醫(yī)學(xué)免疫學(xué)課件：DNA甲基化與系統(tǒng)性紅斑狼瘡

1、DNA Methylation & Systematic Lupus ErythematosusDNA甲基化與系統(tǒng)性紅斑狼瘡 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慢性自身免疫性結(jié)締組織疾病累

3、及心臟、關(guān)節(jié)、皮膚、肺、血管等多種器官血清中存在大量的自身抗體(抗核抗體)https:/wiki/File:Symptoms_of_SLE.pngPart One 1.1 Brief introduction and epidemiology發(fā)病率全球范圍：（40-50）/100 000近年來呈上升趨勢女性發(fā)病率高，男女發(fā)病比例為1:9在非洲和亞洲發(fā)病率略高于白種人生存率高，呈上升趨勢 （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臨床表現(xiàn)：面部蝶形紅斑等皮膚癥狀、關(guān)節(jié)疼痛腫大、口鼻潰瘍、脫發(fā)、漿膜炎、蛋白尿或血尿、神經(jīng)癥狀、溶血性貧血、白細(xì)胞血小板降低免疫學(xué)表現(xiàn)：抗雙鏈DNA抗體高、抗Sm抗體、抗磷脂抗體部分C3、 C4、CH50含量低Part One1.3 Pathology點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題補(bǔ)體活化障礙28種疾??；ITGAM, FcR, PRDM1-ATG5,與SLE密切相關(guān)凋亡細(xì)胞清除障礙巨噬細(xì)胞，核碎片抗原呈遞，抗核抗體多克隆B細(xì)胞活性顯著增強(qiáng)更

5、多未成熟B細(xì)胞轉(zhuǎn)化、CD27+/IgD、CD27/IgD 記憶B細(xì)胞T細(xì)胞功能紊亂調(diào)節(jié)B細(xì)胞應(yīng)答、組織損傷細(xì)胞因子Blys，IL-6,IL-17,IL-18,I型干擾素，TNFPart One 1.4 Etiology點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題遺傳因素家族遺傳性，多為多基因共同作用,免疫應(yīng)答相關(guān)的SNP，TNIP1,PRDM1, JAZF1, UHRF1BP1，CD3-9 and PP2Ac10環(huán)境因素紫外線、吸煙、藥物、飲食、感染等雌激素和性別因素女性好發(fā)，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無DNA序列改變，表型發(fā)生可遺傳的改變DNA甲基化胞嘧啶5-C甲基化CpG 二核苷酸區(qū)組蛋白修飾 染色體結(jié)構(gòu)蛋白甲基化和乙?；痬iRNAHedrich C M, Mbert K, Rauen T, et al. DNA methylation in systemic lupus erythematosusJ. Epigenomics, 2016.Part Two2.

7、2 Mechanisms of DNA methylation 點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題DNA甲基化轉(zhuǎn)移酶（DNMT）S-腺苷基甲硫氨酸（SAM）DNMT1 DNMT3(3a, 3b & 3L)甲基維持酶多識別半甲基化的DNA有絲分裂a(bǔ)/b重新甲基化酶；L介導(dǎo)催化活性多識別雙鏈均未有甲基化的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去甲基化被動去甲基化DNMT1活性不足DNA復(fù)制主動去甲基化堿基切除修復(fù)（脫氨酶）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啟動子或者第一個外顯子區(qū)域編碼基因40%以上的啟動子區(qū)域都含有CpG島甲基化修飾抑制基因的轉(zhuǎn)錄阻止轉(zhuǎn)錄因子與CpG的結(jié)合甲基化的結(jié)合蛋白阻止轉(zhuǎn)錄因子與啟動子的結(jié)合（MBDl、MBD2、MBD4、MECPl和MECP2）影響染色質(zhì)的構(gòu)型低甲基化增強(qiáng)相關(guān)基因的轉(zhuǎn)錄表達(dá)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 點擊

10、此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題相關(guān)性證據(jù) CD4+T 細(xì)胞基因組的甲基化總體水平降低 5-azaC處理正常人源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 點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題相關(guān)性證據(jù) CD4+T 細(xì)胞基因組的甲基化總體水平降低 5-azaC處理正常人源CD4+T，自身免疫性 處理后118個基因表達(dá)上調(diào)2倍以上（CD70、IgE,IL-6,LFA-1,CD11a）12個基因表達(dá)水平下調(diào)1/2以下，與SLE病人測序吻合 個體水平，狼瘡樣綜合征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 低甲基化基因及產(chǎn)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點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題ITGAL (CD11a)CD11a和CD18二聚形成LFA-1LFA-1與ICAM-1相結(jié)合,形成TCR-MHC復(fù)合物啟動子區(qū)域低甲基化水平導(dǎo)致CD11a過表達(dá)40%的

14、活躍狼瘡病人過表達(dá)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點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處

15、添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題alu element 低甲基化CD11a過表達(dá)LFA-1水平升高LFA-1更易與ICAM-1結(jié)合T細(xì)胞活化閾值降低T細(xì)胞結(jié)合自身MHCAPC(主要是巨噬細(xì)胞)凋亡Part Three 3.11 CD4+T cellsPRF1（perforin）主要由CD8+T細(xì)胞和NK細(xì)胞表達(dá)貫通孔道，強(qiáng)溶細(xì)胞活性與顆粒酶協(xié)同作用啟動靶細(xì)胞凋亡CD4+T細(xì)胞增強(qiáng)子3側(cè)翼序列、啟動子區(qū)域的去甲基化CD8+T細(xì)胞中沒有發(fā)現(xiàn)顯著地低甲基化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點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題增強(qiáng)子、啟動子去甲基化PRF-1過表達(dá)自身細(xì)胞裂解，碎片巨噬細(xì)胞減少，無法清除Part Three3.11 CD4+T cellsTNFSF7 (CD70)在活化的CD4+、

17、CD8+T細(xì)胞和早期B細(xì)胞中表達(dá)B細(xì)胞表面CD27（TNF家族）的配體TNFSF7 啟動子區(qū)域去甲基化Part Three 3.11 CD4+T cells啟動子區(qū)域去甲基化CD70過表達(dá)CD70/CD27水平升高B細(xì)胞活化、IgG合成增多、細(xì)胞毒T細(xì)胞增多Part Three 3.11 CD4+T cellsTNFSF5(CD40L)TNF超家族成員主要在活化的CD4+T細(xì)胞上表達(dá)B細(xì)胞共刺激分子、募集巨噬細(xì)胞X染色體上的TNFSF5編碼，可能解釋性別差異女性失活染色體高甲基化，另一條低甲基化男性X染色體低甲基化SLE/甲基化抑制劑使非活躍X染色體上基因低甲基化Part Three 3.11

18、 CD4+T cells啟動子區(qū)域去甲基化CD40L過表達(dá)CD40L/CD40水平升高B細(xì)胞活化、發(fā)育、巨噬細(xì)胞招募Part Three3.11 CD4+T cells點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題KIR gene familyNK細(xì)胞、CD4+T細(xì)胞、CD8+T細(xì)胞與靶細(xì)胞MHC I分子相互作用影響NK細(xì)胞溶細(xì)胞活性與IFN-與巨噬細(xì)胞有關(guān)啟動子區(qū)域低甲基化Part Three3.11 CD4+T cellsOther methylation-sensitive genesIL-6，IL-4：激活B細(xì)胞、刺激T細(xì)胞增殖多種效應(yīng)

19、LTRS甲基化抑制：HERVIFI44L,IFIT1, IFIT3, MX1, STAT1 BST2 ：nave T細(xì)胞CD80, HEERC5, IRF7, ISG15, ISG20, ITGAX , PARP12：狼瘡性腎炎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點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題Regulation of DNA methylation in CD4+ T CellsMAPK/ERK通路 PKC活性降低 DNMT1被抑制 PP2Ac, 氧化應(yīng)激miRNA調(diào)節(jié)miR-146a 低表達(dá)miR-30a 過表達(dá)miR-21 與miR-148a轉(zhuǎn)錄因子RFX1DNMT1，HDAC1招募到CD70，CD11a啟動子上減少Gadd45a潛在的主動去甲基化機(jī)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啟動子區(qū)甲基化降低HRES-1 （HERV）低甲基化IL-6使 DNMT1表達(dá)減少Part Three 3.1 Presented in different cell types3.12 B cells CD5 -E1B 啟動子區(qū)甲基化降低C

22、D5 -E1B 過表達(dá)CD5 -E1A 表達(dá)降低BCR閾值降低自身抗體Part Three3.1 Presented in different cell types:3.13 neutrophils點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題LINE-1, Alu, HERV-E and HERV-KIRS序列低甲基化LINE-1低甲基化可能影響臨近基因表達(dá)Part Three3.1 Presented in different cell types3.14 others點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處

23、添加標(biāo)題點擊此處添加標(biāo)題Follicular T helper cells (Tfh)T helper type 2(Th2)Th17 Regulatory T cells (Tregs) cells Part Three 3.2 DNA methylation and etiology遺傳因素環(huán)境因素藥物： ERK通路抑制劑、DNMT抑制劑紫外線：抑制ERK通路來抑制DNMT-1 表達(dá)、Gadd45a感染：細(xì)菌及病毒DNA富含低甲基化CpG誘導(dǎo)；HERV同源飲食：缺乏甲基化供體（葉酸、蛋氨酸和膽堿）雌激素和性別因素 Part Three 3.3 DNA methylation in lupu

24、s diagnosis and therapy檢測CD11a 和 CD70的甲基化水平檢測FoxP3 Treg特定的去甲基化區(qū)的甲基化水平TGP（白芍總苷）：提高ITGAL的甲基化水平MPA (霉酚酸）：抑制CD40L的表達(dá)4. PerspectivesPart FourPart Four點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題點擊此處添加標(biāo)題不同細(xì)胞類型中的DNA甲基化；高通量測序新的基因SLE發(fā)病機(jī)制轉(zhuǎn)化為臨床診斷和治療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