Induction of toll-like receptors and NALP/PAN/PYPAF family members by modified oligonucleotides in lung epithelial carcinoma cells.
ISIS 199044 is a chimeric 2'-O-methyl-containing oligonucleotide that produces toxicity in several cultured cell lines. Upon investigation into the mechanism of cytotoxicity, we discovered that treatment of lung epithelial carcinoma cells, A549, with ISIS 199044 and several other cytotoxic oligonucleotides induces a group of genes that are not normally expressed in these cells. These genes are involved in host response to foreign materials. Among them were toll-like receptor 7 (TLR7) and TLR9, members of the toll-like receptor family, responsible for immune response to nucleic acids and cryopyrin, a member of NALP/PAN/PYPAF family, which is known to assemble with ASC and regulate NF-kappaB activation and to modulate apoptosis. Maximal induction occurred 12-24 hours posttreatment with 500 nM oligonucleotide in the presence of Lipofectin reagent. Furthermore, we have shown that this induction is chemistry dependent; it can be negated by certain modifications, such as replacement of 2'-O-methyl with 2'-O-methoxyethyl groups or substitution of phosphorothioates with phosphodiester linkages. DNA microarray analysis identified additional genes modulated by ISIS 199044, particularly genes involved in DNA damage/repair.
Isis Pharmaceuticals, Carlsbad, CA 92008, USA.
Denis
D
Drygin
Seongjoon
S
Koo
Ranjan
R
Perera
Stephen
S
Barone
C Frank
CF
Bennett
D002277:Carcinoma / Q000188:drug therapy* / Q000378:metabolism*; D002352:Carrier Proteins / Q000096:biosynthesis*; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000378:metabolism*; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D020411:Oligonucleotide Array Sequence Analysis; D009841:Oligonucleotides / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D013329:Structure-Activity Relationship; D013997:Time Factors; D051193:Toll-Like Receptors / Q000096:biosynthesis*
D002352:Carrier Proteins; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C453881:NLRP3 protein, human; D009841:Oligonucleotides; D051193:Toll-Like Receptors