Molecular Medicine

, Volume 23, Issue 1, pp 24–33 | Cite as

miR-155 Modifies Inflammation, Endothelial Activation and Blood-Brain Barrier Dysfunction in Cerebral Malaria

  • Kevin R. Barker
  • Ziyue Lu
  • Hani Kim
  • Ying Zheng
  • Junmei Chen
  • Andrea L. Conroy
  • Michael Hawkes
  • Henry S. Cheng
  • Makon-Sébastien Njock
  • Jason E. Fish
  • John M. Harlan
  • Jose A. López
  • W. Conrad Liles
  • Kevin C. Kain
Research Article


miR-155 has been shown to participate in host response to infection and neuroinflammation via negative regulation of blood-brain barrier (BBB) integrity and T cell function. We hypothesized that miR-155 may contribute to the pathogenesis of cerebral malaria (CM). To test this hypothesis, we used a genetic approach to modulate miR-155 expression in an experimental model of cerebral malaria (ECM). In addition, an engineered endothelialized microvessel system and serum samples from Ugandan children with CM were used to examine anti-miR-155 as a potential adjunctive therapeutic for severe malaria. Despite higher parasitemia, survival was significantly improved in miR-155−/− mice versus wild-type littermate mice in ECM. Improved survival was associated with preservation of BBB integrity and reduced endothelial activation, despite increased levels of proinflammatory cytokines. Pretreatment with antagomir-155 reduced vascular leak induced by human CM sera in an ex vivo endothelial microvessel model. These data provide evidence supporting a mechanistic role for miR-155 in host response to malaria via regulation of endothelial activation, microvascular leak and BBB dysfunction in CM.



This study was supported in part by the Canadian Institutes of Health Research (CIHR MOP-13721, MOP-136813 and MOP-115160 and a CIHR Foundation grant to KCK); Canadian Vascular Network Seed Funding and CIHR (MOP-119506 to JEF); Canadian Research Chairs (to KCK, WCL and JEF); and kind donations from the Tesari Foundation and Kim Kertland. Studentship provided by Peterborough KM Hunter Charitable Foundation Graduate Awards and the McCuaig-Throop Bursary (to KRB) and Canadian Vascular Network Scholarships (to HSC and MSN). We thank Dr. Lena Serghides for technical expertise.

Supplementary material

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  1. 1.
    GBD 2015 Disease and Injury Incidence and Prevalence Collaborators. (2016) Global, regional, and national incidence, prevalence, and years lived with disability for 310 diseases and injuries, 1990–2015: a systematic analysis for the Global Burden of Disease Study 2015. Lancet. 388:1545–1602.CrossRefGoogle Scholar
  2. 2.
    Wang H, et al. (2016) Global, regional, and national life expectancy, all-cause mortality, and cause-specific mortality for 249 causes of death, 1980–2015: a systematic analysis for the Global Burden of Disease Study 2015. Lancet. 388:1459–1544.CrossRefGoogle Scholar
  3. 3.
    Dondorp AM, et al. (2010) Artesunate versus quinine in the treatment of severe falciparum malaria in African children (AQUAMAT): an open-label, randomised trial. Lancet. 376:1647–57.CrossRefGoogle Scholar
  4. 4.
    Dondorp A, Nosten F, Stepniewska K, Day N, White N. (2010) Artesunate versus quinine for treatment of severe falciparum malaria: a randomised trial. Lancet. 366:717–25.Google Scholar
  5. 5.
    Kim H, Higgins S, Liles WC, Kain KC. (2011) Endothelial activation and dysregulation in malaria: a potential target for novel therapeutics. Curr. Opin. Hematol. 18:177–85.CrossRefGoogle Scholar
  6. 6.
    Miller LH, Ackerman HC, Su X, Wellems TE. (2013) Malaria biology and disease pathogenesis: insights for new treatments. Nat. Med. 19:156–67.CrossRefGoogle Scholar
  7. 7.
    Lee WL, Liles WC (2011) Endothelial activation, dysfunction and permeability during severe infections. Curr. Opin. Hematol. 18:191–6.CrossRefGoogle Scholar
  8. 8.
    Page A V, Liles WC (2013) Biomarkers of endothelial activation/dysfunction in infectious diseases. Virulence. 4:507–16.CrossRefGoogle Scholar
  9. 9.
    Serghides L, et al. (2011) Inhaled nitric oxide reduces endothelial activation and parasite accumulation in the brain, and enhances survival in experimental cerebral malaria. PLoS One. 6:e27714.CrossRefGoogle Scholar
  10. 10.
    Yeo TW, et al. (2008) Angiopoietin-2 is associated with decreased endothelial nitric oxide and poor clinical outcome in severe falciparum malaria. Proc. Natl. Acad. Sci. USA. 105:17097–102.CrossRefGoogle Scholar
  11. 11.
    Erdman LK, et al. (2011) Combinations of host biomarkers predict mortality among Ugandan children with severe malaria: a retrospective case-control study. PLoS One. 6:e17440.CrossRefGoogle Scholar
  12. 12.
    Hughes DP, Marron MB, Brindle NPJ. (2003) The antiinflammatory endothelial tyrosine kinase Tie2 interacts with a novel nuclear factor-kappaB inhibitor ABIN-2. Circ. Res. 92:630–36.CrossRefGoogle Scholar
  13. 13.
    Kim I, et al. (2001) Vascular endothelial growth factor expression of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and E-selectin through nuclear factor-kappa B activation in endothelial cells. J. Biol. Chem. 276:7614–20.CrossRefGoogle Scholar
  14. 14.
    Gavard J, Patel V, Gutkind JS. (2008) Angiopoietin-1 prevents VEGF-induced endothelial permeability by sequestering Src through mDia. Dev. Cell. 14:25–36.CrossRefGoogle Scholar
  15. 15.
    Kontos CD, Cha EH, York JD, Peters KG. (2002) The endothelial receptor tyrosine kinase Tie1 activates phosphatidylinositol 3-kinase and Akt to inhibit apoptosis. Mol. Cell. Biol. 22: 1704–13.CrossRefGoogle Scholar
  16. 16.
    Fiedler U, et al. (2006) Angiopoietin-2 sensitizes endothelial cells to TNF-alpha and has a crucial role in the induction of inflammation. Nat. Med. 12:235–9.CrossRefGoogle Scholar
  17. 17.
    Parikh SM, et al. (2006) Excess circulating angiopoietin-2 may contribute to pulmonary vascular leak in sepsis in humans. PLoS Med. 3:e46.CrossRefGoogle Scholar
  18. 18.
    Engels BM, Hutvagner G. (2006) Principles and effects of microRNA-mediated post-transcriptional gene regulation. Oncogene. 25:6163–9.CrossRefGoogle Scholar
  19. 19.
    Cannella D, et al. (2014) miR-146a and miR-155 delineate a MicroRNA fingerprint associated with Toxoplasma persistence in the host brain. Cell Rep. 6:928–37.CrossRefGoogle Scholar
  20. 20.
    Lopez-Ramirez MA, et al. (2014) MicroRNA-155 negatively affects blood-brain barrier function during neuroinflammation. FASEB J. 28:2551–65.CrossRefGoogle Scholar
  21. 21.
    Rodriguez A, et al. (2007) Requirement of bic/microRNA-155 for normal immune function. Science. 316:608–11.CrossRefGoogle Scholar
  22. 22.
    Finney CA, et al. (2011) S1P is associated with protection in human and experimental cerebral malaria. Mol. Med. 17:717–25.CrossRefGoogle Scholar
  23. 23.
    Njock M–S, et al. (2015) Endothelial cells suppress monocyte activation through secretion of extracellular vesicles containing anti-inflammatory microRNAs. Blood. 125:3202–12.CrossRefGoogle Scholar
  24. 24.
    Vandesompele J, et al. (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 3:RESEARCH0034.CrossRefGoogle Scholar
  25. 25.
    Cheng HS, et al. (2013) MicroRNA-146 represses endothelial activation by inhibiting pro-inflammatory pathways. EMBO Mol. Med. 5:949–66.CrossRefGoogle Scholar
  26. 26.
    Hawkes MT, et al. (2015) Inhaled nitric oxide as adjunctive therapy for severe malaria: a randomized controlled trial. Malar. J. 14:421.CrossRefGoogle Scholar
  27. 27.
    Zheng Y, et al. (2012) In vitro microvessels for the study of angiogenesis and thrombosis. Proc. Natl. Acad. Sci. USA. 109:9342–7.CrossRefGoogle Scholar
  28. 28.
    Min-Oo G, et al. (2003) Pyruvate kinase deficiency in mice protects against malaria. Nat. Genet. 35:357–62.CrossRefGoogle Scholar
  29. 29.
    Ayi K, et al. (2008) Pyruvate kinase deficiency and malaria. N. Engl. J. Med. 358:1805–10.CrossRefGoogle Scholar
  30. 30.
    Patel SN, et al. (2008) C5 deficiency and C5a or C5aR blockade protects against cerebral malaria. J. Exp. Med. 205:1133–43.CrossRefGoogle Scholar
  31. 31.
    Turner GD, et al. (1994) An immunohistochemical study of the pathology of fatal malaria. Evidence for widespread endothelial activation and a potential role for intercellular adhesion molecule-1 in cerebral sequestration. Am. J. Pathol. 145:1057–69.PubMedPubMedCentralGoogle Scholar
  32. 32.
    Conroy AL, et al. (2010) Endothelium-based biomarkers are associated with cerebral malaria in Malawian children: a retrospective case-control study. PLoS One. 5:e15291.CrossRefGoogle Scholar
  33. 33.
    Grau GE, et al. (1989) Tumor necrosis factor and disease severity in children with falciparum malaria. N. Engl. J. Med. 320:1586–91.CrossRefGoogle Scholar
  34. 34.
    Krützfeldt J, et al. (2005) Silencing of microRNAs in vivo with “antagomirs.” Nature. 438:685–9.CrossRefGoogle Scholar
  35. 35.
    Brown H, et al. (2001) Blood-brain barrier function in cerebral malaria in Malawian children. Am. J. Trop. Med. Hyg. 64:207–13.CrossRefGoogle Scholar
  36. 36.
    Seydel KB, et al. (2015) Brain swelling and death in children with cerebral malaria. N. Engl. J. Med. 372:1126–37.CrossRefGoogle Scholar
  37. 37.
    Broermann A, et al. (2011) Dissociation of VE-PTP from VE-cadherin is required for leukocyte extravasation and for VEGF-induced vascular permeability in vivo. J. Exp. Med. 208:2393–401.CrossRefGoogle Scholar
  38. 38.
    Creemers EE, Tijsen AJ, Pinto YM. (2012) Circulating MicroRNAs: Novel Biomarkers and Extracellular Communicators in Cardiovascular Disease? Circ. Res. 110:483–95.CrossRefGoogle Scholar
  39. 39.
    Singh R, Pochampally R, Watabe K, Lu Z, Mo Y-Y. (2014) Exosome-mediated transfer of miR-10b promotes cell invasion in breast cancer. Mol. Cancer. 13:256.CrossRefGoogle Scholar
  40. 40.
    Alexander M, et al. (2015) Exosome-delivered microRNAs modulate the inflammatory response to endotoxin. Nat. Commun. 6:7321.CrossRefGoogle Scholar
  41. 41.
    Higgins SJ, Silver KL, Liles WC, Kain KC. (2013) Investigating the Angiopoeitin-tie2 Pathway as a Therapeutic Target to Improve Survival Following Experimental Life-Threatening Plasmodium Challenge [Abstract 1015]. In: The American Journal of Tropical Medicine and Hygiene 62nd Annual Meeting Abstract Book. Washington, DC. 89:308.Google Scholar
  42. 42.
    Lovegrove FE, et al. (2009) Serum angiopoietin-1 and -2 levels discriminate cerebral malaria from uncomplicated malaria and predict clinical outcome in African children. PLoS One. 4:e4912.CrossRefGoogle Scholar
  43. 43.
    Pamplona A, et al. (2007) Heme oxygenase-1 and carbon monoxide suppress the pathogenesis of experimental cerebral malaria. Nat. Med. 13:703–10.CrossRefGoogle Scholar
  44. 44.
    Walther M, et al. (2012) HMOX1 gene promoter alleles and high HO-1 levels are associated with severe malaria in Gambian children. PLoS Pathog. 8:e1002579.CrossRefGoogle Scholar
  45. 45.
    Serghides L, et al. (2014) PPARγ agonists improve survival and neurocognitive outcomes in experimental cerebral malaria and induce neuroprotective pathways in human malaria. PLoS Pathog. 10:e1003980.CrossRefGoogle Scholar
  46. 46.
    Linares M, et al. (2013) Glutathione peroxidase contributes with heme oxygenase-1 to redox balance in mouse brain during the course of cerebral malaria. Biochim. Biophys. Acta. 1832: 2009–18.CrossRefGoogle Scholar
  47. 47.
    Kitamuro T, et al. (2003) Bach1 Functions as a Hypoxia-inducible Repressor for the Heme Oxygenase-1 Gene in Human Cells. J. Biol. Chem. 278:9125–33.CrossRefGoogle Scholar
  48. 48.
    Davudian S, Mansoori B, Shajari N, Mohammadi A, Baradaran B. (2016) BACH1, the master regulator gene: A novel candidate target for cancer therapy. Gene. 588:30–7.CrossRefGoogle Scholar
  49. 49.
    Pulkkinen KH, Ylä-Herttuala S, Levonen A-L. (2011) Heme oxygenase 1 is induced by miR-155 via reduced BACH1 translation in endothelial cells. Free Radic. Biol. Med. 51:2124–31.CrossRefGoogle Scholar
  50. 50.
    Cheng HS, Njock M-S, Khyzha N, Dang LT, Fish JE. (2014) Noncoding RNAs regulate NF-κB signaling to modulate blood vessel inflammation. Front. Genet. 5:422.CrossRefGoogle Scholar
  51. 51.
    Baumjohann D, Ansel KM. (2013) MicroRNA-mediated regulation of T helper cell differentiation and plasticity. Nat. Rev. Immunol. 13:666–78.CrossRefGoogle Scholar
  52. 52.
    Gracias DT, et al. (2013) The microRNA miR-155 controls CD8(+) T cell responses by regulating interferon signaling. Nat. Immunol. 14:593–602.CrossRefGoogle Scholar
  53. 53.
    Seddiki N, Brezar V, Ruffin N, Lévy Y, Swaminathan S. (2014) Role of miR-155 in the regulation of lymphocyte immune function and disease. Immunology. 142:32–8.CrossRefGoogle Scholar
  54. 54.
    Howland SW, Claser C, Poh CM, Gun SY, Rénia L. (2015) Pathogenic CD8+ T cells in experimental cerebral malaria. Semin. Immunopathol. 37:221–31.CrossRefGoogle Scholar
  55. 55.
    Poh CM, Howland SW, Grotenbreg GM, Rénia L. (2014) Damage to the blood-brain barrier during experimental cerebral malaria results from synergistic effects of CD8+ T cells with different specificities. Infect. Immun. 82:4854–64.CrossRefGoogle Scholar
  56. 56.
    Guermonprez P, et al. (2013) Inflammatory Flt3l is essential to mobilize dendritic cells and for T cell responses during Plasmodium infection. Nat. Med. 19:730–8.CrossRefGoogle Scholar
  57. 57.
    Gordon EB, et al. (2015) Inhibiting the mammalian target of rapamycin blocks the development of experimental cerebral malaria. MBio. 6:e00725.CrossRefGoogle Scholar
  58. 58.
    Conroy AL, et al. (2012) Angiopoietin-2 levels are associated with retinopathy and predict mortality in Malawian children with cerebral malaria. Crit. Care Med. 40:952–9.CrossRefGoogle Scholar

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Authors and Affiliations

  • Kevin R. Barker
    • 1
    • 2
  • Ziyue Lu
    • 2
  • Hani Kim
    • 2
  • Ying Zheng
    • 3
  • Junmei Chen
    • 4
  • Andrea L. Conroy
    • 2
  • Michael Hawkes
    • 5
  • Henry S. Cheng
    • 1
    • 6
  • Makon-Sébastien Njock
    • 6
  • Jason E. Fish
    • 1
    • 6
  • John M. Harlan
    • 7
  • Jose A. López
    • 4
    • 7
  • W. Conrad Liles
    • 7
  • Kevin C. Kain
    • 1
    • 2
  1. 1.Department of Laboratory Medicine and PathobiologyUniversity of TorontoTorontoCanada
  2. 2.SAR Laboratories, Sandra Rotman Centre for Global Health, MaRS Centre, University Health Network-Toronto General Hospital and Tropical Disease Unit, Department of MedicineUniversity of TorontoTorontoCanada
  3. 3.Department of Bioengineering and Center of Cardiovascular Biology, Institute of Stem Cell and Regenerative MedicineUniversity of WashingtonSeattleUSA
  4. 4.Bloodworks Northwest Research InstituteSeattleUSA
  5. 5.Division of Infectious Diseases, Department of PediatricsUniversity of AlbertaEdmontonCanada
  6. 6.Toronto General Research InstituteUniversity Health Network, and Heart and Stroke Richard Lewar Centre of Excellence in Cardiovascular ResearchTorontoCanada
  7. 7.Department of MedicineUniversity of WashingtonSeattleUSA

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