Regulation of inflammasome activation in diabetic nephropathy

最近的数据表明肾小球细胞表达炎性小体调节因子。此外，肾脏内的非髓系来源细胞有助于糖尿病肾病的病程。然而，支持炎症小体在肾小球细胞中的致病作用的明确证据仍然缺乏。同样，炎性小体在肾小球细胞中的功能后果，包括非典型炎性小体和焦亡的潜在作用，也是未知的。此外，迄今为止，控制肾小球细胞炎症小体活化的机制尚不清楚。根据初步数据，我们假设炎性体激活肾小球细胞，特别是足细胞，有助于糖尿病肾病的肾小球病变。我们认为，与无菌炎症相关的焦亡是糖尿病肾病中细胞死亡的原始形式。根据初步数据，我们假设通过凝血蛋白酶、内质网应激反应、mTOR、自噬和线粒体功能障碍以及ROS生成增加的信号传导控制足细胞中炎症小体的激活。此外，基于初步数据，我们假设炎症小体的持续诱导，可能通过表观遗传机制，有助于代谢记忆，在肾小球中产生促炎微环境。为了评估这些假设，我们将评估以下目标：目标1：定义足细胞中炎性小体激活的细胞自主效应。在这个目标中，我们将使用培养的足细胞，在单一和混合培养中，来定义肾小球滤过屏障细胞中炎症小体调节因子的细胞自主功能。这些研究将与针对炎性小体调节因子失活的小鼠体内研究相结合。目标2：明确凝血蛋白酶调节糖尿病肾病炎性体激活的机制。在这个目标中，我们的目标是确定足细胞中控制炎性体激活的机制，重点关注凝血蛋白酶、它们的受体，以及通过p66Shc或ER、mTOR和自噬的潜在细胞内信号传导。体外研究将与体内研究相结合。目标3：表征表观遗传基因诱导与dNP中持续炎性体激活的相关性。使用体内和体外方法的结合，我们旨在确定糖尿病肾病背景下持续炎性体激活的相关性和机制。

Recent data demonstrated that glomerular cells express inflammasome regulators. In addition, non-myeloid derived cells within the kidney contribute to the disease course of diabetic nephropathy. However, unambiguous evidence supporting a pathogenetic role of the inflammasome in glomerular cells is still lacking. Likewise, the functional consequence of the inflammasome in glomerular cells, including the potential role of the non-canonical inflammasome and pyroptosis, are unknown. Furthermore, the mechanisms controlling inflammasome activation in glomerular cells are not known hitherto. Based on preliminary data we hypothesis that inflammasome activation glomerular cells, including in particular podocytes, contributes to glomerulopathy in diabetic nephropathy. We propose that pyroptosis, which is associated with sterile inflammation, is the primordial form of cell death in the context of diabetic nephropathy. Based on preliminary data we postulate that signalling via coagulation proteases, the ER-stress response, mTOR, autophagy, and mitochondrial dysfunction with increased ROS generation controls inflammasome activation in podocytes. Furthermore, based on preliminary data we hypothesize that persistent induction of the inflammasome, potential by epigenetic mechanism, contributes to the metabolic memory, generating a pro-inflammatory micromilieu in the renal glomeruli. To evaluate these hypotheses we will evaluate the following goals: To Goal 1: Defining the cell-autonomous effects of inflammasome activation in podocytes. Within this aim we will use cultured podocytes, in single and mixed cultures, to define the cell-autonomous function of inflammasome regulators in cells of the glomerular filtration barrier. These studies will be combined with in vivo studies using mice with targeted inactivation of inflammasome regulators. To Goal 2: Define the mechanism through which coagulation proteases regulate inflammasome activation in diabetic nephropathy.Within this aim we aim to define the mechanism controlling inflammasome activation in podocytes, focusing on coagulation proteases, their receptors, and potential intracellular signalling via p66Shc or the ER, mTOR, and autophagy. In vitro studies will be combined with in vivo studies. To Goal 3: Characterize the relevance of epigenetic gene induction for persistent inflammasome activation in dNP.Using a combination of in vivo and in vitro approaches we aim to define the relevance and the mechanism of sustained inflammasome activation in the context of diabetic nephropathy.