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Autophagy-based unconventional secretion of HMGB1 by keratinocytes plays a pivotal role in psoriatic skin inflammation.
Autophagy ( IF 14.6 ) Pub Date : 2020-02-16 , DOI: 10.1080/15548627.2020.1725381
Zhen Wang 1 , Hong Zhou 1 , Huaping Zheng 1 , Xikun Zhou 1 , Guobo Shen 1 , Xiu Teng 1 , Xiao Liu 1 , Jun Zhang 1 , Xiaoqiong Wei 1 , Zhonglan Hu 1 , Fanlian Zeng 1 , Yawen Hu 1 , Jing Hu 1 , Xiaoyan Wang 1 , Shuwen Chen 1 , Juan Cheng 1 , Chen Zhang 1 , Yiyue Gui 2 , Song Zou 2 , Yan Hao 1 , Qixiang Zhao 1 , Wenling Wu 1 , Yifan Zhou 1 , Kaijun Cui 2 , Nongyu Huang 1 , Yuquan Wei 1 , Wei Li 3 , Jiong Li 1
Affiliation  

The precise mechanism through which macroautophagy/autophagy affects psoriasis is poorly understood. Here, we found that keratinocyte (KC) autophagy, which was positively correlated with psoriatic severity in patients and mouse models and could be inhibited by mitogen-activated protein kinase (MAPK) family inactivation. The impairment of autophagic flux alleviated psoriasisform inflammation. We also found that an autophagy-based unconventional secretory pathway (autosecretion) dependent on ATG5 (autophagy related 5) and GORASP2 (golgi reassembly stacking protein 2) promoted psoriasiform KC inflammation. Moreover, the alarmin HMGB1 (high mobility group box 1) was more effective than other autosecretory proteins in regulating psoriasiform cutaneous inflammation. HMGB1 neutralization in autophagy-efficient KCs eliminated the differences in psoriasiform inflammation between Krt14+/+-Atg5f/f KCs and Krt14Cre/+-atg5f/f KCs, and conversely, recombinant HMGB1 almost completely restored psoriasiform inflammation in Krt14Cre/+-atg5f/f KCs in vivo. These results suggest that HMGB1-associated autosecretion plays a pivotal role in cutaneous inflammation. Finally, we demonstrated that Krt14Cre/+-hmgb1f/f mice displayed attenuated psoriatic inflammation due to the essential crosstalk between KC-specific HMGB1-associated autosecretion and γδT cells. Thus, this study uncovered a novel autophagy mechanism in psoriasis pathogenesis, and the findings imply the clinical significance of investigating and treating psoriasis.Abbreviations: 3-MA: 3-methyladenine; ACTB: actin beta; AGER: advanced glycosylation end-product specific receptor; Anti-HMGB1: anti-HMGB1 neutralizing antibody; Anti-IL18: anti-IL18 neutralizing antibody; Anti-IL1B: anti-IL1B neutralizing antibody; ATG5: autophagy related 5; BAF: bafilomycin A1; BECN1: beclin 1; CASP1: caspase 1; CCL: C-C motif chemokine ligand; CsA: cyclosporine A; ctrl shRNA: lentivirus harboring shRNA against control; CXCL: C-X-C motif chemokine ligand; DCs: dendritic cells; DMEM: dulbecco's modified Eagle's medium; ELISA: enzyme-linked immunosorbent assay; EM: electron microscopy; FBS: fetal bovine serum; GORASP2 shRNA: lentivirus harboring shRNA against GORASP2; GORASP2/GRASP55: golgi reassembly stacking protein 2; GR1: a composite epitope between LY6 (lymphocyte antigen 6 complex) locus C1 and LY6 locus G6D antigens; H&E: hematoxylin and eosin; HMGB1: high mobility group box 1; HMGB1 shRNA: lentivirus harboring shRNA against HMGB1; IFNG/IFN-γ: interferon gamma; IL17A: interleukin 17A; IL18: interleukin 18; IL1A/IL-1α: interleukin 1 alpha; IL1B/IL-1β: interleukin 1 beta; IL22/IL-22: interleukin 22; IL23A: interleukin 23 subunit alpha; IL23R: interleukin 23 receptor; IMQ: imiquimod; ITGAM/CD11B: integrin subunit alpha M; ITGAX/CD11C: integrin subunit alpha X; IVL: involucrin; KC: keratinocyte; KD: knockdown; KO: knockout; Krt14+/+-Atg5f/f mice: mice bearing an Atg5 flox allele, in which exon 3 of the Atg5 gene is flanked by two loxP sites; Krt14+/+-Hmgb1f/f: mice bearing an Hmgb1 flox allele, in which exon 2 to 4 of the Hmgb1 gene is flanked by two loxP sites; Krt14Cre/+-atg5f/f mice: keratinocyte-specific atg5 knockout mice generated by mating Atg5-floxed mice with mice expressing Cre recombinase under the control of the promoter of Krt4; Krt14Cre/+-hmgb1f/f mice: keratinocyte-specific hmgb1 knockout mice generated by mating Hmgb1-floxed mice with mice expressing Cre recombinase under the control of the promoter of Krt14; Krt14-Vegfa mice: mice expressing 164-amino acid Vegfa splice variant recombinase under the control of promoter of Krt14; LAMP1: lysosomal associated membrane protein 1; LDH: lactate dehydrogenase; LORICRIN: loricrin cornified envelope precursor protein; M5: TNF, IL1A, IL17A, IL22 and OSM in combination; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MAPK: mitogen-activated protein kinase; MKI67: marker of proliferation Ki-67; MTT: thiazolyl blue tetrazolium bromide; NFKB/NF-κB: nuclear factor kappa B; NHEKs: primary normal human epidermal keratinocytes; NS: not significant; OSM: oncostatin M; PASI: psoriasis area and severity index; PtdIns3K: class III phosphatidylinositol 3-kinase; qRT-PCR: quantitative RT-PCR; RELA/p65: RELA proto-oncogene, NF-kB subunit; rHMGB1: recombinant HMGB1; rIL18: recombinant interleukin 18; rIL1B: recombinant interleukin 1 beta; S100A: S100 calcium binding protein A; SQSTM1/p62: sequestosome 1; T17: IL17A-producing T; TCR: T-cell receptor; tcrd KO mice: tcrd (T cell receptor delta chain) knockout mice, which show deficient receptor expression in all adult lymphoid and epithelial organs; TLR: toll-like receptor; TNF/TNF-α: tumor necrosis factor; WOR: wortmannin; WT: wild-type; γδT17 cells: IL17A-producing γδ T cells.

中文翻译:

角质形成细胞基于自噬的非常规分泌 HMGB1 在银屑病皮肤炎症中起关键作用。

巨自噬/自噬影响银屑病的确切机制知之甚少。在这里,我们发现角质形成细胞 (KC) 自噬与患者和小鼠模型中的银屑病严重程度呈正相关,并且可以通过丝裂原活化蛋白激酶 (MAPK) 家族失活来抑制。自噬通量的受损减轻了银屑病炎症。我们还发现依赖于 ATG5(自噬相关 5)和 GORASP2(高尔基体重组堆积蛋白 2)的基于自噬的非常规分泌途径(自分泌)促进了银屑病 KC 炎症。此外,alarmin HMGB1(高迁移率族框 1)在调节银屑病样皮肤炎症方面比其他自分泌蛋白更有效。自噬高效 KCs 中的 HMGB1 中和消除了 Krt14+/+-Atg5f/f KCs 和 Krt14Cre/+-atg5f/f KCs 之间银屑病炎症的差异,相反,重组 HMGB1 几乎完全恢复了 Krt14Cre/+-atg5f/f 中的银屑病炎症体内 KCs。这些结果表明 HMGB1 相关的自动分泌在皮肤炎症中起着关键作用。最后,我们证明 Krt14Cre/+-hmgb1f/f 小鼠由于 KC 特异性 HMGB1 相关自分泌和 γδT 细胞之间的基本串扰而表现出减弱的银屑病炎症。因此,本研究揭示了银屑病发病机制中一种新的自噬机制,这些发现对研究和治疗银屑病具有临床意义。缩写:3-MA:3-甲基腺嘌呤;ACTB:肌动蛋白β;年龄:高级糖基化终产物特异性受体;Anti-HMGB1:抗HMGB1中和抗体;Anti-IL18:抗IL18中和抗体;抗 IL1B:抗 IL1B 中和抗体;ATG5:自噬相关5;BAF:巴弗洛霉素A1;BECN1: beclin 1; CASP1:半胱天冬酶1;CCL:CC基序趋化因子配体;CsA:环孢菌素A;ctrl shRNA:慢病毒携带 shRNA 对抗对照;CXCL:CXC基序趋化因子配体;DCs:树突状细胞;DMEM:dulbecco 改良的 Eagle 培养基;ELISA:酶联免疫吸附测定;EM:电子显微镜;FBS:胎牛血清;GORASP2 shRNA:含有针对 GORASP2 的 shRNA 的慢病毒;GORASP2/GRASP55:高尔基体重组堆积蛋白2;GR1:LY6(淋巴细胞抗原6复合物)基因座C1和LY6基因座G6D抗原之间的复合表位;H&E:苏木精和伊红;HMGB1:高机动群箱1个;HMGB1 shRNA:含有针对 HMGB1 的 shRNA 的慢病毒;IFNG/IFN-γ:干扰素γ;IL17A:白细胞介素17A;IL18:白介素 18;IL1A/IL-1α:白介素 1 α;IL1B/IL-1β:白细胞介素 1β;IL22/IL-22:白介素 22;IL23A:白介素 23 亚基 α;IL23R:白介素 23 受体;IMQ:咪喹莫特;ITGAM/CD11B:整合素亚基αM;ITGAX/CD11C:整合素亚基αX;IVL:外皮蛋白;KC:角质形成细胞;KD:击倒;KO:淘汰赛;Krt14+/+-Atg5f/f 小鼠:携带 Atg5 flox 等位基因的小鼠,其中 Atg5 基因的外显子 3 两侧有两个 loxP 位点;Krt14+/+-Hmgb1f/f:携带 Hmgb1 flox 等位基因的小鼠,其中 Hmgb1 基因的外显子 2 至 4 两侧是两个 loxP 位点;Krt14Cre/+-atg5f/f 小鼠:通过将 Atg5-floxed 小鼠与在 Krt4 启动子控制下表达 Cre 重组酶的小鼠交配产生的角质形成细胞特异性 atg5 敲除小鼠;Krt14Cre/+-hmgb1f/f 小鼠:通过将 Hmgb1-floxed 小鼠与在 Krt14 启动子控制下表达 Cre 重组酶的小鼠交配产生的角质形成细胞特异性 hmgb1 敲除小鼠;Krt14-Vegfa 小鼠:在 Krt14 启动子控制下表达 164 个氨基酸的 Vegfa 剪接变异重组酶的小鼠;LAMP1:溶酶体相关膜蛋白 1;LDH:乳酸脱氢酶;LORICRIN:loricrin角化包膜前体蛋白;M5:TNF、IL1A、IL17A、IL22和OSM组合;MAP1LC3/LC3:微管相关蛋白1轻链3;MAPK:丝裂原活化蛋白激酶;MKI67:增殖标记Ki-67;MTT:噻唑蓝四唑溴化物;NFKB/NF-κB:核因子κB;NHEKs:原代正常人表皮角质形成细胞;NS:不显着;OSM:抑癌素 M;PASI:银屑病面积和严重程度指数;PtdIns3K:III类磷脂酰肌醇3-激酶;qRT-PCR:定量RT-PCR;RELA/p65:RELA 原癌基因,NF-kB 亚基;rHMGB1:重组HMGB1;rIL18:重组白介素 18;rIL1B:重组白介素 1 β;S100A:S100钙结合蛋白A;SQSTM1/p62:sequestosome 1;T17:产生 IL17A 的 T;TCR:T细胞受体;tcrd KO 小鼠:tcrd(T 细胞受体 δ 链)敲除小鼠,其在所有成年淋巴和上皮器官中均表现出受体表达缺陷;TLR:toll样受体;TNF/TNF-α:肿瘤坏死因子;WOR:渥曼青霉素;WT:野生型;γδT17 细胞:产生 IL17A 的 γδ T 细胞。不重要; OSM:抑癌素 M;PASI:银屑病面积和严重程度指数;PtdIns3K:III类磷脂酰肌醇3-激酶;qRT-PCR:定量RT-PCR;RELA/p65:RELA 原癌基因,NF-kB 亚基;rHMGB1:重组HMGB1;rIL18:重组白介素 18;rIL1B:重组白介素 1 β;S100A:S100钙结合蛋白A;SQSTM1/p62:sequestosome 1;T17:产生 IL17A 的 T;TCR:T细胞受体;tcrd KO 小鼠:tcrd(T 细胞受体 δ 链)敲除小鼠,其在所有成年淋巴和上皮器官中均表现出受体表达缺陷;TLR:toll样受体;TNF/TNF-α:肿瘤坏死因子;WOR:渥曼青霉素;WT:野生型;γδT17 细胞:产生 IL17A 的 γδ T 细胞。不重要; OSM:抑癌素 M;PASI:银屑病面积和严重程度指数;PtdIns3K:III类磷脂酰肌醇3-激酶;qRT-PCR:定量RT-PCR;RELA/p65:RELA 原癌基因,NF-kB 亚基;rHMGB1:重组HMGB1;rIL18:重组白介素 18;rIL1B:重组白介素 1 β;S100A:S100钙结合蛋白A;SQSTM1/p62:sequestosome 1;T17:产生 IL17A 的 T;TCR:T细胞受体;tcrd KO 小鼠:tcrd(T 细胞受体 δ 链)敲除小鼠,其在所有成年淋巴和上皮器官中均表现出受体表达缺陷;TLR:toll样受体;TNF/TNF-α:肿瘤坏死因子;WOR:渥曼青霉素;WT:野生型;γδT17 细胞:产生 IL17A 的 γδ T 细胞。RELA原癌基因,NF-kB亚基;rHMGB1:重组HMGB1;rIL18:重组白介素 18;rIL1B:重组白介素 1 β;S100A:S100钙结合蛋白A;SQSTM1/p62:sequestosome 1;T17:产生 IL17A 的 T;TCR:T细胞受体;tcrd KO 小鼠:tcrd(T 细胞受体 δ 链)敲除小鼠,其在所有成年淋巴和上皮器官中均表现出受体表达缺陷;TLR:toll样受体;TNF/TNF-α:肿瘤坏死因子;WOR:渥曼青霉素;WT:野生型;γδT17 细胞:产生 IL17A 的 γδ T 细胞。RELA原癌基因,NF-kB亚基;rHMGB1:重组HMGB1;rIL18:重组白介素 18;rIL1B:重组白介素 1 β;S100A:S100钙结合蛋白A;SQSTM1/p62:sequestosome 1;T17:产生 IL17A 的 T;TCR:T细胞受体;tcrd KO 小鼠:tcrd(T 细胞受体 δ 链)敲除小鼠,其在所有成年淋巴和上皮器官中均表现出受体表达缺陷;TLR:toll样受体;TNF/TNF-α:肿瘤坏死因子;WOR:渥曼青霉素;WT:野生型;γδT17 细胞:产生 IL17A 的 γδ T 细胞。在所有成人淋巴和上皮器官中显示出缺陷的受体表达;TLR:toll样受体;TNF/TNF-α:肿瘤坏死因子;WOR:渥曼青霉素;WT:野生型;γδT17 细胞:产生 IL17A 的 γδ T 细胞。在所有成人淋巴和上皮器官中显示出缺陷的受体表达;TLR:toll样受体;TNF/TNF-α:肿瘤坏死因子;WOR:渥曼青霉素;WT:野生型;γδT17 细胞:产生 IL17A 的 γδ T 细胞。
更新日期:2020-02-16
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