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（DN）小鼠模型应复制DN患者的分子变化，而不仅仅是病理学改变。因此，我们对糖尿病肾病小鼠模型的开发和测试的一般假设是，目前的小鼠模型未能显示人样DN，因为它们未能复制肾小球和肾小管上皮细胞基因表达的变化，发生在人类进行性DN。进行性DN患者的关键转录组学特征的复制应该在小鼠中诱导进行性DN。我们使用欧洲肾脏cDNA库（ERCB）产生的人类DN数据将在测试和验证糖尿病并发症动物模型联盟的小鼠模型中发挥关键作用。我们使用ERCB对DN患者进行了初步的转录组学分析，以确定在人类中可靠改变但在小鼠模型中不改变的途径。JAK/STAT通路是人类糖尿病患者肾小球和肾小管中持续改变的通路，但在小鼠中没有改变。当用真实的时间PCR分析证实时，所有JAK成员的表达增加。鉴于JAK 2在介导DN相关反应中的关键作用，我们将重点放在JAK 2上。此外，JAK 2被活性氧物质激活并与PPAR（信号传导）相互作用，这两者都与DN有关。对于我们的2种新模型，我们提出了足细胞和近端小管特异性Jak 2转基因db/mC 57 BLKS小鼠。对于联盟中的这些和其他模型，我们建议： 1.确定人类的转录变化是否在Jak 2//db/db BLKS模型和其他AMDCC模型的肾小球和肾小管间质区室中再现; 2.确定人DN的所有病理和病理生理学特征是否在Jak 2//db/db BLKS模型中复制; 3.确定JAK 2/3抑制剂是否能在Jak 2转基因模型和其他复制人类转录组变化的良好候选模型中预防DN的发展; 4.确定ROS产生是否驱动肾小球和/或肾小管间质区室中的JAK 2表达并增强下游JAK 2信号传导，以及JAK 2表达是否促进ROS; 5.确定PPAR激动剂是否阻止肾小球和/或肾小管间质隔室中的J.ak2下游效应。 这项研究与糖尿病肾病的研究和预防直接相关，糖尿病肾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糖尿病肾病的小鼠模型，我们可以快速测试预防和治疗这种疾病的策略。

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.