Ameliorating effects of Gö6976, a pharmacological agent that inhibits protein kinase D, on collagen-induced arthritis

Autoři: Tae Won Yoon aff001;  Young-In Kim aff002;  Hongsik Cho aff003;  David D. Brand aff004;  Edward F. Rosloniec aff004;  Linda K. Myers aff002;  Arnold E. Postlethwaite aff004;  Karen A. Hasty aff003;  John M. Stuart aff004;  Ae-Kyung Yi aff001
Působiště autorů: Department of Microbiology, Immunology and Biochemistry, University of Tennessee Health Science Center, Memphis, Tennessee, United States of America aff001;  Department of Pediatrics, University of Tennessee Health Science Center, Memphis, Tennessee, United States of America aff002;  Department of Orthopedic Surgery and Biomedical Engineering, University of Tennessee Health Science Center, Memphis, Tennessee, United States of America aff003;  Department of Medicine, University of Tennessee Health Science Center, Memphis, Tennessee, United States of America aff004;  Veterans Affairs Medical Center-Memphis, Memphis, Tennessee, United States of America aff005
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article


Toll-like receptor (TLR) signaling can contribute to the pathogenesis of arthritis. Disruption of TLR signaling at early stages of arthritis might thereby provide an opportunity to halt the disease progression and ameliorate outcomes. We previously found that Gö6976 inhibits TLR-mediated cytokine production in human and mouse macrophages by inhibiting TLR-dependent activation of protein kinase D1 (PKD1), and that PKD1 is essential for proinflammatory responses mediated by MyD88-dependent TLRs. In this study, we investigated whether PKD1 contributes to TLR-mediated proinflammatory responses in human synovial cells, and whether Gö6976 treatment can suppress the development and progression of type II collagen (CII)-induced arthritis (CIA) in mouse. We found that TLR/IL-1R ligands induced activation of PKD1 in human fibroblast-like synoviocytes (HFLS). TLR/IL-1R-induced expression of cytokines/chemokines was substantially inhibited in Gö6976-treated HFLS and PKD1-knockdown HFLS. In addition, serum levels of anti-CII IgG antibodies, and the incidence and severity of arthritis after CII immunization were significantly reduced in mice treated daily with Gö6976. Synergistic effects of T-cell receptor and TLR, as well as TLR alone, on spleen cell proliferation and cytokine production were significantly inhibited in the presence of Gö6976. Our results suggest a possibility that ameliorating effects of Gö6976 on CIA may be due to its ability to inhibit TLR/IL-1R-activated PKD1, which might play an important role in proinflammatory responses in arthritis, and that PKD1 could be a therapeutic target for inflammatory arthritis.

Klíčová slova:

Antibodies – Arthritis – Cytokines – Enzyme-linked immunoassays – Rheumatoid arthritis – Skeletal joints – Spleen – Toll-like receptors


1. Firestein GS. Evolving concepts of rheumatoid arthritis. Nature. 2003;423(6937):356–61. doi: 10.1038/nature01661 12748655.

2. van der Heijden IM, Wilbrink B, Tchetverikov I, Schrijver IA, Schouls LM, Hazenberg MP, et al. Presence of bacterial DNA and bacterial peptidoglycans in joints of patients with rheumatoid arthritis and other arthritides. Arthritis Rheum. 2000;43(3):593–8. doi: 10.1002/1529-0131(200003)43:3<593::AID-ANR16>3.0.CO;2-1 10728753.

3. Takeda K, Kaisho T, Akira S. Toll-like receptors. Annu Rev Immunol. 2003;21:335–76. doi: 10.1146/annurev.immunol.21.120601.141126 12524386.

4. Johnson GB, Brunn GJ, Kodaira Y, Platt JL. Receptor-mediated monitoring of tissue well-being via detection of soluble heparan sulfate by Toll-like receptor 4. J Immunol. 2002;168(10):5233–9. doi: 10.4049/jimmunol.168.10.5233 11994480.

5. Midwood K, Sacre S, Piccinini AM, Inglis J, Trebaul A, Chan E, et al. Tenascin-C is an endogenous activator of Toll-like receptor 4 that is essential for maintaining inflammation in arthritic joint disease. Nat Med. 2009;15(7):774–80. Epub 2009/06/30. nm.1987 [pii] doi: 10.1038/nm.1987 19561617.

6. Okamura Y, Watari M, Jerud ES, Young DW, Ishizaka ST, Rose J, et al. The extra domain A of fibronectin activates Toll-like receptor 4. J Biol Chem. 2001;276(13):10229–33. doi: 10.1074/jbc.M100099200 11150311.

7. Abdollahi-Roodsaz S, Joosten LA, Koenders MI, Devesa I, Roelofs MF, Radstake TR, et al. Stimulation of TLR2 and TLR4 differentially skews the balance of T cells in a mouse model of arthritis. J Clin Invest. 2008;118(1):205–16. doi: 10.1172/JCI32639 18060042.

8. Deng GM, Nilsson IM, Verdrengh M, Collins LV, Tarkowski A. Intra-articularly localized bacterial DNA containing CpG motifs induces arthritis. Nat Med. 1999;5(6):702–5. doi: 10.1038/9554 10371511.

9. Joosten LA, Koenders MI, Smeets RL, Heuvelmans-Jacobs M, Helsen MM, Takeda K, et al. Toll-like receptor 2 pathway drives streptococcal cell wall-induced joint inflammation: critical role of myeloid differentiation factor 88. J Immunol. 2003;171(11):6145–53. doi: 10.4049/jimmunol.171.11.6145 14634130.

10. Abdollahi-Roodsaz S, Joosten LA, Roelofs MF, Radstake TR, Matera G, Popa C, et al. Inhibition of Toll-like receptor 4 breaks the inflammatory loop in autoimmune destructive arthritis. Arthritis Rheum. 2007;56(9):2957–67. doi: 10.1002/art.22848 17763416.

11. Kelly PN, Romero DL, Yang Y, Shaffer AL 3rd, Chaudhary D, Robinson S, et al. Selective interleukin-1 receptor-associated kinase 4 inhibitors for the treatment of autoimmune disorders and lymphoid malignancy. J Exp Med. 2015;212(13):2189–201. doi: 10.1084/jem.20151074 26621451; PubMed Central PMCID: PMC4689168.

12. Brentano F, Kyburz D, Gay S. Toll-like receptors and rheumatoid arthritis. Methods Mol Biol. 2009;517:329–43. Epub 2009/04/21. doi: 10.1007/978-1-59745-541-1_20 19378024.

13. Sacre SM, Andreakos E, Kiriakidis S, Amjadi P, Lundberg A, Giddins G, et al. The Toll-like receptor adaptor proteins MyD88 and Mal/TIRAP contribute to the inflammatory and destructive processes in a human model of rheumatoid arthritis. Am J Pathol. 2007;170(2):518–25. doi: 10.2353/ajpath.2007.060657 17255320.

14. Sacre SM, Lo A, Gregory B, Simmonds RE, Williams L, Feldmann M, et al. Inhibitors of TLR8 reduce TNF production from human rheumatoid synovial membrane cultures. J Immunol. 2008;181(11):8002–9. Epub 2008/11/20. 181/11/8002 [pii]. doi: 10.4049/jimmunol.181.11.8002 19017992.

15. Clanchy FI, Sacre SM. Modulation of toll-like receptor function has therapeutic potential in autoimmune disease. Expert Opin Biol Ther. 2010;10(12):1703–16. doi: 10.1517/14712598.2010.534080 21039312.

16. Hennessy EJ, Parker AE, O'Neill LA. Targeting Toll-like receptors: emerging therapeutics? Nat Rev Drug Discov. 2010;9(4):293–307. doi: 10.1038/nrd3203 20380038.

17. Park JE, Kim YI, Yi AK. Protein kinase D1 is essential for MyD88-dependent TLR signaling pathway. J Immunol. 2009;182(10):6316–27. doi: 10.4049/jimmunol.0804239 19414785

18. Park JE, Kim YI, Yi AK. Protein kinase D1: a new component in TLR9 signaling. J Immunol. 2008;181(3):2044–55. doi: 10.4049/jimmunol.181.3.2044 18641342.

19. Upadhyay K, Park JE, Yoon TW, Halder P, Kim YI, Metcalfe V, et al. Group B Streptococci induce proinflammatory responses via a protein kinase D1-dependent pathway. J Immunol. 2017;198(11):4448–57. doi: 10.4049/jimmunol.1601089 28461572; PubMed Central PMCID: PMC5509061.

20. Kim YI, Park JE, Brand DD, Fitzpatrick EA, Yi AK. Protein kinase D1 is essential for the proinflammatory response induced by hypersensitivity pneumonitis-causing thermophilic actinomycetes Saccharopolyspora rectivirgula. J Immunol. 2010;184(6):3145–56. doi: 10.4049/jimmunol.0903718 20142359.

21. Kim YI, Park JE, Kwon KH, Hong CY, Yi AK. Interleukin-1 receptor-associated kinase 2- and protein kinase D1-dependent regulation of IRAK-monocyte expression by CpG DNA. PLoS One. 2012;7(8):e43970. Epub 2012/08/29. doi: 10.1371/journal.pone.0043970 PONE-D-11-22708 [pii]. 22928050; PubMed Central PMCID: PMC3426515.

22. Tang B, Kim S, Hammond S, Cullins DL, Brand DD, Rosloniec EF, et al. Characterization of T cell phenotype and function in a double transgenic (collagen-specific TCR/HLA-DR1) humanized model of arthritis. Arthritis Res Ther. 2014;16(1):R7. doi: 10.1186/ar4433 24405551; PubMed Central PMCID: PMC3978884.

23. Rosloniec EF, Brand DD, Myers LK, Whittington KB, Gumanovskaya M, Zaller DM, et al. An HLA-DR1 transgene confers susceptibility to collagen-induced arthritis elicited with human type II collagen. J Exp Med. 1997;185(6):1113–22. Epub 1997/03/17. doi: 10.1084/jem.185.6.1113 9091584; PubMed Central PMCID: PMC2196244.

24. Yeo SJ, Yoon JG, Hong SC, Yi AK. CpG DNA induces self and cross-hyporesponsiveness of RAW264.7 cells in response to CpG DNA and lipopolysaccharide: alterations in IL-1 receptor-associated kinase expression. J Immunol. 2003;170(2):1052–61. doi: 10.4049/jimmunol.170.2.1052 12517973.

25. Boissier MC, Chiocchia G, Ronziere MC, Herbage D, Fournier C. Arthritogenicity of minor cartilage collagens (types IX and XI) in mice. Arthritis Rheum. 1990;33(1):1–8. doi: 10.1002/art.1780330101 2302260.

26. Rosloniec EF, Cremer M, Kang AH, Myers LK, Brand DD. Collagen-induced arthritis. Curr Protoc Immunol. 2010;Chapter 15:Unit 15 5 1–25. doi: 10.1002/0471142735.im1505s89 20376842.

27. Sieper J, Kary S, Sorensen H, Alten R, Eggens U, Huge W, et al. Oral type II collagen treatment in early rheumatoid arthritis. A double-blind, placebo-controlled, randomized trial. Arthritis Rheum. 1996;39(1):41–51. doi: 10.1002/art.1780390106 8546737.

28. Brand DD, Latham KA, Rosloniec EF. Collagen-induced arthritis. Nat Protoc. 2007;2(5):1269–75. doi: 10.1038/nprot.2007.173 17546023.

29. Nakasa T, Shibuya H, Nagata Y, Niimoto T, Ochi M. The inhibitory effect of microRNA-146a expression on bone destruction in collagen-induced arthritis. Arthritis Rheum. 2011;63(6):1582–90. doi: 10.1002/art.30321 21425254.

30. Sancho D, Gomez M, Viedma F, Esplugues E, Gordon-Alonso M, Garcia-Lopez MA, et al. CD69 downregulates autoimmune reactivity through active transforming growth factor-beta production in collagen-induced arthritis. J Clin Invest. 2003;112(6):872–82. doi: 10.1172/JCI19112 12975472; PubMed Central PMCID: PMC193672.

31. Cho H, Bhatti FU, Lee S, Brand DD, Yi AK, Hasty KA. In vivo dual fluorescence imaging to detect joint destruction. Artif Organs. 2016;40(10):1009–13. doi: 10.1111/aor.12685 27183538.

32. Cho H, Bhatti FU, Yoon TW, Hasty KA, Stuart JM, Yi AK. Non-invasive dual fluorescence in vivo imaging for detection of macrophage infiltration and matrix metalloproteinase (MMP) activity in inflammatory arthritic joints. Biomed Opt Express. 2016;7(5):1842–52. doi: 10.1364/BOE.7.001842 27231625; PubMed Central PMCID: PMC4871085.

33. Iglesias T, Rozengurt E. Protein kinase D activation by mutations within its pleckstrin homology domain. J Biol Chem. 1998;273(1):410–6. doi: 10.1074/jbc.273.1.410 9417097.

34. Leahy AA, Esfahani SA, Foote AT, Hui CK, Rainbow RS, Nakamura DS, et al. Analysis of the trajectory of osteoarthritis development in a mouse model by serial near-infrared fluorescence imaging of matrix metalloproteinase activities. Arthritis Rheumatol. 2015;67(2):442–53. doi: 10.1002/art.38957 25385707; PubMed Central PMCID: PMC4312249.

35. Bottini N, Firestein GS. Duality of fibroblast-like synoviocytes in RA: passive responders and imprinted aggressors. Nat Rev Rheumatol. 2013;9(1):24–33. doi: 10.1038/nrrheum.2012.190 23147896; PubMed Central PMCID: PMC3970924.

36. Neumann E, Lefevre S, Zimmermann B, Gay S, Muller-Ladner U. Rheumatoid arthritis progression mediated by activated synovial fibroblasts. Trends Mol Med. 2010;16(10):458–68. doi: 10.1016/j.molmed.2010.07.004 20739221.

37. Noss EH, Brenner MB. The role and therapeutic implications of fibroblast-like synoviocytes in inflammation and cartilage erosion in rheumatoid arthritis. Immunol Rev. 2008;223:252–70. doi: 10.1111/j.1600-065X.2008.00648.x 18613841.

38. Filer A. The fibroblast as a therapeutic target in rheumatoid arthritis. Curr Opin Pharmacol. 2013;13(3):413–9. doi: 10.1016/j.coph.2013.02.006 23562164.

39. Jones DS, Jenney AP, Swantek JL, Burke JM, Lauffenburger DA, Sorger PK. Profiling drugs for rheumatoid arthritis that inhibit synovial fibroblast activation. Nat Chem Biol. 2017;13(1):38–45. doi: 10.1038/nchembio.2211 27820799; PubMed Central PMCID: PMC5372219.

40. Bartok B, Firestein GS. Fibroblast-like synoviocytes: key effector cells in rheumatoid arthritis. Immunol Rev. 2010;233(1):233–55. doi: 10.1111/j.0105-2896.2009.00859.x 20193003; PubMed Central PMCID: PMC2913689.

41. Ivison SM, Graham NR, Bernales CQ, Kifayet A, Ng N, Shobab LA, et al. Protein kinase D interaction with TLR5 is required for inflammatory signaling in response to bacterial flagellin. J Immunol. 2007;178(9):5735–43. doi: 10.4049/jimmunol.178.9.5735 17442957.

42. Jeohn GH, Cooper CL, Jang KJ, Liu B, Lee DS, Kim HC, et al. Go6976 inhibits LPS-induced microglial TNFalpha release by suppressing p38 MAP kinase activation. Neuroscience. 2002;114(3):689–97. doi: 10.1016/s0306-4522(02)00356-1 12220570.

43. Gschwendt M, Dieterich S, Rennecke J, Kittstein W, Mueller HJ, Johannes FJ. Inhibition of protein kinase C mu by various inhibitors. Differentiation from protein kinase C isoenzymes. FEBS Lett. 1996;392(2):77–80. doi: 10.1016/0014-5793(96)00785-5 8772178.

44. Boissier MC, Chiocchia G, Bessis N, Hajnal J, Garotta G, Nicoletti F, et al. Biphasic effect of interferon-gamma in murine collagen-induced arthritis. Eur J Immunol. 1995;25(5):1184–90. doi: 10.1002/eji.1830250508 7774621.

45. Brand DD, Marion TN, Myers LK, Rosloniec EF, Watson WC, Stuart JM, et al. Autoantibodies to murine type II collagen in collagen-induced arthritis: a comparison of susceptible and nonsusceptible strains. J Immunol. 1996;157(11):5178–84. 8943430.

46. Mukherjee P, Wu B, Mayton L, Kim SH, Robbins PD, Wooley PH. TNF receptor gene therapy results in suppression of IgG2a anticollagen antibody in collagen induced arthritis. Ann Rheum Dis. 2003;62(8):707–14. doi: 10.1136/ard.62.8.707 12860724; PubMed Central PMCID: PMC1754640.

47. Myers LK, Rosloniec EF, Cremer MA, Kang AH. Collagen-induced arthritis, an animal model of autoimmunity. Life Sci. 1997;61(19):1861–78. doi: 10.1016/s0024-3205(97)00480-3 9364191.

48. Kohn EA, Yoo CJ, Eastman A. The protein kinase C inhibitor Go6976 is a potent inhibitor of DNA damage-induced S and G2 cell cycle checkpoints. Cancer Res. 2003;63(1):31–5. 12517773.

49. Smith J, Tho LM, Xu N, Gillespie DA. The ATM-Chk2 and ATR-Chk1 pathways in DNA damage signaling and cancer. Advances in cancer research. 2010;108:73–112. Epub 2010/11/03. doi: 10.1016/B978-0-12-380888-2.00003-0 21034966.

50. Kurosu T, Nagao T, Wu N, Oshikawa G, Miura O. Inhibition of the PI3K/Akt/GSK3 pathway downstream of BCR/ABL, Jak2-V617F, or FLT3-ITD downregulates DNA damage-induced Chk1 activation as well as G2/M arrest and prominently enhances induction of apoptosis. PLoS One. 2013;8(11):e79478. Epub 2013/11/22. doi: 10.1371/journal.pone.0079478 24260231; PubMed Central PMCID: PMC3832535.

51. Huang SW, Chang SH, Mu SW, Jiang HY, Wang ST, Kao JK, et al. Imiquimod activates p53-dependent apoptosis in a human basal cell carcinoma cell line. J Dermatol Sci. 2016;81(3):182–91. doi: 10.1016/j.jdermsci.2015.12.011 26775629.

52. Jin ZH, Kurosu T, Yamaguchi M, Arai A, Miura O. Hematopoietic cytokines enhance Chk1-dependent G2/M checkpoint activation by etoposide through the Akt/GSK3 pathway to inhibit apoptosis. Oncogene. 2005;24(12):1973–81. Epub 2005/01/28. doi: 10.1038/sj.onc.1208408 15674326.

53. Carroll BL, Pulkoski-Gross MJ, Hannun YA, Obeid LM. CHK1 regulates NF-kappaB signaling upon DNA damage in p53- deficient cells and associated tumor-derived microvesicles. Oncotarget. 2016;7(14):18159–70. Epub 2016/02/28. doi: 10.18632/oncotarget.7566 26921248; PubMed Central PMCID: PMC4951279.

54. Duan S, Tsai Y, Keng P, Chen Y, Lee SO, Chen Y. IL-6 signaling contributes to cisplatin resistance in non-small cell lung cancer via the up-regulation of anti-apoptotic and DNA repair associated molecules. Oncotarget. 2015;6(29):27651–60. Epub 2015/08/28. doi: 10.18632/oncotarget.4753 26313152; PubMed Central PMCID: PMC4695015.

55. Kim KS, Choi KJ, Bae S. Interferon-gamma enhances radiation-induced cell death via downregulation of Chk1. Cancer biology & therapy. 2012;13(11):1018–25. Epub 2012/07/25. doi: 10.4161/cbt.20990 22825336; PubMed Central PMCID: PMC3461808.

56. Harikumar KB, Kunnumakkara AB, Ochi N, Tong Z, Deorukhkar A, Sung B, et al. A novel small-molecule inhibitor of protein kinase D blocks pancreatic cancer growth in vitro and in vivo. Molecular cancer therapeutics. 2010;9(5):1136–46. doi: 10.1158/1535-7163.MCT-09-1145 20442301; PubMed Central PMCID: PMC2905628.

Článek vyšel v časopise


2019 Číslo 12
Nejčtenější tento týden