1887

Abstract

Naegleria fowleri is a free-living amoeba found in freshwater lakes and ponds and is the causative agent of primary amoebic meningoencephalitis (PAM), a rapidly fatal disease of the central nervous system (CNS). PAM occurs when amoebae attach to the nasal epithelium and invade the CNS, a process that involves binding to, and degradation of, extracellular matrix (ECM) components. This degradation is mediated by matrix metalloproteinases (MMPs), enzymes that have been described in other pathogenic protozoa, and that have been linked to their increased motility and invasive capability. These enzymes also are upregulated in tumorigenic cells and have been implicated in metastasis of certain tumours. In the present study, in vitro experiments linked MMPs functionally to the degradation of the ECM. Gelatin zymography demonstrated enzyme activity in N. fowleri whole cell lysates, conditioned media and media collected from invasion assays. Western immunoblotting indicated the presence of the metalloproteinases MMP-2 (gelatinase A), MMP-9 (gelatinase B) and MMP-14 [membrane type-1 matrix metalloproteinase (MT1-MMP)]. Highly virulent mouse-passaged amoebae expressed higher levels of MMPs than weakly virulent axenically grown amoebae. The functional relevance of MMPs in media was indicated through the use of the MMP inhibitor, 1,10-phenanthroline. The collective in vitro results suggest that MMPs play a critical role in vivo in invasion of the CNS and that these enzymes may be amenable targets for limiting PAM.

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2017-09-28
2019-09-24
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References

  1. John DT. Primary amebic meningoencephalitis and the biology of Naegleria fowleri. Annu Rev Microbiol 1982; 36: 101– 123 [CrossRef] [PubMed]
    [Google Scholar]
  2. Marciano-Cabral F. Biology of Naegleria spp. Microbiol Rev 1988; 52: 114– 133 [PubMed]
    [Google Scholar]
  3. Jarolim KL, Mccosh JK, Howard MJ, John DT. A light microscopy study of the migration of Naegleria fowleri from the nasal submucosa to the central nervous system during the early stage of primary amebic meningoencephalitis in mice. J Parasitol 2000; 86: 50– 55 [CrossRef] [PubMed]
    [Google Scholar]
  4. Martinez AJ, Nelson EC, Jones MM, Duma RJ, Rosenblum WI. Experimental Naegleria meningoencephalitis in mice: an electron microscopy study. Lab Invest 1971; 24: 465– 475
    [Google Scholar]
  5. Jamerson M, da Rocha-Azevedo B, Cabral GA, Marciano-Cabral F. Pathogenic Naegleria fowleri and non-pathogenic Naegleria lovaniensis exhibit differential adhesion to, and invasion of, extracellular matrix proteins. Microbiology 2012; 158: 791– 803 [CrossRef] [PubMed]
    [Google Scholar]
  6. Marciano-Cabral F, Cabral GA. The immune response to Naegleria fowleri amebae and pathogenesis of infection. FEMS Immunol Med Microbiol 2007; 51: 243– 259 [CrossRef] [PubMed]
    [Google Scholar]
  7. Aldape K, Huizinga H, Bouvier J, Mckerrow J. Naegleria fowleri: characterization of a secreted histolytic cysteine protease. Exp Parasitol 1994; 78: 230– 241 [CrossRef] [PubMed]
    [Google Scholar]
  8. Mat Amin N. Proteinases in Naegleria fowleri (strain NF3), a pathogenic amoeba: a preliminary study. Trop Biomed 2004; 21: 57– 60 [PubMed]
    [Google Scholar]
  9. Bruschi F, Pinto B. The significance of matrix metalloproteinases in parasitic infections involving the central nervous system. Pathogens 2013; 2: 105– 129 [CrossRef] [PubMed]
    [Google Scholar]
  10. Matin A, Stins M, Kim KS, Khan NA. Balamuthia mandrillaris exhibits metalloprotease activities. FEMS Immunol Med Microbiol 2006; 47: 83– 91 [CrossRef] [PubMed]
    [Google Scholar]
  11. Deryugina EI, Quigley JP. Matrix metalloproteinases and tumor metastasis. Cancer Metastasis Rev 2006; 25: 9– 34 [CrossRef] [PubMed]
    [Google Scholar]
  12. Gialeli C, Theocharis AD, Karamanos NK. Roles of matrix metalloproteinases in cancer progression and their pharmacological targeting. FEBS J 2011; 278: 16– 27 [CrossRef] [PubMed]
    [Google Scholar]
  13. Mohan V, Talmi-Frank D, Arkadash V, Papo N, Sagi I. Matrix metalloproteinase protein inhibitors: highlighting a new beginning for metalloproteinases in medicine. Metalloproteinases Med 2016; 3: 31– 47 [Crossref]
    [Google Scholar]
  14. Forsyth PA, Wong H, Laing TD, Rewcastle NB, Morris DG et al. Gelatinase-A (MMP-2), gelatinase-B (MMP-9) and membrane type matrix metalloproteinase-1 (MT1-MMP) are involved in different aspects of the pathophysiology of malignant gliomas. Br J Cancer 1999; 79: 1828– 1835 [CrossRef] [PubMed]
    [Google Scholar]
  15. John A, Tuszynski G. The role of matrix metalloproteinases in tumor angiogenesis and tumor metastasis. Pathol Oncol Res 2001; 7: 14– 23 [CrossRef] [PubMed]
    [Google Scholar]
  16. Ye S. Polymorphism in matrix metalloproteinase gene promoters: implication in regulation of gene expression and susceptibility of various diseases. Matrix Biol 2000; 19: 623– 629 [CrossRef] [PubMed]
    [Google Scholar]
  17. Kleinman HK, Martin GR. Matrigel: basement membrane matrix with biological activity. Semin Cancer Biol 2005; 15: 378– 386 [CrossRef] [PubMed]
    [Google Scholar]
  18. Band RN, Balamuth W. Hemin replaces serum as a growth requirement for Naegleria. Appl Microbiol 1974; 28: 64– 65 [PubMed]
    [Google Scholar]
  19. Cline M, Marciano-Cabral F, Bradley SG. Comparison of Naegleria fowleri and Naegleria gruberi cultivated in the same nutrient medium. J Protozool 1983; 30: 387– 391 [CrossRef] [PubMed]
    [Google Scholar]
  20. John DT, Howard MJ. Virulence of Naegleria fowleri affected by axenic cultivation and passage in mice. Folia Parasitol 1993; 40: 187– 191 [PubMed]
    [Google Scholar]
  21. Vyas IK, Jamerson M, Cabral GA, Marciano-Cabral F. Identification of peptidases in highly pathogenic vs. weakly pathogenic Naegleria fowleri amebae. J Eukaryot Microbiol 2015; 62: 51– 59 [CrossRef] [PubMed]
    [Google Scholar]
  22. Pettit DA, Williamson J, Cabral GA, Marciano-Cabral F. In vitro destruction of nerve cell cultures by Acanthamoeba spp.: a transmission and scanning electron microscopy study. J Parasitol 1996; 82: 769– 777 [CrossRef] [PubMed]
    [Google Scholar]
  23. Ferrante A, Bates EJ. Elastase in the pathogenic free-living amoebae Naegleria and Acanthamoeba spp. Infect Immun 1988; 56: 3320– 3321 [PubMed]
    [Google Scholar]
  24. Piña-Vázquez C, Reyes-López M, Ortíz-Estrada G, de La Garza M, Serrano-Luna J. Host-parasite interaction: parasite-derived and -induced proteases that degrade human extracellular matrix. J Parasitol Res 2012; 2012: 1– 24 [CrossRef] [PubMed]
    [Google Scholar]
  25. Geurts N, Opdenakker G, van den Steen PE. Matrix metalloproteinases as therapeutic targets in protozoan parasitic infections. Pharmacol Ther 2012; 133: 257– 279 [CrossRef] [PubMed]
    [Google Scholar]
  26. Marciano-Cabral FM, Fulford DE. Cytopathology of pathogenic and nonpathogenic Naegleria species for cultured rat neuroblastoma cells. Appl Environ Microbiol 1986; 51: 1133– 1137 [PubMed]
    [Google Scholar]
  27. Serrano-Luna J, Cervantes-Sandoval I, Tsutsumi V, Shibayama M. A biochemical comparison of proteases from pathogenic Naegleria fowleri and non-pathogenic Naegleria gruberi. J Eukaryot Microbiol 2007; 54: 411– 417 [CrossRef] [PubMed]
    [Google Scholar]
  28. Staun-Ram E, Goldman S, Gabarin D, Shalev E. Expression and importance of matrix metalloproteinase 2 and 9 (MMP-2 and -9) in human trophoblast invasion. Reprod Biol Endocrinol 2004; 2: 59– 71 [CrossRef] [PubMed]
    [Google Scholar]
  29. Toth M, Sohail A, Fridman R. Assessment of gelatinases (MMP-2 and MMP-9) by gelatin zymography. Methods Mol Biol 2012; 878: 121– 135 [CrossRef] [PubMed]
    [Google Scholar]
  30. Dufour A, Zucker S, Sampson NS, Kuscu C, Cao J. Role of matrix metalloproteinase-9 dimers in cell migration. J Biol Chem 2010; 285: 35944– 35956 [CrossRef]
    [Google Scholar]
  31. Olson MW, Bernardo MM, Pietila M, Gervasi DC, Toth M et al. Characterization of the monomeric and dimeric forms of latent and active matrix metalloproteinase-9. J Biol Chem 2000; 275: 2661– 2668 [CrossRef]
    [Google Scholar]
  32. Qoronfleh MW, Benton B, Ignacio R, Kaboord B. Selective enrichment of membrane proteins by partition phase separation for proteomic studies. J Biomed Biotechnol 2003; 2003: 249– 255 [CrossRef] [PubMed]
    [Google Scholar]
  33. Maher PA, Singer SJ. Anomalous interaction of the acetylcholine receptor protein with the nonionic detergent Triton X-114. Proc Natl Acad Sci USA 1985; 82: 958– 962 [CrossRef] [PubMed]
    [Google Scholar]
  34. Boon L, Ugarte-Berzal E, Vandooren J, Opdenakker G. Glycosylation of matrix metalloproteases and tissue inhibitors: present state, challenges and opportunities. Biochem J 2016; 473: 1471– 1482 [CrossRef] [PubMed]
    [Google Scholar]
  35. Emmanuel M, Nakano YS, Nozaki T, Datta S. Small GTPase Rab21 mediates fibronectin induced actin reorganization in Entamoeba histolytica: implications in pathogen invasion. PLoS Pathog 2015; 11: e1004666 [CrossRef] [PubMed]
    [Google Scholar]
  36. Seano G, Primo L. Podosomes and invadopodia: tools to breach vascular basement membrane. Cell Cycle 2015; 14: 1370– 1374 [CrossRef] [PubMed]
    [Google Scholar]
  37. Lohmer LL, Kelley LC, Hagedorn EJ, Sherwood DR. Invadopodia and basement membrane invasion in vivo. Cell Adh Migr 2014; 8: 246– 255 [CrossRef] [PubMed]
    [Google Scholar]
  38. Itoh Y. MT1-MMP: a key regulator of cell migration in tissue. IUBMB Life 2006; 58: 589– 596 [CrossRef] [PubMed]
    [Google Scholar]
  39. Poincloux R, Lizárraga F, Chavrier P. Matrix invasion by tumour cells: a focus on MT1-MMP trafficking to invadopodia. J Cell Sci 2009; 122: 3015– 3024 [CrossRef] [PubMed]
    [Google Scholar]
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