title: Correcting the NLRP3 inflammasome deficiency in macrophages from autoimmune NZB mice with exon skipping antisense oligonucleotides.
abstract: Inflammasomes are molecular complexes activated by infection and cellular stress, leading to caspase-1 activation and subsequent interleukin-1β (IL-1β) processing and cell death. The autoimmune NZB mouse strain does not express NLRP3, a key inflammasome initiator mediating responses to a wide variety of stimuli including endogenous danger signals, environmental irritants and a range of bacterial, fungal and viral pathogens. We have previously identified an intronic point mutation in the Nlrp3 gene from NZB mice that generates a splice acceptor site. This leads to inclusion of a pseudoexon that introduces an early termination codon and is proposed to be the cause of NLRP3 inflammasome deficiency in NZB cells. Here we have used exon skipping antisense oligonucleotides (AONs) to prevent aberrant splicing of Nlrp3 in NZB macrophages, and this restored both NLRP3 protein expression and NLRP3 inflammasome activity. Thus, the single point mutation leading to aberrant splicing is the sole cause of NLRP3 inflammasome deficiency in NZB macrophages. The NZB mouse provides a model for addressing a splicing defect in macrophages and could be used to further investigate AON design and delivery of AONs to macrophages in vivo.
authors.affiliation: School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD, Australia.
authors.forename: Sara J
authors.initials: SJ
authors.lastname: Thygesen
authors.affiliation: School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD, Australia.
authors.forename: David P
authors.initials: DP
authors.lastname: Sester
authors.affiliation: School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD, Australia.
authors.forename: Simon O
authors.initials: SO
authors.lastname: Cridland
authors.affiliation: Western Australian Neuroscience Research Institute, Perth, WA, Australia.|Centre for Comparative Genomics, Murdoch University, Murdoch, WA, Australia.
authors.forename: Steve D
authors.initials: SD
authors.lastname: Wilton
authors.affiliation: School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD, Australia.
authors.forename: Katryn J
authors.initials: KJ
authors.lastname: Stacey
mesh_terms: D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D015551:Autoimmunity / Q000187:drug effects*; D001483:Base Sequence; D005091:Exons / Q000235:genetics*; D058847:Inflammasomes / Q000378:metabolism*; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D008810:Mice, Inbred C57BL; D008814:Mice, Inbred NZB; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein / Q000172:deficiency* / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*
chemical_list: D058847:Inflammasomes; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C492716:Nlrp3 protein, mouse; D016376:Oligonucleotides, Antisense
references.citation: Exp Cell Res. 2014 Jul 1;325(1):50-5
references.pmid: 24486759
references.citation: EMBO Mol Med. 2013 Jun;5(6):814-26
references.pmid: 23666718
references.citation: Nature. 1992 Apr 30;356(6372):768-74
references.pmid: 1574116
references.citation: Oligonucleotides. 2010 Apr;20(2):69-77
references.pmid: 20377429
references.citation: Nature. 1997 Apr 10;386(6625):619-23
references.pmid: 9121587
references.citation: Science. 2002 Aug 9;297(5583):1007-13
references.pmid: 12114529
references.citation: Eur J Immunol. 1981 Oct;11(10):805-15
references.pmid: 7308288
references.citation: Science. 2008 May 2;320(5876):674-7
references.pmid: 18403674
references.citation: Semin Arthritis Rheum. 2000 Jun;29(6):385-99
references.pmid: 10924025
references.citation: Cell Death Differ. 2013 Sep;20(9):1149-60
references.pmid: 23645208
references.citation: Mol Ther. 2009 Mar;17(3):548-53
references.pmid: 18813282
references.citation: J Immunol. 2015 Aug 1;195(3):1233-41
references.pmid: 26116505
references.citation: PLoS One. 2015 Mar 27;10(3):e0120058
references.pmid: 25816009
references.citation: Mol Ther. 2010 Jun;18(6):1218-23
references.pmid: 20332768
references.citation: BMC Evol Biol. 2012 Aug 07;12:140
references.pmid: 22871040
references.citation: Mol Ther Nucleic Acids. 2014 Sep 16;3:e193
references.pmid: 25226162
references.citation: Hum Genet. 1992 Sep-Oct;90(1-2):41-54
references.pmid: 1427786
references.citation: Mol Ther. 2007 Jan;15(1):157-66
references.pmid: 17164787
references.citation: Nature. 2006 Mar 9;440(7081):228-32
references.pmid: 16407890
references.citation: Lancet. 2011 Aug 13;378(9791):595-605
references.pmid: 21784508
references.citation: Genome Res. 2011 Oct;21(10):1563-71
references.pmid: 21750108
references.citation: RNA. 2007 Oct;13(10):1609-24
references.pmid: 17684229
references.citation: Mol Cell. 2002 Aug;10(2):417-26
references.pmid: 12191486
references.citation: Nucleic Acids Res. 1997 Feb 15;25(4):888-96
references.pmid: 9016643
references.citation: Proc Natl Acad Sci U S A. 2004 Nov 2;101(44):15700-5
references.pmid: 15505203
references.citation: J Immunol. 2006 Oct 1;177(7):4473-80
references.pmid: 16982883
references.citation: J Immunol. 2012 Apr 1;188(7):3307-14
references.pmid: 22371393
references.citation: Mol Pharmacol. 2000 Aug;58(2):380-7
references.pmid: 10908306
references.citation: Arch Biochem Biophys. 1993 Jun;303(2):474-82
references.pmid: 8390225
references.citation: Mol Biol Evol. 1999 Nov;16(11):1528-34
references.pmid: 10555284
references.citation: J Clin Invest. 2003 Aug;112(4):481-6
references.pmid: 12925686
references.citation: Hum Gene Ther. 2013 May;24(5):479-88
references.pmid: 23521559