title: A novel role for histone deacetylase 6 in the regulation of the tolerogenic STAT3/IL-10 pathway in APCs.
abstract: APCs are critical in T cell activation and in the induction of T cell tolerance. Epigenetic modifications of specific genes in the APC play a key role in this process, and among them histone deacetylases (HDACs) have emerged as key participants. HDAC6, one of the members of this family of enzymes, has been shown to be involved in regulation of inflammatory and immune responses. In this study, to our knowledge we show for the first time that genetic or pharmacologic disruption of HDAC6 in macrophages and dendritic cells results in diminished production of the immunosuppressive cytokine IL-10 and induction of inflammatory APCs that effectively activate Ag-specific naive T cells and restore the responsiveness of anergic CD4(+) T cells. Mechanistically, we have found that HDAC6 forms a previously unknown molecular complex with STAT3, association that was detected in both the cytoplasmic and nuclear compartments of the APC. By using HDAC6 recombinant mutants we identified the domain comprising amino acids 503-840 as being required for HDAC6 interaction with STAT3. Furthermore, by re-chromatin immunoprecipitation we confirmed that HDAC6 and STAT3 are both recruited to the same DNA sequence within the Il10 gene promoter. Of note, disruption of this complex by knocking down HDAC6 resulted in decreased STAT3 phosphorylation--but no changes in STAT3 acetylation--as well as diminished recruitment of STAT3 to the Il10 gene promoter region. The additional demonstration that a selective HDAC6 inhibitor disrupts this STAT3/IL-10 tolerogenic axis points to HDAC6 as a novel molecular target in APCs to overcome immune tolerance and tips the balance toward T cell immunity.
authors.affiliation: Department of Immunology and Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612;
authors.forename: Fengdong
authors.initials: F
authors.lastname: Cheng
authors.affiliation: Department of Immunology and Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612;
authors.forename: Maritza
authors.initials: M
authors.lastname: Lienlaf
authors.affiliation: Department of Immunology and Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612;
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: Wang
authors.affiliation: Department of Immunology and Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612;
authors.forename: Patricio
authors.initials: P
authors.lastname: Perez-Villarroel
authors.affiliation: All Children's Research Institute, All Children's Hospital-Johns Hopkins Medicine, St. Petersburg, FL 33701; Division of Pediatric Oncology, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
authors.forename: Calvin
authors.initials: C
authors.lastname: Lee
authors.affiliation: Department of Immunology and Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612;
authors.forename: Karrune
authors.initials: K
authors.lastname: Woan
authors.affiliation: Department of Immunology and Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612;
authors.forename: Jennifer
authors.initials: J
authors.lastname: Rock-Klotz
authors.affiliation: Department of Immunology and Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612;
authors.forename: Eva
authors.initials: E
authors.lastname: Sahakian
authors.affiliation: Department of Immunology and Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612;
authors.forename: David
authors.initials: D
authors.lastname: Woods
authors.affiliation: Department of Immunology and Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612;
authors.forename: Javier
authors.initials: J
authors.lastname: Pinilla-Ibarz
authors.affiliation: Drug Discovery Program, Department of Medicinal Chemistry and Pharmacognosy, University of Illinois, Chicago, IL 60612;
authors.forename: Jay
authors.initials: J
authors.lastname: Kalin
authors.affiliation: Department of Immunology and Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612;
authors.forename: Jianguo
authors.initials: J
authors.lastname: Tao
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104; and.
authors.forename: Wayne
authors.initials: W
authors.lastname: Hancock
authors.affiliation: Drug Discovery Program, Department of Medicinal Chemistry and Pharmacognosy, University of Illinois, Chicago, IL 60612;
authors.forename: Alan
authors.initials: A
authors.lastname: Kozikowski
authors.affiliation: Department of Molecular Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612.
authors.forename: Edward
authors.initials: E
authors.lastname: Seto
authors.affiliation: Department of Immunology and Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612; Alejandro.Villagra@moffitt.org Eduardo.Sotomayor@moffitt.org.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Villagra
authors.affiliation: Department of Immunology and Malignant Hematology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612; Alejandro.Villagra@moffitt.org Eduardo.Sotomayor@moffitt.org.
authors.forename: Eduardo M
authors.initials: EM
authors.lastname: Sotomayor
mesh_terms: D000107:Acetylation / Q000187:drug effects; D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D047369:Chromatin Immunoprecipitation; D003713:Dendritic Cells / Q000201:enzymology / Q000276:immunology; D015870:Gene Expression; D005786:Gene Expression Regulation; D000073864:Histone Deacetylase 6; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006655:Histone Deacetylases / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D007108:Immune Tolerance; D007249:Inflammation / Q000276:immunology; D016753:Interleukin-10 / Q000096:biosynthesis / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology; D008264:Macrophages / Q000201:enzymology / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009154:Mutation; D009842:Oligopeptides / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects / Q000235:genetics; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D034741:RNA, Small Interfering; D011994:Recombinant Proteins / Q000235:genetics; D050796:STAT3 Transcription Factor / Q000737:chemistry / Q000276:immunology*; D014158:Transcription, Genetic
chemical_list: D056572:Histone Deacetylase Inhibitors; C556062:IL10 protein, mouse; D009842:Oligopeptides; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D050796:STAT3 Transcription Factor; C494087:Stat3 protein, mouse; C510756:tubulysin A; D016753:Interleukin-10; C499033:Hdac6 protein, mouse; D000073864:Histone Deacetylase 6; D006655:Histone Deacetylases
references.citation: J Clin Invest. 2013 Dec;123(12):5212-30
references.pmid: 24200693
references.citation: Immunol Cell Biol. 1998 Feb;76(1):34-40
references.pmid: 9553774
references.citation: J Biol Chem. 2006 Aug 11;281(32):22695-706
references.pmid: 16772287
references.citation: Oncogene. 2007 Aug 13;26(37):5420-32
references.pmid: 17694083
references.citation: Hepatology. 2012 Aug;56(2):644-57
references.pmid: 22392728
references.citation: Mol Immunol. 2014 Jul;60(1):44-53
references.pmid: 24747960
references.citation: J Exp Med. 1994 Jul 1;180(1):25-34
references.pmid: 8006585
references.citation: J Immunol. 2007 Mar 1;178(5):2623-9
references.pmid: 17312100
references.citation: Nucleic Acids Res. 2008 Aug;36(13):4510-20
references.pmid: 18611949
references.citation: Nucleic Acids Res. 2001 May 1;29(9):e45
references.pmid: 11328886
references.citation: J Immunol. 2003 Mar 15;170(6):3263-72
references.pmid: 12626585
references.citation: Nat Rev Cancer. 2006 Jan;6(1):38-51
references.pmid: 16397526
references.citation: Nature. 2007 Jun 21;447(7147):972-8
references.pmid: 17538624
references.citation: Immunity. 2008 Apr;28(4):546-58
references.pmid: 18387831
references.citation: J Biol Chem. 2009 Oct 30;284(44):30264-74
references.pmid: 19744931
references.citation: J Biol Chem. 2000 Apr 14;275(15):10887-92
references.pmid: 10753885
references.citation: Nat Rev Immunol. 2010 Mar;10(3):170-81
references.pmid: 20154735
references.citation: Drugs Today (Barc). 2009 Nov;45(11):787-95
references.pmid: 20126671
references.citation: J Am Chem Soc. 2010 Aug 11;132(31):10842-6
references.pmid: 20614936
references.citation: Biochem Biophys Res Commun. 2009 Feb 6;379(2):616-20
references.pmid: 19121623
references.citation: Immunol Cell Biol. 2012 Jan;90(1):55-65
references.pmid: 22105512
references.citation: Nature. 1998 Dec 10;396(6711):594-8
references.pmid: 9859997
references.citation: Biochem J. 2003 Mar 15;370(Pt 3):737-49
references.pmid: 12429021
references.citation: Eur J Immunol. 2007 Nov;37(11):3164-75
references.pmid: 17948265
references.citation: J Biomed Biotechnol. 2011;2011:875824
references.pmid: 21076528
references.citation: J Leukoc Biol. 2010 Jun;87(6):1103-14
references.pmid: 20200406
references.citation: J Med Chem. 2012 Nov 26;55(22):9891-9
references.pmid: 23009203
references.citation: Proc Natl Acad Sci U S A. 1998 Feb 3;95(3):1178-83
references.pmid: 9448305
references.citation: Immunity. 2008 Apr;28(4):468-76
references.pmid: 18400189
references.citation: Nat Rev Mol Cell Biol. 2008 Mar;9(3):206-18
references.pmid: 18292778
references.citation: Proc Natl Acad Sci U S A. 2013 Jun 11;110(24):9806-11
references.pmid: 23720316
references.citation: Nat Biotechnol. 2007 Jan;25(1):84-90
references.pmid: 17211407
references.citation: Mol Cell Biol. 2011 May;31(10):2066-78
references.pmid: 21444725
references.citation: J Leukoc Biol. 2005 Jan;77(1):3-15
references.pmid: 15522916
references.citation: Nat Immunol. 2009 Jan;10(1):13-4
references.pmid: 19088734
references.citation: Trends Cell Biol. 2008 Jun;18(6):291-7
references.pmid: 18472263
references.citation: Nat Rev Immunol. 2009 Oct;9(10):692-703
references.pmid: 19859064
references.citation: Nat Immunol. 2009 Jan;10(1):92-100
references.pmid: 19011628
references.citation: Mol Cell Biol. 2008 Mar;28(5):1688-701
references.pmid: 18180281
references.citation: Nat Med. 1999 Jul;5(7):780-7
references.pmid: 10395323
references.citation: J Biol Chem. 2004 Feb 27;279(9):7685-91
references.pmid: 14670976
references.citation: Immunity. 2003 May;18(5):605-17
references.pmid: 12753738
references.citation: J Med Chem. 2008 Aug 14;51(15):4370-3
references.pmid: 18642892
references.citation: Curr Biol. 2000 Jun 15;10(12):747-9
references.pmid: 10873806
references.citation: J Immunol. 2011 Apr 1;186(7):3986-96
references.pmid: 21368229
references.citation: Blood. 2009 Oct 1;114(14):2926-35
references.pmid: 19641186
references.citation: Cell. 1997 Aug 22;90(4):595-606
references.pmid: 9288740
references.citation: FASEB J. 2010 Aug;24(8):2869-80
references.pmid: 20354139
references.citation: Blood. 2007 Oct 1;110(7):2528-36
references.pmid: 17609432
references.citation: J Immunol. 1991 Oct 15;147(8):2816-23
references.pmid: 1918995
references.citation: J Biol Chem. 2012 Aug 17;287(34):29168-74
references.pmid: 22778253
references.citation: Immunity. 2004 Apr;20(4):417-28
references.pmid: 15084271
references.citation: J Biol Chem. 2005 Mar 25;280(12):11528-34
references.pmid: 15649887
references.citation: Gene. 2005 Dec 19;363:15-23
references.pmid: 16289629
references.citation: Nat Immunol. 2009 Nov;10(11):1178-84
references.pmid: 19783988
references.citation: Nat Med. 1996 Oct;2(10):1096-103
references.pmid: 8837607
references.citation: Science. 2005 Jan 14;307(5707):269-73
references.pmid: 15653507
references.citation: Mol Cell. 2007 Jul 20;27(2):197-213
references.pmid: 17643370
references.citation: Nature. 2002 May 23;417(6887):455-8
references.pmid: 12024216
references.citation: Clin Cancer Res. 2007 Apr 15;13(8):2318-22
references.pmid: 17438089
references.citation: Immunity. 2003 Sep;19(3):425-36
references.pmid: 14499117
references.citation: J Mol Biol. 2010 Jul 16;400(3):284-94
references.pmid: 20493879