Department of Pathology and Laboratory Medicine, Western University, London, Ontario, Canada.
Ontario Institute for Cancer Research, Toronto, Ontario, Canada.
PLoS One. 2022 Aug 19;17(8):e0270287. doi: 10.1371/journal.pone.0270287. eCollection 2022.
Hyperglycemia-induced transcriptional alterations lead to aberrant synthesis of a large number of pathogenetic molecules leading to functional and structural damage to multiple end organs including the kidneys. Diabetic nephropathy (DN) remains a major cause of end stage renal disease. Multiple epigenetic mechanisms, including alteration of long non-coding RNAs (lncRNAs) may play a significant role mediating the cellular transcriptional activities. We have previously shown that lncRNA ANRIL may mediate diabetes associated molecular, functional and structural abnormalities in DN. Here we explored downstream mechanisms of ANRIL alteration in DN.
We used renal cortical tissues from ANRIL knockout (KO) mice and wild type (WT) mice, with or without streptozotocin (STZ) induced diabetes for RNA sequencing. The differentially expressed genes were identified using edgeR and DESeq2 computational methods. KEGG and Reactome pathway analyses and network analyses using STRING and IPA were subsequently performed.
Diabetic animals showed hyperglycemia, reduced body weight gain, polyuria and increased urinary albumin. Both albuminuria and polyuria were corrected in the KO diabetic mice. RNA analyses showed Diabetes induced alterations of a large number of transcripts in the wild type (WT) animals. ANRIL knockout (KO) prevented a large number of such alterations. The altered transcripts include metabolic pathways, apoptosis, extracellular matrix protein synthesis and degradation, NFKB related pathways, AGE-RAGE interaction pathways etc. ANRIL KO prevented majority of these pathways.
These findings suggest that as ANRIL regulates a large number of molecules of pathogenetic significance, it may potentially be a drug target for DN and other chronic diabetic complications.
高血糖诱导的转录改变导致大量致病分子的异常合成，导致包括肾脏在内的多个终末器官的功能和结构损伤。糖尿病肾病（DN）仍然是终末期肾病的主要原因。多种表观遗传机制，包括长非编码 RNA（lncRNA）的改变，可能在介导细胞转录活性方面发挥重要作用。我们之前已经表明，lncRNA ANRIL 可能介导糖尿病相关的分子、功能和结构异常在 DN 中。在这里，我们探讨了 ANRIL 改变在 DN 中的下游机制。
我们使用 ANRIL 敲除（KO）小鼠和野生型（WT）小鼠的肾皮质组织，有或没有链脲佐菌素（STZ）诱导的糖尿病，进行 RNA 测序。使用 edgeR 和 DESeq2 计算方法鉴定差异表达基因。随后使用 STRING 和 IPA 进行 KEGG 和 Reactome 途径分析和网络分析。
糖尿病动物表现为高血糖、体重增加减少、多尿和尿白蛋白增加。KO 糖尿病小鼠的白蛋白尿和多尿均得到纠正。RNA 分析显示，糖尿病诱导 WT 动物大量转录本发生改变。ANRIL 敲除（KO）阻止了大量这样的改变。改变的转录本包括代谢途径、细胞凋亡、细胞外基质蛋白合成和降解、NFKB 相关途径、AGE-RAGE 相互作用途径等。ANRIL KO 阻止了这些途径的大部分。
这些发现表明，由于 ANRIL 调节大量具有致病意义的分子，它可能成为 DN 和其他慢性糖尿病并发症的潜在药物靶点。
