Recaptulating transcriptional pathways of human diabetic nephropathy in mice

描述(由申请人提供)： 有效的糖尿病肾病小鼠模型应该复制糖尿病肾病患者的分子变化，而不是简单的病理变化。因此，我们对糖尿病肾病小鼠模型的发展和测试的一般假设是，目前的小鼠模型未能显示出与人类相似的糖尿病肾病，因为它们无法复制进行性糖尿病肾病患者中发生的肾小球和肾小管间质基因表达的变化。复制进展性糖尿病肾病患者的关键转录谱可诱发小鼠进展性糖尿病肾病。我们使用欧洲肾脏cDNA银行(ERCB)生成的人类糖尿病肾病数据将是测试和验证糖尿病并发症动物模型的小鼠模型的关键。我们已经使用ERCB对患有糖尿病肾病的人类进行了初步的转录分析，以确定在人类中可靠地改变了哪些通路，但在小鼠模型中却没有。在人类糖尿病患者的肾小球和肾小管间质中，JAK/STAT通路是一条持续改变的通路，而在小鼠中则不然。实时定量聚合酶链式反应证实，所有JAK成员的表达均增加。鉴于JAK2在介导糖尿病肾病反应中的关键作用，我们将重点放在JAK2上。此外，JAK2被活性氧激活，并与PPAR(信号转导)相互作用，这两种信号都与糖尿病肾病有关。对于我们的两个新模型，我们提出了足细胞和近端小管特异的JAK2转基因db/m C57BLKS小鼠。对于财团中的这些和其他型号，我们建议： 1.确定JAK2//db/db BLKS模型和其他AMDCC模型的肾小球和肾小管间质是否复制了人类的转录变化； 2.确定在JAK2//db/db BLKS模型中是否复制了人类糖尿病肾病的所有病理和病理生理特征； 3.在JAK2转基因模型和其他复制人类转录改变的良好候选模型中，确定JAK2/3抑制剂是否可以防止糖尿病肾病的发生； 4.确定ROS的产生是否促进了肾小球和/或肾小管间质中JAK2的表达，并增强了JAK2下游的信号转导，以及JAK2的表达是否促进了ROS； 5.确定PPAR(激动剂)是否能阻止肾小球和/或肾小管间质中的J.ak2下游效应。 这项研究与糖尿病肾脏疾病的研究和预防直接相关，糖尿病肾脏疾病是美国肾衰竭的主要原因。通过创建和了解发展类似人类的糖尿病肾脏疾病的小鼠模型，我们可以迅速进入预防和治疗这种疾病的策略测试。

DESCRIPTION (provided by applicant): Valid murine models of diabetic nephropathy (DN) should replicate the molecular changes and not simply the pathological alterations of patients with DN. Thus, our general hypothesis for development and testing of murine models of diabetic nephropathy is that Current murine models fail to show human-like DN because they fail to replicate glomerular and tubulointerstital gene expression changes that occur in humans with progressive DN. Replication of the critical transcriptomic profiles of patients with progressive DN should induce progressive DN in mice. Our use of data in human DN generated by the European Renal cDNA Bank (ERCB) will be critical in testing and validating the mouse models of the Animal Models of Diabetic Complications Consortium. We have performed initial transcriptomic analyses of humans with DN using the ERCB to identify pathways which are reliably altered in humans but not in murine models. One pathway that is consistently altered in glomeruli and tubulointerstium in diabetes in humans, but not in mice, is the JAK/STAT pathway. Expression of all JAK members was increased when confirmed with real time PCR analysis. We have focused on JAK2 given its key role in mediating responses implicated in DN. Moreover, JAK2 is activated by reactive oxygen species and interacts with PPAR( signaling, both of which are implicated in DN. For our 2 novel models, we propose podocyte and proximal tubular-specific Jak2 transgenic db/m C57BLKS mice. For these and other models in the Consortium we propose to: 1. Determine whether transcriptional changes in humans are reproduced in the glomerular and tubulointersitial compartments of the Jak2//db/db BLKS models, and other AMDCC models; 2. Determine if all the pathologic and pathophysiologic features of human DN are replicated in the Jak2//db/db BLKS models; 3. Determine if JAK2/3 inhibitors prevent development of DN in the Jak2 transgenic models and other good candidate models that replicate human transcriptomic changes; 4. Determine if ROS production drives JAK2 expression in glomerular and/or tubulointerstitial compartments and enhances downstream JAK2 signaling and whether JAK2 expression promotes ROS; 5. Determine if PPAR( agonists prevent J.ak2 downstream effects in glomerular and/or tubulointerstitial compartments. This research is directly relevant to the study and prevention of diabetic kidney disease, the major cause of kidney failure in the U.S. By creating and understanding a mouse model that develops human-like diabetic kidney disease, we can then move rapidly to tests of strategies to prevent and cure this disease.