Impact of the cytoprotective chaperone HSP25/27 on the structural integrity and autophagic degradation of chlamydial compartments

细胞保护伴侣 HSP25/27 对衣原体区室结构完整性和自噬降解的影响

• 批准号: 260801578
• 负责人: Privatdozent Dr. Michael R. Knittler
• 金额: --
• 依托单位: Institut für Immunologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/260801578?language=en
• 关键词: Impact; cytoprotective; chaperone; HSP25; 27

Chlamydiae are obligate intracellular bacteria that cause sexually transmitted disease, ocular infections, and atypical pneumonia. All known chlamydia species undergo a unique biphasic developmental cycle that takes place within a non-acidified membrane-bound vacuole, termed inclusion. Chlamydiae target the host cytoskeleton to regulate diverse aspects of their intracellular survival such as the structural stability and physiological maintenance of their inclusions. To this end, the bacteria surround their vacuole with a mesh of cytoskeletal filaments, which serve as a scaffold structurally stabilizing the bacterial compartment. As a consequence, the activation of host cell defense mechanisms is limited as leakage of inclusion contents into the cytosol is prevented. We recently demonstrated that chlamydia-infected immune and non-immune cells are able to break down inclusions resulting in the cytosolic release of bacteria. Subsequently, chlamydiae are killed and degraded via autophagy leading to the processing of chlamydial antigens and their display to T cells for immunosurveillance. Moreover, we found that the infection-induced expression of the host cell chaperone HSP25/27, which regulates the remodeling of cytoskeletal filaments, seems to play a critical role in disrupting and degrading chlamydial compartments. Therefore, the detailed elucidation of HSP25/27-mediated processes in chlamydia-infected cells is crucial to understand how inclusions are attacked in host cells and which cellular machinery is used to destroy the bacterial compartments. Our key goal is to unravel the molecular and cellular processes that HSP25/27 impacts and to assess how these processes determine the outcome of chlamydial infections. We will utilize advanced state-of-the-art methods in biochemistry, microscopy, cell biology, and molecular biology to functionally characterize infection-induced HSP25/27 and attempt to identify both host cellular and pathogen-derived HSP25/27 interaction partners. We will also explore how HSP25/27 affects bacterial cytoskeleton effector proteins, and how it disrupts the integrity of persistent and productive inclusions as well as their interaction with the host cytoskeleton. Moreover, we will assess the chlamydial gene expression accompanying inclusion breakdown. Another focus of our work will address the functional cooperation between the disintegration of chlamydial compartments and their subsequent degradation. Here, we will concentrate on NOD1/2 (Nucleotide-Binding Oligomerization Domain 1/2)-pathways and on HMGB1 (High Mobility Group Box 1)-controlled processes of HSP25/27 expression and autophagy. Our studies on cytoprotective HSP25/27 and its effects on bacterial compartments will aid the deeper understanding of the intracellular life cycle of chlamydia and the development of novel therapeutic strategies against chlamydial infections.

衣原体是专性细胞内细菌，可引起性传播疾病、眼部感染和非典型肺炎。所有已知的衣原体物种都经历独特的双相发育周期，该周期发生在非酸化的膜结合液泡内，称为包涵体。衣原体以宿主细胞骨架为目标来调节其细胞内生存的各个方面，例如其内含物的结构稳定性和生理维持。为此，细菌用细胞骨架丝网包围其液泡，细胞骨架丝作为支架在结构上稳定细菌室。因此，宿主细胞防御机制的激活受到限制，因为防止了包涵体内容物渗漏到细胞质中。我们最近证明，衣原体感染的免疫和非免疫细胞能够分解内含物，导致细菌释放到胞质中。随后，衣原体被杀死并通过自噬降解，导致衣原体抗原的加工并将其展示给 T 细胞进行免疫监视。此外，我们发现感染诱导的宿主细胞伴侣HSP25/27的表达（调节细胞骨架丝的重塑）似乎在破坏和降解衣原体区室中发挥着关键作用。因此，详细阐明衣原体感染细胞中 HSP25/27 介导的过程对于了解宿主细胞中内含物如何受到攻击以及使用哪种细胞机制来破坏细菌区室至关重要。我们的主要目标是揭示 HSP25/27 影响的分子和细胞过程，并评估这些过程如何决定衣原体感染的结果。我们将利用生物化学、显微镜、细胞生物学和分子生物学领域最先进的方法来对感染诱导的 HSP25/27 进行功能表征，并尝试鉴定宿主细胞和病原体衍生的 HSP25/27 相互作用伴侣。我们还将探讨 HSP25/27 如何影响细菌细胞骨架效应蛋白，以及它如何破坏持久性和生产性内含物的完整性以及它们与宿主细胞骨架的相互作用。此外，我们将评估伴随包涵体分解的衣原体基因表达。我们工作的另一个重点将解决衣原体区室的崩解与其随后的降解之间的功能合作。在这里，我们将重点关注 NOD1/2（核苷酸结合寡聚化结构域 1/2）途径以及 HMGB1（高迁移率组框 1）控制的 HSP25/27 表达和自噬过程。我们对细胞保护性 HSP25/27 及其对细菌区室影响的研究将有助于更深入地了解衣原体的细胞内生命周期，并开发针对衣原体感染的新型治疗策略。