医学免疫学课件：DNA甲基化与系统性红斑狼疮

1、DNA Methylation & Systematic Lupus ErythematosusDNA甲基化与系统性红斑狼疮 Content1. Introduction to Systematic Lupus Erythematosus (SLE) 1.1 Brief introduction and epidemiology 1.2 Manifestation 1.3 Pathology 1.4 Etiology2. DNA methylation 2.1 Epigenetics & DNA methylation 2.2 Mechanisms of DNA methylation 2.3

2、 Effects on gene expression3. DNA methylation in SLE 3.1 Presented in different cell types 3.2 Presented in signaling pathway 3.3 DNA methylation in lupus diagnosis and therapy4. Perspectives1. Systematic Lupus Erythematosus (SLE)Part ONEPart One 1.1 Brief introduction and epidemiology慢性自身免疫性结缔组织疾病累

3、及心脏、关节、皮肤、肺、血管等多种器官血清中存在大量的自身抗体(抗核抗体)https:/wiki/File:Symptoms_of_SLE.pngPart One 1.1 Brief introduction and epidemiology发病率全球范围：（40-50）/100 000近年来呈上升趋势女性发病率高，男女发病比例为1:9在非洲和亚洲发病率略高于白种人生存率高，呈上升趋势 （Euro-lupus,92% ；Toronto,12.6%-3.46%）PRINCIPLES AND METHODS FOR ASSESSING AUTOIMMUNITY ASSOCIATED WITH EX

4、POSURE TO CHEMICALSPart One 1.2 Manifestation临床表现：面部蝶形红斑等皮肤症状、关节疼痛肿大、口鼻溃疡、脱发、浆膜炎、蛋白尿或血尿、神经症状、溶血性贫血、白细胞血小板降低免疫学表现：抗双链DNA抗体高、抗Sm抗体、抗磷脂抗体部分C3、 C4、CH50含量低Part One1.3 Pathology点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题补体活化障碍28种疾病；ITGAM, FcR, PRDM1-ATG5,与SLE密切相关凋亡细胞清除障碍巨噬细胞，核碎片抗原呈递，抗核抗体多克隆B细胞活性显著增强更

5、多未成熟B细胞转化、CD27+/IgD、CD27/IgD 记忆B细胞T细胞功能紊乱调节B细胞应答、组织损伤细胞因子Blys，IL-6,IL-17,IL-18,I型干扰素，TNFPart One 1.4 Etiology点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题遗传因素家族遗传性，多为多基因共同作用,免疫应答相关的SNP，TNIP1,PRDM1, JAZF1, UHRF1BP1，CD3-9 and PP2Ac10环境因素紫外线、吸烟、药物、饮食、感染等雌激素和性别因素女性好发，妊娠Tsokos G C. Systemic lupus eryt

6、hematosusJ. N Engl J Med, 2011,365(22):2110-2121.2. DNA methylationPart TwoPart Two2.1 Epigenetics & DNA methylation无DNA序列改变，表型发生可遗传的改变DNA甲基化胞嘧啶5-C甲基化CpG 二核苷酸区组蛋白修饰 染色体结构蛋白甲基化和乙酰化miRNAHedrich C M, Mbert K, Rauen T, et al. DNA methylation in systemic lupus erythematosusJ. Epigenomics, 2016.Part Two2.

7、2 Mechanisms of DNA methylation 点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题DNA甲基化转移酶（DNMT）S-腺苷基甲硫氨酸（SAM）DNMT1 DNMT3(3a, 3b & 3L)甲基维持酶多识别半甲基化的DNA有丝分裂a/b重新甲基化酶；L介导催化活性多识别双链均未有甲基化的DNA胚胎分化Jeltsch, A., and Jurkowska, R.Z. (2014). New concepts in DNA methylation. Trends in biochemical sciences 39, 31

8、0-318.Part Two2.2 Mechanisms of DNA methylationDNA去甲基化被动去甲基化DNMT1活性不足DNA复制主动去甲基化碱基切除修复（脱氨酶）氧化去甲基化水解去甲基化Part Two2.3 Effects on gene expression非甲基化的CPG序列多分布在基因的启动子或者第一个外显子区域编码基因40%以上的启动子区域都含有CpG岛甲基化修饰抑制基因的转录阻止转录因子与CpG的结合甲基化的结合蛋白阻止转录因子与启动子的结合（MBDl、MBD2、MBD4、MECPl和MECP2）影响染色质的构型低甲基化增强相关基因的转录表达3. DNA met

9、hylation in SLEPart ThreePart Three DNA methylation in SLE3.1 Presented in different cell types3.11 CD4+T cells3.12 B cells 3.13 neutrophils3.14 others3.2 DNA methylation and etiology3.3 DNA methylation in lupus diagnosis and therapyPart Three 3.1 Presented in different cell types3.11 CD4+T cells 点击

10、此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题相关性证据 CD4+T 细胞基因组的甲基化总体水平降低 5-azaC处理正常人源CD4+T，自身免疫性Richardson B C, Strahler J R, Pivirotto T S, et al. Phenotypic and functional similarities between 5-azacytidine-treated T cells and a T cell subset in patients with active systemic lupus erythematosusJ.

11、 Arthritis Rheum, 1992,35(6):647-662.Part Three 3.1 Presented in different cell types3.11 CD4+T cells 点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题相关性证据 CD4+T 细胞基因组的甲基化总体水平降低 5-azaC处理正常人源CD4+T，自身免疫性 处理后118个基因表达上调2倍以上（CD70、IgE,IL-6,LFA-1,CD11a）12个基因表达水平下调1/2以下，与SLE病人测序吻合 个体水平，狼疮样综合征Part Three 3.1

12、Presented in different cell types3.11 CD4+T cells Richardson B C, Strahler J R, Pivirotto T S, et al. Phenotypic and functional similarities between 5-azacytidine-treated T cells and a T cell subset in patients with active systemic lupus erythematosusJ. Arthritis Rheum, 1992,35(6):647-662.Part Three

13、 3.11 CD4+T cells 低甲基化基因及产物1、ITGAL (CD11a)2、PRF1（perforin）3、TNFSF7 (CD70)4、CD40LG (CD40L)5、KIR gene family6、other methylation-sensitive genesPart Three3.11 CD4+T cells点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题ITGAL (CD11a)CD11a和CD18二聚形成LFA-1LFA-1与ICAM-1相结合,形成TCR-MHC复合物启动子区域低甲基化水平导致CD11a过表达40%的

14、活跃狼疮病人过表达CD11aRichardson B C, Strahler J R, Pivirotto T S, et al. Phenotypic and functional similarities between 5-azacytidine-treated T cells and a T cell subset in patients with active systemic lupus erythematosusJ. Arthritis Rheum, 1992,35(6):647-662.Part Three3.11 CD4+T cells点击此处添加标题点击此处添加标题点击此处

15、添加标题点击此处添加标题点击此处添加标题点击此处添加标题alu element 低甲基化CD11a过表达LFA-1水平升高LFA-1更易与ICAM-1结合T细胞活化阈值降低T细胞结合自身MHCAPC(主要是巨噬细胞)凋亡Part Three 3.11 CD4+T cellsPRF1（perforin）主要由CD8+T细胞和NK细胞表达贯通孔道，强溶细胞活性与颗粒酶协同作用启动靶细胞凋亡CD4+T细胞增强子3侧翼序列、启动子区域的去甲基化CD8+T细胞中没有发现显著地低甲基化Part Three 3.11 CD4+T cellsKaplan M J, Lu Q, Wu A, et al. Dem

16、ethylation of promoter regulatory elements contributes to perforin overexpression in CD4+ lupus T cellsJ. J Immunol, 2004,172(6):3652-3661.Part Three3.11 CD4+T cells点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题增强子、启动子去甲基化PRF-1过表达自身细胞裂解，碎片巨噬细胞减少，无法清除Part Three3.11 CD4+T cellsTNFSF7 (CD70)在活化的CD4+、

17、CD8+T细胞和早期B细胞中表达B细胞表面CD27（TNF家族）的配体TNFSF7 启动子区域去甲基化Part Three 3.11 CD4+T cells启动子区域去甲基化CD70过表达CD70/CD27水平升高B细胞活化、IgG合成增多、细胞毒T细胞增多Part Three 3.11 CD4+T cellsTNFSF5(CD40L)TNF超家族成员主要在活化的CD4+T细胞上表达B细胞共刺激分子、募集巨噬细胞X染色体上的TNFSF5编码，可能解释性别差异女性失活染色体高甲基化，另一条低甲基化男性X染色体低甲基化SLE/甲基化抑制剂使非活跃X染色体上基因低甲基化Part Three 3.11

18、 CD4+T cells启动子区域去甲基化CD40L过表达CD40L/CD40水平升高B细胞活化、发育、巨噬细胞招募Part Three3.11 CD4+T cells点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题KIR gene familyNK细胞、CD4+T细胞、CD8+T细胞与靶细胞MHC I分子相互作用影响NK细胞溶细胞活性与IFN-与巨噬细胞有关启动子区域低甲基化Part Three3.11 CD4+T cellsOther methylation-sensitive genesIL-6，IL-4：激活B细胞、刺激T细胞增殖多种效应

19、LTRS甲基化抑制：HERVIFI44L,IFIT1, IFIT3, MX1, STAT1 BST2 ：nave T细胞CD80, HEERC5, IRF7, ISG15, ISG20, ITGAX , PARP12：狼疮性肾炎Coit P, Yalavarthi S, Ognenovski M, et al. Epigenome profiling reveals significant DNA demethylation of interferon signature genes in lupus neutrophilsJ. J Autoimmun, 2015,58:59-66.Part

20、Three3.11 CD4+T cells点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题Regulation of DNA methylation in CD4+ T CellsMAPK/ERK通路 PKC活性降低 DNMT1被抑制 PP2Ac, 氧化应激miRNA调节miR-146a 低表达miR-30a 过表达miR-21 与miR-148a转录因子RFX1DNMT1，HDAC1招募到CD70，CD11a启动子上减少Gadd45a潜在的主动去甲基化机制 Long H, Yin H, Wang L, et al. The critical r

21、ole of epigenetics in systemic lupus erythematosus and autoimmunityJ. Journal of Autoimmunity, 2016,74:118-138.Part Three 3.1 Presented in different cell types3.12 B cells CD5-E1B启动子区甲基化降低HRES-1 （HERV）低甲基化IL-6使 DNMT1表达减少Part Three 3.1 Presented in different cell types3.12 B cells CD5 -E1B 启动子区甲基化降低C

22、D5 -E1B 过表达CD5 -E1A 表达降低BCR阈值降低自身抗体Part Three3.1 Presented in different cell types:3.13 neutrophils点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题LINE-1, Alu, HERV-E and HERV-KIRS序列低甲基化LINE-1低甲基化可能影响临近基因表达Part Three3.1 Presented in different cell types3.14 others点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处

23、添加标题点击此处添加标题Follicular T helper cells (Tfh)T helper type 2(Th2)Th17 Regulatory T cells (Tregs) cells Part Three 3.2 DNA methylation and etiology遗传因素环境因素药物： ERK通路抑制剂、DNMT抑制剂紫外线：抑制ERK通路来抑制DNMT-1 表达、Gadd45a感染：细菌及病毒DNA富含低甲基化CpG诱导；HERV同源饮食：缺乏甲基化供体（叶酸、蛋氨酸和胆碱）雌激素和性别因素 Part Three 3.3 DNA methylation in lupu

24、s diagnosis and therapy检测CD11a 和 CD70的甲基化水平检测FoxP3 Treg特定的去甲基化区的甲基化水平TGP（白芍总苷）：提高ITGAL的甲基化水平MPA (霉酚酸）：抑制CD40L的表达4. PerspectivesPart FourPart Four点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题点击此处添加标题不同细胞类型中的DNA甲基化；高通量测序新的基因SLE发病机制转化为临床诊断和治疗References1 Chen S H, Lv Q L, Hu L, et al. DNA methylation alterat

25、ions in the pathogenesis of lupusJ. Clinical & Experimental Immunology, 2016. 2 Hedrich C M, Mbert K, Rauen T, et al. DNA methylation in systemic lupus erythematosusJ. Epigenomics, 2016. 3 Long H, Yin H, Wang L, et al. The critical role of epigenetics in systemic lupus erythematosus and autoimmunity

26、J. Journal of Autoimmunity, 2016,74:118-138. 4 Sun B, Hu L, Luo Z, et al. DNA methylation perspectives in the pathogenesis of autoimmune diseasesJ. Clinical Immunology, 2016,164:21-27. 5 Yang Y, Tang Q, Zhao M, et al. The effect of mycophenolic acid on epigenetic modifications in lupus CD4+T cellsJ.

27、 Clin Immunol, 2015,158(1):67-76. 6 Coit P, Renauer P, Jeffries M A, et al. Renal involvement in lupus is characterized by unique DNA methylation changes in naive CD4+ T cellsJ. J Autoimmun, 2015,61:29-35. 7 Coit P, Yalavarthi S, Ognenovski M, et al. Epigenome profiling reveals significant DNA demet

28、hylation of interferon signature genes in lupus neutrophilsJ. J Autoimmun, 2015,58:59-66. 8 Lisnevskaia L, Murphy G, Isenberg D. Systemic lupus erythematosusJ. Lancet, 2014,384(9957):1878-1888. 9 Richardson B C, Patel D R. Epigenetics in 2013. DNA methylation and miRNA: key roles in systemic autoimm

29、unityJ. Nat Rev Rheumatol, 2014,10(2):72-74.10 Zhao M, Liu S, Luo S, et al. DNA methylation and mRNA and microRNA expression of SLE CD4+ T cells correlate with disease phenotypeJ. J Autoimmun, 2014,54:127-136.References 11 Zhang Y, Zhao M, Sawalha A H, et al. Impaired DNA methylation and its mechani

30、sms in CD4+T cells of systemic lupus erythematosusJ. Journal of Autoimmunity, 2013,41:92-99.12 Jeffries M A, Sawalha A H. Epigenetics in systemic lupus erythematosus: leading the way for specific therapeutic agentsJ. International Journal of Clinical Rheumatology, 2011,6(4):423-438.13 Tsokos G C. Systemic lupus erythematosusJ. N Engl J Med, 2011,365(22):2110-2121.14 Richardson B C, Strahler J R, Pivirotto T S, et al. Phenotypic and functional similarities between 5-azacytidine-treated T cells and a T cell subset i