Drugs from bugs: YopM - a bacteria-derived anti-inflammatory cell-penetrating peptide for the topical treatment of psoriasis

来自细菌的药物：YopM - 一种细菌衍生的抗炎细胞穿透肽，用于局部治疗牛皮癣

• 批准号: 258026263
• 负责人: Professorin Dr. Karin Loser
• 金额: --
• 依托单位: Institut für Infektiologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/258026263?language=en
• 关键词: Drugs; bugs; YopM; bacteria; derived

During Yersinia enterocolitica infection bacteria translocate different Yersinia outer proteins (Yops) into the host cell cytoplasm through a type III secretion system. YopM, in contrast to other Yops, does not possess any enzymatic activity and it has been shown that after translocation into the host cell, YopM traffics to the nucleus via a vesicle-associated pathway. In the nucleus YopM inhibits the transcription of genes encoding pro-inflammatory cytokines such as TNF-alpha, IL 1beta, IFN-gamma, IL-12, or IL-18. Accordingly, in a mouse model of Y. pestis infection YopM reduced the expression of these pro-inflammatory cytokines in macrophages leading to an increased virulence of bacteria as well as a down-regulated anti-bacterial immunity in the host. However, thus far it is not known how a locally translocated effector protein might be responsible for systemic effects on anti-bacterial immune responses. Interestingly, in previous studies we identified YopM as a cell-penetrating peptide and demonstrated its ability to autonomously enter various mammalian cells including immune and epithelial cells via caveolin-dependent endocytosis followed by endosomal escape and without the help of additional factors. In support of this, treatment of activated primary immune cells with recombinant YopM (rYopM) led to the down-regulation of TNF-alpha, IL-1beta and IFN-gamma indicating that rYopM acts as a self-delivering anti-inflammatory drug and therefore, might be of interest for the treatment of inflammatory disorders. To investigate the anti-inflammatory and therapeutic potential of rYopM in vivo we used the mouse model of imiquimod-induced psoriasis. Usually, immunomodulators, corticosteroids or biologics are used for the treatment of psoriasis. However, many of these drugs are ineffective when applied locally because of their inability to penetrate the epidermal barrier. As rYopM breaches barriers and enters host cells without the requirement of additional factors we topically treated psoriatic mice with a rYopM-containing cream and surprisingly, could show that epicutaneous application of rYopM markedly reduced all characteristic hallmarks of psoriasis like acanthosis or papillomatosis. Now, we intend to analyze the cellular and molecular mechanisms underlying the amelioration of psoriasis by topical rYopM treatment in mouse as well as human skin. With the help of tissue culture models the effect of rYopM on signaling networks will be analyzed, including the skin barrier function and the innate as well as adaptive immune response. Furthermore, we will generate truncated rYopM proteins by site-directed mutagenesis to identify domains required for the penetration of epithelial barriers as well as the anti-inflammatory activity. Thereby, rYopM variants with improved immunomodulatory capacities and increased stability will be determined, which might represent the basis for the development of a novel therapeutic alternative for local immunotherapy.

在小肠结肠炎耶尔森氏菌感染期间，细菌通过III型分泌系统将不同的耶尔森氏菌外部蛋白（Yops）转运到宿主细胞质中。与其他Yop相反，YopM不具有任何酶活性，并且已经显示在易位到宿主细胞中之后，YopM经由囊泡相关途径运输到细胞核。在细胞核中，YopM抑制编码促炎细胞因子如TNF-α、IL 1 β、IFN-γ、IL-12或IL-18的基因的转录。因此，在Y.鼠疫菌感染YopM降低了巨噬细胞中这些促炎细胞因子的表达，导致细菌毒力增加以及宿主中抗细菌免疫力下调。然而，到目前为止，还不知道局部易位的效应蛋白如何可能负责对抗细菌免疫应答的全身作用。有趣的是，在先前的研究中，我们将YopM鉴定为细胞穿透肽，并证明其能够通过小窝蛋白依赖性内吞作用自主进入各种哺乳动物细胞，包括免疫细胞和上皮细胞，随后进行内体逃逸，而无需其他因子的帮助。为了支持这一点，用重组YopM（rYopM）处理活化的原代免疫细胞导致TNF-α、IL-1 β和IFN-γ的下调，表明rYopM充当自递送抗炎药物，因此可能对治疗炎性疾病感兴趣。为了研究rYopM在体内的抗炎和治疗潜力，我们使用咪喹莫特诱导的银屑病的小鼠模型。通常，免疫调节剂、皮质类固醇或生物制剂用于治疗银屑病。然而，这些药物中的许多在局部应用时是无效的，因为它们不能穿透表皮屏障。由于rYopM突破屏障并进入宿主细胞而不需要额外的因子，我们用含有rYopM的乳膏局部治疗银屑病小鼠，并且令人惊讶的是，可以显示rYopM的表皮应用显著减少银屑病的所有特征性标志，如棘皮病或乳头状瘤病。现在，我们打算分析的细胞和分子机制的改善银屑病局部rYopM治疗小鼠以及人类皮肤。在组织培养模型的帮助下，将分析rYopM对信号网络的影响，包括皮肤屏障功能以及先天性和适应性免疫应答。此外，我们将通过定点诱变产生截短的rYopM蛋白，以鉴定穿透上皮屏障以及抗炎活性所需的结构域。因此，将确定具有改善的免疫调节能力和增加的稳定性的rYopM变体，这可能代表开发用于局部免疫疗法的新型治疗替代方案的基础。

项目成果

The tripeptide KdPT ameliorates ongoing psoriasis‐like skin inflammation in murine and human skin

三肽 KdPT 可改善小鼠和人类皮肤中持续存在的类似牛皮癣的皮肤炎症

• DOI: 10.1111/exd.13145
• 发表时间: 2017
• 期刊: Experimental Dermatology
• 影响因子: 3.6
• 作者: Mykicki N;Klenner L;Baumann C;Auriemma M;Sternemann C;Soeberdt M;Elliott GR;Abels C;Luger TA;Loser K
• 通讯作者: Loser K

All in-Multiple parallel strategies for intracellular delivery by bacterial pathogens.

• DOI: 10.1016/j.ijmm.2018.06.007
• 发表时间: 2018-10
• 期刊: International journal of medical microbiology : IJMM
• 作者: C. Rüter;Marie-Luise Lubos;Stefanie Norkowski;M. A. Schmidt

Cutaneous RANK-RANKL Signaling Upregulates CD8-Mediated Antiviral Immunity during Herpes simplex Virus Infection by Preventing Virus-Induced Langerhans Cell Apoptosis.

皮肤 RANK-RANKL 信号传导通过预防病毒诱导的朗格汉斯细胞凋亡来上调单纯疱疹病毒感染期间 CD8 介导的抗病毒免疫

• DOI: 10.1038/jid.2015.225
• 发表时间: 2015
• 期刊: The Journal of investigative dermatology
• 作者: Klenner L;Hafezi W;Clausen BE;Lorentzen EU;Luger TA;Beissert S;Kühn JE;Loser K
• 通讯作者: Loser K

Novel insights into the pathogenesis of psoriasis.

对牛皮癣发病机制的新见解

• DOI: 10.1016/j.clim.2017.07.014
• 发表时间: 2018
• 期刊: Clinical immunology
• 影响因子: 8.6
• 作者: Luger TA;Loser K
• 通讯作者: Loser K

Cell-Penetrating Bacterial Effector Proteins: Better Tools than Targets.

• DOI: 10.1016/j.tibtech.2016.08.002
• 发表时间: 2017-02
• 期刊: Trends in biotechnology
• 影响因子: 17.3
• 作者: C. Rüter;M. A. Schmidt