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Naegleria fowleri: differential genetic expression following treatment with Hesperidin conjugated with silver
Ruqaiyyah Siddiqui1, Kavitha Rajendran2, Bahaa Abdella2,3
1Department of Biology, Chemistry and Environmental Sciences, College of Arts and Sciences, American University of Sharjah, University City, Sharjah, United Arab Emirates.
Abstract:
Naegleria fowleri causes a deadly infection known as primary amoebic meningoencephalitis (PAM). To our knowledge, there are very few transcriptome studies conducted on these brain-eating amoebae, despite rise in the number of cases. Although the Naegleria genome has been sequenced, currently, it is not well annotated. Transcriptome level studies are needed to help understand the pathology and biology of this fatal parasitic infection. Recently, we showed that nanoparticles loaded with the flavonoid Hesperidin (HDN) are potential novel antimicrobial agents. N. fowleri trophozoites were treated with and without HDN-conjugated with silver nanoparticles (AgNPs) and silver only, and then, 50% minimum inhibitory concentration (MIC) was determined. The results revealed that the MIC of HDN-conjugated AgNPs was 12.5 microg/mL when treated for 3 h. As no reference genome exists for N. fowleri, de novo RNA transcriptome analysis using RNA-Seq and differential gene expression analysis was performed using the Trinity software. Analysis revealed that more than 2000 genes were differentially expressed in response to N. fowleri treatment with HDN-conjugated AgNPs. Some of the genes were linked to oxidative stress response, DNA repair, cell division, cell signalling and protein synthesis. The downregulated genes were linked with processes such as protein modification, synthesis of aromatic amino acids, when compared with untreated N. fowleri. Further transcriptome studies will lead to understanding of genetic mechanisms of the biology and pathogenesis and/or the identification of much needed drug candidates.
Insights
Naegleria fowleri, a brain-eating amoeba, was treated with hesperidin-conjugated silver nanoparticles. This study reveals over 2000 differentially expressed genes, offering insights into drug development for primary amoebic meningoencephalitis (PAM).
Area of Science:
- Microbiology
- Genomics
- Drug Discovery
Background:
- Naegleria fowleri causes fatal primary amoebic meningoencephalitis (PAM).
- Limited transcriptome data exists for N. fowleri, hindering understanding of its biology and pathology.
- The Naegleria genome is sequenced but poorly annotated, necessitating further transcriptomic studies.
Purpose of the Study:
- To investigate the effects of hesperidin-conjugated silver nanoparticles (HDN-AgNPs) on N. fowleri.
- To perform de novo RNA transcriptome analysis to identify differentially expressed genes.
- To explore potential therapeutic targets for PAM.
Main Methods:
- Treatment of N. fowleri trophozoites with HDN-AgNPs and silver nanoparticles.
- Determination of the 50% minimum inhibitory concentration (MIC) for HDN-AgNPs.
- De novo RNA sequencing (RNA-Seq) and differential gene expression analysis using Trinity software.
Main Results:
- The MIC of HDN-AgNPs against N. fowleri was determined to be 12.5 microg/mL after 3 hours of treatment.
- Over 2000 genes were found to be differentially expressed in response to HDN-AgNPs treatment.
- Upregulated genes were associated with oxidative stress response, DNA repair, cell division, cell signaling, and protein synthesis.
- Downregulated genes were linked to protein modification and aromatic amino acid synthesis.
Conclusions:
- HDN-AgNPs show potential as antimicrobial agents against N. fowleri.
- Transcriptome analysis provides crucial insights into the genetic mechanisms underlying N. fowleri's biology and pathogenesis.
- Further studies can identify novel drug candidates for treating PAM.
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