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Published on: June 2, 2021
Iron starvation induces apoptosis in Rhizopus oryzae in vitro
Fazal Shirazi1, Dimitrios P Kontoyiannis, Ashraf S Ibrahim
1a Department of Infectious Diseases ; Infection Control and Employee Health; The University of Texas M D Anderson Cancer Center ; Houston , TX USA.
Abstract:
Mortality associated with mucormycosis remains high despite current antifungals. Iron-starvation strategies have been shown to have promising activity against Mucorales. We hypothesized that iron starvation enhances apoptosis in Rhizopus oryzae. Apoptosis was characterized in R. oryzae transformed with RNAi plasmid targeting FTR1 expression (iron permease mutant) or empty plasmid grown in iron rich (0.125% FeCl3) and iron depleted media (YNB+1mM ferrozine and 1 mM ascorbic acid). Increased apoptosis was observed with dihydrorhodamine-123 and rhodamine-123 staining in the iron starved mutant FTR1 when compared to empty plasmid, followed by increased extracellular ATP levels. In addition, DNA fragmentation and metacaspase activity were prominent in FTR1. In contrast, Rhizopus strains grown in iron-rich medium displayed minimal apoptosis. Our results demonstrate a metacaspase dependent apoptotic process in iron deprived condition and further support the role of iron starvation strategies as an adjunct treatment for mucormycosis, a mechanism by which iron starvation affects R. oryzae.
Insights
Iron starvation induces apoptosis in Rhizopus oryzae, a key fungus in mucormycosis. This finding supports iron-deprivation as a potential adjunctive therapy to combat high mortality rates associated with this fungal infection.
Area of Science:
- Mycology
- Molecular Biology
- Infectious Diseases
Background:
- Mucormycosis poses a significant mortality risk, with limited effective antifungal treatments.
- Iron availability is crucial for fungal growth, and iron-starvation strategies show promise against Mucorales.
Purpose of the Study:
- To investigate the hypothesis that iron starvation enhances apoptosis in Rhizopus oryzae.
- To elucidate the molecular mechanisms underlying iron starvation-induced cell death in this fungus.
Main Methods:
- Utilized RNA interference (RNAi) targeting the FTR1 gene (iron permease) in R. oryzae.
- Compared apoptosis markers (dihydrorhodamine-123, rhodamine-123 staining, extracellular ATP, DNA fragmentation, metacaspase activity) in iron-starved and iron-rich conditions.
Main Results:
- Iron-starved FTR1 mutants exhibited significantly increased apoptosis compared to controls.
- Elevated extracellular ATP levels, DNA fragmentation, and metacaspase activity were observed in iron-deprived R. oryzae.
- Rhizopus strains grown in iron-rich media showed minimal signs of apoptosis.
Conclusions:
- Iron starvation triggers a metacaspase-dependent apoptotic process in R. oryzae.
- These findings support the therapeutic potential of iron starvation strategies as an adjunct treatment for mucormycosis.
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