Cross-species transcriptional networks in Diabetic Glomerulopathy in mouse and man

Murine models have been valuable instruments in defining the pathogenesis of diabetic nephropathy (DN), but they only partially recapitulate disease manifestations of human DN, limiting their utility . In order to define the molecular similarities and differences between human and murine DN, we performed a cross-species comparison of glomerular transcriptional networks. Glomerular gene expression was profiled in patients with early type 2 DN and in three mouse models (streptozotocin DBA/2 mice, db/db C57BLKS, and eNOS-deficient C57BLKS db/db mice). Species-specific transcriptional networks were generated and compared with a novel network-matching algorithm. Three shared, human-mouse cross-species glomerular transcriptional networks containing 143 (Human-STZ), 97 (Human- db/db), and 162 (Human- eNOS-/- db/db) gene nodes were generated. Shared nodes across all networks reflected established pathogenic mechanisms of diabetic complications, such as elements of JAK-STAT and VEGFR signaling pathways . In addition, novel pathways not formally associated with DN and cross-species gene nodes and pathways unique to each of the human-mouse networks were discovered. The human-mouse shared glomerular transcriptional networks will assist DN researchers in the selection of mouse models most relevant to the human disease process of interest. Moreover, they will allow identification of new pathways shared between mice and humans. We used microarrays to analyze the transcriptome of three different diabetic mouse models Glomerular RNA was extracted using the RNeasy Mini Kit and processed for hybridization on Affymetrix GeneChip Mouse Genome 430 2.0 microarrays.小鼠模型（murine models）在阐明糖尿病肾病（diabetic nephropathy, DN）的发病机制中一直是极具价值的研究工具，但此类模型仅能部分重现人类糖尿病肾病的疾病表型，进而限制了其应用价值。 为明确人类与小鼠糖尿病肾病之间的分子异同，我们开展了跨物种肾小球转录组网络比较研究。我们对早期2型糖尿病肾病患者以及三种小鼠模型——链脲佐菌素（streptozotocin）诱导的DBA/2小鼠、db/db C57BLKS小鼠、eNOS敲除（eNOS-deficient）C57BLKS db/db小鼠——的肾小球基因表达进行了基因表达谱分析。 我们构建了物种特异性转录组网络，并通过一种新型网络匹配算法完成比对。最终生成了三组人-小鼠跨物种共有的肾小球转录组网络，分别包含143个（人-STZ模型）、97个（人-db/db模型）以及162个（人-eNOS敲除db/db模型）基因节点。 所有网络中共有的节点均反映了糖尿病并发症已被证实的致病机制，例如JAK-STAT及VEGFR信号通路的相关元件。此外，我们还发现了此前未与糖尿病肾病建立明确关联的新型通路，以及各个人-小鼠配对网络所特有的跨物种基因节点与通路。 这套人-小鼠共有的肾小球转录组网络将助力糖尿病肾病研究者筛选与目标人类疾病进程最具相关性的小鼠模型。同时，该研究也可用于识别小鼠与人类之间共有的全新通路。 我们采用微阵列（microarrays）技术分析了三种不同糖尿病小鼠模型的转录组，具体步骤为：通过RNeasy Mini Kit提取肾小球RNA，并在Affymetrix GeneChip Mouse Genome 430 2.0微阵列芯片上完成杂交处理。