Primary amoebic meningoencephalitis is a rare but fatal central nervous system (CNS) disease caused by the "brain-eating amoeba" Naegleria fowleri. A major obstacle is the requirement for drugs with the ability to cross the blood-brain barrier, which are used in extremely high doses, cause severe side effects, and are usually ineffective. We discovered that the 4-aminomethylphenoxy-benzoxaborole AN3057 exhibits nanomolar potency against N. fowleri, and experimental treatment of infected mice significantly prolonged survival and demonstrated a 28% relapse-free cure rate.
- Klíčová slova
- Naegleria fowleri, amoeba, antiparasitic, oxaborole, primary amoebic meningoencephalitis,
- MeSH
- amébiáza * farmakoterapie MeSH
- hematoencefalická bariéra MeSH
- meningoencefalitida * MeSH
- myši MeSH
- Naegleria fowleri * MeSH
- protozoární infekce centrálního nervového systému * farmakoterapie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: The opportunistic pathogen Naegleria fowleri establishes infection in the human brain, killing almost invariably within 2 weeks. The amoeba performs piece-meal ingestion, or trogocytosis, of brain material causing direct tissue damage and massive inflammation. The cellular basis distinguishing N. fowleri from other Naegleria species, which are all non-pathogenic, is not known. Yet, with the geographic range of N. fowleri advancing, potentially due to climate change, understanding how this pathogen invades and kills is both important and timely. RESULTS: Here, we report an -omics approach to understanding N. fowleri biology and infection at the system level. We sequenced two new strains of N. fowleri and performed a transcriptomic analysis of low- versus high-pathogenicity N. fowleri cultured in a mouse infection model. Comparative analysis provides an in-depth assessment of encoded protein complement between strains, finding high conservation. Molecular evolutionary analyses of multiple diverse cellular systems demonstrate that the N. fowleri genome encodes a similarly complete cellular repertoire to that found in free-living N. gruberi. From transcriptomics, neither stress responses nor traits conferred from lateral gene transfer are suggested as critical for pathogenicity. By contrast, cellular systems such as proteases, lysosomal machinery, and motility, together with metabolic reprogramming and novel N. fowleri proteins, are all implicated in facilitating pathogenicity within the host. Upregulation in mouse-passaged N. fowleri of genes associated with glutamate metabolism and ammonia transport suggests adaptation to available carbon sources in the central nervous system. CONCLUSIONS: In-depth analysis of Naegleria genomes and transcriptomes provides a model of cellular systems involved in opportunistic pathogenicity, uncovering new angles to understanding the biology of a rare but highly fatal pathogen.
- Klíčová slova
- Cytoskeleton, Genome sequence, Illumina, Inter-strain diversity, Lysosomal, Metabolism, Neuropathogenic, Protease, RNA-Seq,
- MeSH
- genomika MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- Naegleria fowleri * genetika MeSH
- transkriptom MeSH
- trogocytóza MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Copper is a trace metal that is necessary for all organisms but toxic when present in excess. Different mechanisms to avoid copper toxicity have been reported to date in pathogenic organisms such as Cryptococcus neoformans and Candida albicans. However, little if anything is known about pathogenic protozoans despite their importance in human and veterinary medicine. Naegleria fowleri is a free-living amoeba that occurs naturally in warm fresh water and can cause a rapid and deadly brain infection called primary amoebic meningoencephalitis (PAM). Here, we describe the mechanisms employed by N. fowleri to tolerate high copper concentrations, which include various strategies such as copper efflux mediated by a copper-translocating ATPase and upregulation of the expression of antioxidant enzymes and obscure hemerythrin-like and protoglobin-like proteins. The combination of different mechanisms efficiently protects the cell and ensures its high copper tolerance, which can be advantageous both in the natural environment and in the host. Nevertheless, we demonstrate that copper ionophores are potent antiamoebic agents; thus, copper metabolism may be considered a therapeutic target.
- Klíčová slova
- Copper, Copper-translocating ATPase, Hemerythrin, Ionophores, Naegleria fowleri, Oxidative stress,
- MeSH
- adenosintrifosfatasy metabolismus MeSH
- Amoeba MeSH
- antioxidancia fyziologie MeSH
- lidé MeSH
- měď metabolismus MeSH
- mozek MeSH
- Naegleria fowleri * fyziologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- adenosintrifosfatasy MeSH
- antioxidancia MeSH
- měď MeSH
Primary amoebic meningoencephalitis (PAM) is a rapidly fatal infection caused by the free-living amoeba Naegleria fowleri The amoeba migrates along the olfactory nerve to the brain, resulting in seizures, coma, and, eventually, death. Previous research has shown that Naegleria gruberi, a close relative of N. fowleri, prefers lipids over glucose as an energy source. Therefore, we tested several already-approved inhibitors of fatty acid oxidation alongside the currently used drugs amphotericin B and miltefosine. Our data demonstrate that etomoxir, orlistat, perhexiline, thioridazine, and valproic acid inhibited growth of N. gruberi We then tested these compounds on N. fowleri and found etomoxir, perhexiline, and thioridazine to be effective growth inhibitors. Hence, not only are lipids the preferred food source for N. gruberi, but also oxidation of fatty acids seems to be essential for growth of N. fowleri Inhibition of fatty acid oxidation could result in new treatment options, as thioridazine inhibits N. fowleri growth in concentrations that can be reached at the site of infection. It could also potentiate currently used therapy, as checkerboard assays revealed synergy between miltefosine and etomoxir. Animal testing should be performed to confirm the added value of these inhibitors. Although the development of new drugs and randomized controlled trials for this rare disease are nearly impossible, inhibition of fatty acid oxidation seems a promising strategy as we showed effectivity of several drugs that are or have been in use and that thus could be repurposed to treat PAM in the future.
- Klíčová slova
- Naegleria fowleri, Naegleria gruberi, drug targets, energy metabolism, lipid metabolism, therapy, thioridazine, treatment,
- MeSH
- amfotericin B MeSH
- mastné kyseliny MeSH
- meningoencefalitida * MeSH
- Naegleria fowleri * MeSH
- Naegleria * MeSH
- protozoární infekce centrálního nervového systému * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- amfotericin B MeSH
- mastné kyseliny MeSH
Naegleria fowleri is a single-cell organism living in warm freshwater that can become a deadly human pathogen known as a brain-eating amoeba. The condition caused by N. fowleri, primary amoebic meningoencephalitis, is usually a fatal infection of the brain with rapid and severe onset. Iron is a common element on earth and a crucial cofactor for all living organisms. However, its bioavailable form can be scarce in certain niches, where it becomes a factor that limits growth. To obtain iron, many pathogens use different machineries to exploit an iron-withholding strategy that has evolved in mammals and is important to host-parasite interactions. The present study demonstrates the importance of iron in the biology of N. fowleri and explores the plausibility of exploiting iron as a potential target for therapeutic intervention. We used different biochemical and analytical methods to explore the effect of decreased iron availability on the cellular processes of the amoeba. We show that, under iron starvation, nonessential, iron-dependent, mostly cytosolic pathways in N. fowleri are downregulated, while the metal is utilized in the mitochondria to maintain vital respiratory processes. Surprisingly, N. fowleri fails to respond to acute shortages of iron by inducing the reductive iron uptake system that seems to be the main iron-obtaining strategy of the parasite. Our findings suggest that iron restriction may be used to slow the progression of infection, which may make the difference between life and death for patients.
- MeSH
- buněčné dýchání MeSH
- fyziologická adaptace * MeSH
- mitochondrie metabolismus MeSH
- Naegleria fowleri genetika metabolismus MeSH
- regulace genové exprese * MeSH
- stopové prvky metabolismus MeSH
- železo metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- stopové prvky MeSH
- železo MeSH
This is a follow-up report on the viability of pathogenic Naegleria fowleri, Naegleria australiensis and Acanthamoeba castellanii isolates during 5 to 10 years of cryopreservation at -70 degrees C. The greatest decrease in viability occurred with N. fowleri and the least occurred with N. australiensis. At 10 years of cryostorage, viability was 21% for N. fowleri, 32% for A. castellanii and 51% for N. australiensis.
- MeSH
- Acanthamoeba castellanii * MeSH
- časové faktory MeSH
- kryoprezervace * MeSH
- Naegleria fowleri * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Primary amoebic meningoencephalitis (PAM) was induced in mice by intranasal inoculation of Naegleria fowleri (Singh et Das, 1970) to study the role of the blood vessels and lungs in the early and later stages in this disease. Upon culturing blood and lung tissue obtained at 24-, 36-, 48-, 72-, 96-, and 120-hour time periods, it was found that amoebae grew only from blood and lung tissue obtained at the 96 and 120 hour time periods. Paraffin sections of the head revealed small foci of acute inflammation and amoebae within the olfactory bulb of the central nervous system (CNS) at 24 hours. Amoebae were not observed within blood vessels of the CNS until 96 and 120 hours. Also, amoebae were observed within the connective tissue surrounding blood vessels and sutures of the skull, bone marrow, and venous sinusoids between the skull bone tables at 96 and 120 hours. No amoebae or acute inflammatory reactions were observed in the lung sections from any time period and indirect immunofluorescence microscopy was negative for N. fowleri. This study provides evidence that neither blood vessels nor lungs provide routes for N. fowleri to the CNS during the early stages of PAM and that amoebae enter veins of the CNS and bone marrow during later stages of the disease.
- MeSH
- amébiáza krev parazitologie MeSH
- bulbus olfactorius parazitologie MeSH
- cévy parazitologie MeSH
- fluorescenční protilátková technika nepřímá MeSH
- kostní dřeň parazitologie MeSH
- myši MeSH
- Naegleria fowleri růst a vývoj MeSH
- nosní sliznice parazitologie MeSH
- plíce parazitologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
This is a followup report on the viability of pathogenic Acanthamoeba castellanii, Naegleria australiensis, and N. fowleri during 5 years of cryopreservation and the virulence of N. fowleri during 30 months of cryostorage, all at -70 degrees C. The greatest decrease in viability occurred during the first year of freezing and was 10-fold greater than the average yearly decrease during years 2-5. At 5 years of cryostorage viability was 33% for A. castellanii, 38% for N. fowleri and 51% for N. australiensis. Virulence of N. fowleri did not decrease during 30 months of freezing and what appeared to be an increase in virulence during cryopreservation may be the result of reduced viability of the less virulent amebae in a culture.
- MeSH
- Acanthamoeba růst a vývoj MeSH
- amébiáza parazitologie MeSH
- kryoprezervace metody MeSH
- myši MeSH
- Naegleria fowleri růst a vývoj patogenita MeSH
- Naegleria růst a vývoj MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
It has been reported that the virulence of axenically cultivated Entamoeba histolytica increases following growth with cholesterol. The purpose of this study was to determine whether cholesterol would enhance the virulence of axenically cultivated Naegleria fowleri. Amoebae were cultivated in axenic medium with (100 micrograms/ml) or without cholesterol for 6 months and tested in mice for changes in virulence. After 6 months of continuous cultivation. N. fowleri grown with cholesterol was less virulent for mice than the same strain grown without cholesterol.
- MeSH
- cholesterol farmakologie MeSH
- myši MeSH
- Naegleria fowleri účinky léků růst a vývoj patogenita MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Názvy látek
- cholesterol MeSH
The virulence of Naegleria fowleri decreases with prolonged axenic cultivation. The decline in virulence of highly virulent, mouse-passaged LEE strain amebae was monitored during 5 years of continuous axenic cultivation. The most rapid decrease in virulence occurred during the first 2 years. Virulence could be restored to original levels by 3 serial passages in mice. The composition of axenic media also affected the virulence of N. fowleri, with a more enriched medium appearing to restore some of the virulence of a weakly virulent strain.
