tRNA-Derived Fragments in Podocytes with Adriamycin-Induced Injury Reveal the Potential Mechanism of Idiopathic Nephrotic Syndrome

Abstract

Idiopathic nephrotic syndrome (INS) is a disease involving injury to podocytes in the glomerular filtration barrier, and its specific causes have not been elucidated. Transfer RNA-derived fragments (tRFs), products of precise tRNA cleavage, have been indicated to play critical roles in various diseases. Currently, there is no relevant research on the role of tRFs in INS. This study intends to explore the changes in and importance of tRFs during podocyte injury in vitro and to further analyze the potential mechanism of INS. Differentially expressed tRFs in the adriamycin-treated group were identified by high-throughput sequencing and further verified by quantitative RT-PCR. In total, 203 tRFs with significant differential expression were identified, namely, 102 upregulated tRFs and 101 downregulated tRFs (q<0.05, ∣log2FC∣≥2). In particular, AS-tDR-008924, AS-tDR-011690, tDR-003634, AS-tDR-013354, tDR-011031, AS-tDR-001008, and AS-tDR-007319 were predicted to be involved in podocyte injury by targeting the Gpr, Wnt, Rac1, and other genes. Furthermore, gene ontology analysis showed that these differential tRFs were strongly associated with podocyte injury processes such as protein binding, cell adhesion, synapses, the actin cytoskeleton, and insulin-activate receptor activity. KEGG pathway analysis predicted that they participated in the PI3K-Akt signaling pathway, Wnt signaling pathway, and Ras signaling pathway. It was reported that these pathways contribute to podocyte injury. In conclusion, our study revealed that changes in the expression levels of tRFs might be involved in INS. Seven of the differentially expressed tRFs might play important roles in the process of podocyte injury and are worthy of further study.

Expression profiles of tRNA‑derived fragments in high glucose‑treated tubular epithelial cells

“…revealed the potential mechanism of idiopathic nephrotic syndrome (12,13). To the best of our knowledge, however, little is known about the role of tRFs in TIF.…”

“…Additionally, tRFs may participate in the early diagnosis of cancer by serving as prognostic biomarkers (10). A growing number of studies have demonstrated that tRFs may be associated with chronic kidney disease (CKD) progression (11)(12)(13). Khurana et al (11) identified that tRF Val and tRF Leu , derived from urinary exosomes, are suitable biomarkers for the early diagnosis of CKD.…”

Dysregulation of tRNA-derived small RNAs and their potential roles in lupus nephritis

“…Nowadays, increasing studies have identified and emphasized the important role of tsRNAs in various diseases. 7,9,16,17 Therefore, we herein aimed to explore the expression pattern of tsRNAs, as well as the potential roles of candidate tsRNAs in the development of LN. Of note, our sequencing and qRT-PCR data showed that the expression levels and subtypes of tsRNAs were significantly altered in renal tissues of LN patients compared with the control.…”

“…7,8 Interestingly, mounting evidence has indicated that tsRNAs are deeply implicated in human diseases, such as cancer, autoimmune diseases, and neurological diseases, rather than the byproducts of random tRNA cleavage. [9][10][11][12] Nevertheless, to date, no studies are focusing on the changes of tsRNAs expression profile in local renal tissues in LN, as well as their potential functional roles in the pathogenesis of LN. In this study, we first identified the expression profiles of tsRNAs in kidney tissues from the LN and control groups through high-throughput sequencing. We subsequently validated candidate tsRNAs using quantitative real-time PCR (qRT-PCR).…”

miR‑199b‑5p mediates adriamycin‑induced podocyte apoptosis by inhibiting the expression of RGS10

“…The miR-199b-5p mimic (sense, 5'-CCCAGU GUUUAGACUACCUGUUC-3'; antisense, 5'-ACAGGUAGU CUAAACACUGGGUU-3'; GenePharma, Shanghai, China), miR-199b-5p inhibitor (5'-GAACAGGUAGUCUAAACACU GGG-3'; Shanghai GenePharma Co., Ltd.), negative control mimic (sense, 5'-UUCUCCGAACGUGUCACGUTT-3'; antisense, 5'-ACGUGACACGUUCGGAGAATT-3'; Shanghai GenePharma Co., Ltd.), and negative control inhibitor (5'-CAG UACUUUUGUGUAGUACAA-3'; Shanghai GenePharma Co., Ltd.) were transfected for 6 h at a concentration of 100 nM using Lipofectamine 2000 transfection reagent (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. The medium was changed again, and 1 µg/ml ADR (MedChemExpress) was added for 24 h treatment ( 21 ). To verify the effect of AKT/mTOR pathway on ADR-induced podocyte apoptosis, the differentiated podocytes were treated with 1 mg/l rapamycin (RAPA; MedChemExpress) after ADR intervention.…”