DAMP activation and orchestration of cytokine storm by the cold shock protein

急性肾损伤是与血压下降相关的疾病的常见后遗症，如败血症或缺氧。死亡细胞和激活的组织驻留细胞引发免疫反应，随后出现器官衰竭，这是一个复杂的混合过程。本申请的重点是阐明共同的潜在机制和细胞反应。在脓毒症中，细菌释放的可溶性因子进入血液，作为病原体相关分子模式（PAMP）循环，并引发局部和全身炎症反应。PAMP被模式识别受体（PRR）识别并激活NF-κ B信号传导，导致损伤相关分子模式（DAMP）的表达增加。DAMP在炎性小体活化、焦亡、坏死凋亡或通过外来体后从核、线粒体或细胞质区室释放。DAMP启动并维持与全身炎症、器官损伤和细胞死亡相关的非感染性炎症反应。Y盒结合蛋白-1（YB-1）是局部和全身炎症反应的介导剂。炎症刺激，如脂多糖和缺氧，触发细胞内上调以及YB 1分泌.用LPS攻击野生型和全身Ybx 1敲除小鼠（Ybx 1DeltaRosaCreERT）。虽然大多数（70%）野生型小鼠死亡，但75%的Ybx 1敲除小鼠存活。炎症环境的变化表明，YB-1不仅存在于分泌组中，而且还调节其组成。这些发现构成了本工作计划的基础，其中将研究YB-1在DAMP激活和细胞因子风暴的编排中的作用。以下问题将得到解决：（i）哪些DAMP是由YB-1以细胞特异性方式调节的？将对野生型和Ybx 1基因敲除小鼠的不同细胞类型中的分泌组进行比较研究。(ii)在体外模型中，YB-1对DAMP依赖的细胞间通讯有何贡献？(iii)在缺血/再灌注模型中，YB-1与损伤持续和保护作用的体内相关性是什么？将产生具有YB-1野生型和YB-1缺陷型细胞的嵌合骨髓动物。将分析分泌组数据的细胞特异性保护因子以及损伤标志物。这些数据将用于设计治疗策略。设想了两种方法：1.通过抑制YB-1活性阻断LPS和缺氧诱导的损伤标志物（DAMPs，细胞因子）;引入已识别的保护因素作为干预策略。总之，该项目提案将扩大我们对炎症分泌体的理解，重点是冷休克蛋白家族的中枢作用分子。研究结果将转化为干预策略，以改善急性肾损伤和启动组织再生。

Acute kidney injury is a common sequela of diseases associated with blood pressure decrements, such as septicemia or hypoxia. Dead cells and activated tissue resident cells trigger an immune response with subsequent organ failure that represents a complex intermingled process. The focus of this application is to elucidate common underlying mechanisms and cell responses. In sepsis, soluble factors released by bacteria into the blood, circulate as pathogen-associated molecular patterns (PAMPs) and incite local and systemic inflammatory responses. PAMPs are recognized by pattern recognizing receptors (PRRs) and activate NF-kB signaling, leading to increased expression of damage-associated molecular patterns (DAMPs). DAMPs are released from the nuclear, mitochondrial, or cytoplasmic compartments following inflammasome activation, pyroptosis, necroptosis, or by exosomes. DAMPs initiate and sustain a noninfectious inflammatory response that is associated with systemic inflammation, organ damage, and cell death. Y-box binding protein-1 (YB-1) is a mediator of local and systemic inflammatory responses. Inflammatory stimuli, such as LPS and hypoxia, trigger intracellular upregulation as well as YB 1 secretion. Wild type and whole body Ybx1 knockout mice (Ybx1DeltaRosaCreERT) were challenged with LPS. While the majority (70%) of wild type mice die, 75% of Ybx1 knockout mice survive. Changes in the inflammatory milieu show that YB-1 is not only present in the secretome but also regulates the composition. These findings form the basis for the present work program, in which the role of YB-1 in DAMP activation and orchestration of cytokine storm will be investigated. The following questions will be addressed: (i) Which DAMPs are regulated by YB-1 in a cell-specific manner? Comparative studies of secretomes in different cell types of wild type and Ybx1 knockout mice will be performed. (ii) What is the contribution of YB-1 for DAMP-dependent cell-cell communication in in vitro models? (iii) What is the in vivo relevance of YB-1 for damage perpetuation and protective effects in an ischemia/reperfusion model. Chimeric bone marrow animals with YB-1 wild type and YB-1-deficient cells will be generated. The secretome data will be analyzed for cell-specific protective factors as well as damage markers. These data will be used to design therapeutic strategies. Two approaches are envisioned: 1. Blockade of LPS and hypoxia-induced damage markers (DAMPs, cytokines) by inhibiting YB-1 activities and 2. To introduce the identified protective factors as an intervention strategy. In summary, the project proposal will expand our understanding of inflammatory secretomes, with a focus on a centrally acting molecule of the cold shock protein family. The results will be translated into intervention strategies to ameliorate acute kidney injury and initiate tissue regeneration.