High-throughput screening for TLR3-IFN regulatory factor 3 signaling pathway modulators identifies several antipsychotic drugs as TLR inhibitors.
TLR3 is one of the major innate immune sensors of dsRNA. The signal transduction pathway activated by TLR3, upon binding to dsRNA, leads to the activation of two major transcription factors: NF-kappaB and IFN regulatory factor (IRF) 3. In an effort to identify specific chemical modulators of TLR3-IRF3 signal transduction pathway, we developed a cell-based readout system. Using the IFN-stimulated gene 56 promoter-driven firefly luciferase gene stably integrated in a TLR3-expressing HEK293 cell line, we were able to generate a cell line where treatment with dsRNA resulted in a dose-dependent induction of luciferase activity. A screen of two pharmacologically active compound libraries using this system identified a number of TLR3-IRF3 signaling pathway modulators. Among them we focused on a subset of inhibitors and characterized their mode of action. Several antipsychotic drugs, such as sertraline, trifluoperazine, and fluphenazine, were found to be direct inhibitors of the innate immune signaling pathway. These inhibitors also showed the ability to inhibit IFN-stimulated gene 56 induction mediated by TLR4 and TLR7/8 pathways. Interestingly, they did not show significant effects on TLR3-, TLR7-, and TLR8-mediated NF-kappaB activation. Detailed analysis of the signaling pathway indicated that these drugs might be exerting their inhibitory effects on IRF3 via PI3K signaling pathway. The data presented in this study provide mechanistic explanation of possible anti-inflammatory roles of some antipsychotic drugs.
Zhu
Jianzhong
J
Molecular Virology Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA.
Smith
Kevin
K
Hsieh
Paishiun N
PN
Mburu
Yvonne K
YK
Chattopadhyay
Saurabh
S
Sen
Ganes C
GC
Sarkar
Saumendra N
SN
D048868:Adaptor Proteins, Signal Transducing; D014150:Antipsychotic Agents / Q000494:pharmacology*; D002460:Cell Line; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002867:Chromones / Q000494:pharmacology; D002999:Clone Cells; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D050838:Interferon Regulatory Factor-3 / Q000037:antagonists & inhibitors* / Q000502:physiology*; D009025:Morpholines / Q000494:pharmacology; D012330:RNA, Double-Stranded / Q000037:antagonists & inhibitors / Q000494:pharmacology; D016601:RNA-Binding Proteins; D020280:Sertraline / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects* / Q000276:immunology*; D051196:Toll-Like Receptor 3 / Q000037:antagonists & inhibitors* / Q000502:physiology*; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D014268:Trifluoperazine / Q000494:pharmacology
D048868:Adaptor Proteins, Signal Transducing; D014150:Antipsychotic Agents; D002867:Chromones; C484290:IFIT1 protein, human; D050838:Interferon Regulatory Factor-3; D009025:Morpholines; D012330:RNA, Double-Stranded; D016601:RNA-Binding Proteins; C495344:TLR3 protein, human; D051196:Toll-Like Receptor 3; D014157:Transcription Factors; D014268:Trifluoperazine; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D020280:Sertraline
J Biol Chem. 2002 Aug 30;277(35):32124-32
12052830
Genes Dev. 2005 Nov 15;19(22):2668-81
16260493
J Biol Chem. 2007 Dec 21;282(51):36953-60
17932028
Nat Struct Mol Biol. 2009 Jun;16(6):652-7
19430461
Mol Immunol. 2008 May;45(10):2790-6
18339426
Nature. 2001 Oct 18;413(6857):732-8
11607032
Science. 2007 Sep 7;317(5843):1390-3
17690258
Nat Med. 2005 Apr;11(4 Suppl):S63-8
15812492
Science. 2005 Sep 16;309(5742):1857-61
16166517
Cell. 2006 Feb 24;124(4):783-801
16497588
J Exp Med. 2008 Feb 18;205(2):315-22
18227218
J Immunol. 2008 Feb 15;180(4):2054-61
18250410
Immunol Res. 2006;34(1):33-48
16720897
Pharmacol Ther. 2004 Sep;103(3):245-59
15464592
Proc Natl Acad Sci U S A. 2002 Apr 30;99(9):6322-7
11972054
Nat Immunol. 2003 May;4(5):491-6
12692549
Curr Opin Immunol. 2008 Feb;20(1):17-22
18272355
J Biomol Screen. 1999;4(2):67-73
10838414
Nat Med. 2004 Dec;10(12):1366-73
15558055
J Mol Neurosci. 2006;29(3):185-94
17085778
J Leukoc Biol. 2009 Jun;85(6):966-77
19289601
J Biol Chem. 2001 Aug 10;276(32):30178-82
11487589
J Immunol. 2008 Jan 1;180(1):483-91
18097050
Respir Res. 2007 May 03;8:35
17477857
Nature. 2006 Jan 12;439(7073):208-11
16306936
Cell. 2006 Feb 24;124(4):823-35
16497591
Immunology. 2006 Jan;117(1):11-21
16423036
Cold Spring Harb Symp Quant Biol. 1998;63:609-20
10384326
J Immunol. 2002 Dec 15;169(12):6668-72
12471095
J Virol. 2002 Jun;76(11):5532-9
11991981
Biochem J. 2008 Jan 1;409(1):275-87
17868034
Immunopharmacology. 1996 Aug;34(1):27-37
8880223
Mol Pharmacol. 2007 Oct;72(4):868-75
17615244
Eur Neuropsychopharmacol. 2007 Dec;17(12):774-80
17499975
Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11657-61
8524823
J Immunol. 2006 Nov 1;177(9):6301-7
17056560
Science. 2003 May 16;300(5622):1148-51
12702806
Nat Immunol. 2004 Jul;5(7):730-7
15208624
Cancer Res. 2004 Mar 1;64(5):1861-6
14996750
Curr Opin Pharmacol. 2008 Feb;8(1):8-13
17974478
J Immunol. 2008 Oct 15;181(8):5522-9
18832709
J Virol. 2008 Apr;82(7):3500-8
18216110
Curr Opin Pharmacol. 2003 Aug;3(4):396-403
12901949
Nature. 2006 May 4;441(7089):101-5
16625202
J Immunol. 2008 Feb 1;180(3):1818-25
18209079
Nat Immunol. 2006 Feb;7(2):131-7
16424890
Nat Biotechnol. 2006 Feb;24(2):167-75
16465162
Cytokine Growth Factor Rev. 2005 Feb;16(1):1-14
15733829
Nat Struct Mol Biol. 2004 Nov;11(11):1060-7
15502848