Uptake and antifungal activity of oligonucleotides in Candida albicans.
Candida albicans is a significant cause of disease in immunocompromised humans. Because the number of people infected by fungal pathogens is increasing, strategies are being developed to target RNAs in fungi. This work shows that oligonucleotides can serve as therapeutics against C. albicans. In particular, oligonucleotides are taken up from cell culture medium in an energy-dependent process. After uptake, oligonucleotides, including RNA, remain mostly intact after 12 h in culture. For culture conditions designed for mammalian cells, intracellular concentrations of oligonucleotides in C. albicans exceed those in COS-7 mammalian cells, suggesting that uptake can provide selective targeting of fungi over human cells. A 19-mer 2'OMe (oligonucleotide with a 2'-O-methyl backbone) hairpin is described that inhibits growth of a C. albicans strain at pH < 4.0. This pH is easily tolerated in some parts of the body subject to C. albicans infections. In vivo dimethyl sulfate modification of ribosomal RNA and the decreased rate of protein synthesis suggest that this hairpin's activity may be due to targeting the ribosome in a way that does not depend on base pairing. Addition of anti-C. albicans oligonucleotides to COS-7 mammalian cells has no effect on cell growth. Evidently, oligonucleotides can selectively serve as therapeutics toward C. albicans and, presumably, other pathogens. Information from genome sequencing and functional genomics studies on C. albicans and other pathogens should allow rapid design and testing of other approaches for oligonucleotide therapies.
Disney
Matthew D
MD
Department of Chemistry and Center for Human Genetics and Molecular Pediatric Disease, University of Rochester, Rochester, NY 14627-0216, USA.
Haidaris
Constantine G
CG
Turner
Douglas H
DH
D000818:Animals; D000935:Antifungal Agents / Q000378:metabolism / Q000494:pharmacology*; D001483:Base Sequence; D019556:COS Cells; D002176:Candida albicans / Q000187:drug effects* / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D005656:Fungal Proteins / Q000096:biosynthesis; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*
D000935:Antifungal Agents; D005656:Fungal Proteins; D009841:Oligonucleotides