, Lundy S, Mao-Draayer Y (2015) Dimethyl fumarate protects neural stem/progenitor cells and neurons from oxidative damage through Nrf2-ERK1/2 MAPK pathway. Int J Mol Sci 16:13885–13907. https://doi.org/10.3390/ijms160613885 CrossRefPubMedPubMedCentralGoogle Scholar
- Wilms H, Sievers J, Rickert U, Rostami-Yazdi M, Mrowietz U, Lucius R (2010) Dimethylfumarate inhibits microglial and astrocytic inflammation by suppressing the synthesis of nitric oxide, IL-1beta, TNF-alpha and IL-6 in an in-vitro model of brain inflammation. J Neuroinflammation 7:30. https://doi.org/10.1186/1742-2094-7-30 CrossRefPubMedPubMedCentralGoogle Scholar
- Xu H, Ren D (2015) Lysosomal physiology. Annu Rev Physiol 77:57–80. https://doi.org/10.1146/annurev-physiol-021014-071649 CrossRefPubMedPubMedCentralGoogle Scholar
- Zenón F, Cantres-Rosario Y, Adiga R et al (2015) HIV-infected microglia mediate cathepsin B-induced neurotoxicity. J Neurovirol. https://doi.org/10.1007/s13365-015-0358-7