Sahoo Biswajit, Mishra Deendayal Das, Tiwari Swasti
Department of Molecular Medicine & Biotechnology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, 226014, India.
Noncoding RNA Res. 2024 Nov 19;11:60-72. doi: 10.1016/j.ncrna.2024.11.003. eCollection 2025 Apr.
Diabetic kidney disease (DKD), a.k.a diabetic nephropathy, is a leading cause of end-stage renal disease. However, in a fair percentage of patients with type-2 diabetes, renal involvement also occurs due to non-diabetic reasons (non-diabetic kidney disease, NDKD). In this study, we identified miRNA-mRNA regulatory networks specific to human DKD pathogenesis. miRNA profiling of the renal biopsy from cases (DKD, n = 5), disease controls (T2DM with NDKD, n = 6), and non-diabetic, non-CKD controls (patients undergoing nephrectomy for renal cancer, n = 3) revealed 68 DKD-specific miRNA regulation. Sixteen target mRNAs of these DKD-miRNAs were found to have a negative association with the estimated glomerular filtration rate (eGFR) in patients with DKD. The renal gene expression and eGFR data of DKD patients (n = 10-18) in the NephroSeq database were used. Based on these findings, 11 miRNA-mRNA regulatory networks were constructed for human DKD pathogenesis. Of these, in-vitro validation of miR-192-5p- CDKN3 (Cell cycle-dependent kinase inhibitor 3) network was done as miR-192-5p exhibited a maximum number of target genes in the identified DKD regulatory networks, and CDKN3 appeared as a novel target of miR-192-5p in our study. We demonstrated that miR-192-5p overexpression or knockdown of CDKN3 attenuated high glucose-induced apoptosis, fibrotic gene expression, cell hypertrophy, and cell cycle dysregulation and improved viability of proximal tubular cells. Moreover, miR-192-5p overexpression significantly inhibited CDKN3 mRNA and protein expression in proximal tubular cells. Overall, 11 miRNA-mRNA regulatory networks were predicted for human DKD pathogenesis; among these, the association of miR-192-5p- CDKN3 network DKD pathogenesis was confirmed in proximal tubular cell culture.
糖尿病肾病（DKD），又称糖尿病性肾病，是终末期肾病的主要病因。然而，在相当一部分2型糖尿病患者中，肾脏受累也可由非糖尿病原因引起（非糖尿病肾病，NDKD）。在本研究中，我们确定了人类DKD发病机制特有的miRNA-mRNA调控网络。对病例（DKD，n = 5）、疾病对照（伴有NDKD的T2DM，n = 6）和非糖尿病、非CKD对照（因肾癌接受肾切除术的患者，n = 3）的肾活检进行miRNA分析，发现了68种DKD特异性miRNA调控。发现这些DKD-miRNA的16个靶mRNA与DKD患者的估计肾小球滤过率（eGFR）呈负相关。使用了NephroSeq数据库中DKD患者（n = 10 - 18）的肾脏基因表达和eGFR数据。基于这些发现，构建了11个针对人类DKD发病机制的miRNA-mRNA调控网络。其中，对miR-192-5p - CDKN3（细胞周期依赖性激酶抑制剂3）网络进行了体外验证，因为miR-192-5p在已确定的DKD调控网络中表现出最多的靶基因，并且在我们的研究中CDKN3是miR-192-5p的一个新靶标。我们证明，miR-192-5p过表达或敲低CDKN3可减轻高糖诱导的细胞凋亡、纤维化基因表达、细胞肥大和细胞周期失调，并提高近端肾小管细胞的活力。此外，miR-192-5p过表达显著抑制近端肾小管细胞中CDKN3的mRNA和蛋白表达。总体而言，预测了11个针对人类DKD发病机制的miRNA-mRNA调控网络；其中，miR-192-5p - CDKN3网络与DKD发病机制的关联在近端肾小管细胞培养中得到了证实。
