Functional role of HSP70 expression regulation in the pathogenesis of epilepsy and associated inflammatory processes

过度的炎症信号传导已被证实是促进癫痫发展（=癫痫发生）和癫痫大脑过度兴奋的一个关键机制。考虑到现有的抗癫痫药物无法控制大部分犬类患者的癫痫发作，开发针对癫痫发生或内在疾病严重程度的病理生理机制的预防或疾病改善策略具有特别的意义。热休克蛋白超家族HSP70包括应激诱导型HSP70i，其至少部分通过与toll样受体4 （TLR4）的相互作用介导，作为炎症反应的调节剂。尽管HSP70i的作用有两种，但目前的研究表明，与神经损伤相关的HSP70i上调似乎主要是抗炎作用。根据这一概念，我们将探讨增加HSP70i表达的遗传或药物策略在慢性癫痫模型中发挥有益预防作用的假设。对癫痫发生和癫痫的啮齿动物模型以及犬类患者的死后组织进行免疫组织化学分析，将提供有关HSP70i和HSP70超家族其他成员的疾病相关调节模式的全面信息。这些信息对制定目标战略至关重要。利用转基因HSP70i过表达小鼠，我们将在颞叶癫痫点燃模型中评估该蛋白对癫痫易感性、癫痫进展、小胶质细胞激活和促炎细胞因子产生的影响。在后续的实验中，HSP70i的药理诱导效果将在点火模型中确定。因此，通过在HSP70i和TLR4敲除小鼠中进行平行实验，可以控制药理作用对HSP70i和TLR4的依赖性。我们期望HSP70i靶向实验为未来新型疾病修饰或预防方法的转化开发提供基础。

Excessive inflammatory signaling has been confirmed as one key mechanism contributing to epilepsy development (=epileptogenesis) and to hyperexcitability in the epileptic brain. Considering the fact that available antiepileptic drugs fail to control epileptic seizures in a large subgroup of canine patients, it is of particular interest to develop preventive or disease-modifying strategies targeting the pathophysiological mechanisms of epileptogenesis or of intrinsic disease severity.The heat-shock protein superfamily HSP70 comprises a stress-inducible HSP70i, which at least partly mediated by an interaction with Toll-like receptor 4 (TLR4) acts as modulator of inflammatory responses. Despite a dichotomous role described for HSP70i, the current state-of-knowledge indicates that HSP70i up-regulation associated with neurological insults appears to be predominantly anti-inflammatory. In line with this concept, we will address the hypothesis that genetic or pharmacological strategies increasing HSP70i expression exert beneficial preventive effects in a chronic epilepsy model. Immunhistochemical analysis in tissue from rodent models of epileptogenesis and epilepsy, and in post mortem tissue from canine patients will provide comprehensive information about disease-associated regulation patterns of HSP70i and further members of the HSP70 superfamily. The information is crucial for the development of targeting strategies. Using transgenic HSP70i overexpressing mice we will assess the impact of the protein on seizure susceptibility, seizure progression, microglia activation, and production of pro-inflammatory cytokines in the kindling model of temporal lobe epilepsy. In subsequent experiments, the efficacy of pharmacological induction of HSP70i will be determined in the kindling model. Thereby, the HSP70i- and TLR4-dependency of the pharmacological effect will be controlled by parallel experiments in HSP70i and TLR4 knockout mice. We expect that the HSP70i targeting experiments provide a basis for future translational development of novel disease-modifying or preventive approaches.