The miR-15a-5p-XIST-CUL3 regulatory axis is important for sepsis-induced acute kidney injury

Abstract

Background: Acute kidney injury (AKI) refers to a sudden loss of renal function. This study was performed to identify the key RNAs acting in the mechanism of sepsis-induced AKI.Methods: Microarray dataset GSE94717 (including six sepsis-induced AKI samples and three control samples) was downloaded from Gene Expression Omnibus database. Differentially expressed miRNAs (DE-miRNAs) were identified. The miRNA targets were predicted and enrichment analysis was performed. Protein-protein interaction (PPI) and competing endogenous RNA (ceRNA) regulatory networks were constructed. Mouse podocytes were treated with lipopolysaccharide (LPS), following by cell viability and PCR analysis. Cellular apoptosis and the ceRNA network were validated.Results: Thirty-one common DE-miRNAs (two up-regulated and 29 down-regulated) by AKI versus control and male AKI versus control were identified. We found the targets of miR-15a-5p, miR-15b-5p, and miR-16-5p were involved in mTOR signaling pathway, and those of miR-29b-3p and miR-16-5p were enriched in PI3K-Akt signaling pathway. RNAs including miR-15b-5p, miR-15a-5p, miR-107, XIST, miR-16-5p, and cullin 3 gene (CUL3) were included in the ceRNA regulatory network. The downregulation of miR-15a-5p and miR-15b-5p and the upregulation of lncRNA XIST and CUL3 gene were validated using qPCR. The miR-15a-5p-XIST-CUL3 regulatory axis was identified and was validated. We confirmed that LPS inhibited the growth of mouse podocytes and seven of the ten miRNAs, but upregulated XIST and CUL3. Transfection analysis showed XIST siRNA enhanced LPS-induced MPC5 cell apoptosis and miR-15a-5p inhibitor reserved it, so did as CUL3 overexpression for miR-15a-5p mimics.Conclusion: The miR-15a-5p-XIST-CUL3 regulatory axis was related to the pathogenesis of sepsis-induced AKI.HighlightsTotally, 31 miRNAs were dysregulated between disease and control groups.MiR-15a-5p, miR-15b-5p, and miR-16-5p were involved in mTOR signaling pathway.MiR-16-5p and miR-29b-3p were implicated in PI3K-Akt signaling pathway.The miR-15a-5p-XIST-CUL3 axis was critical for sepsis-induced AKI.

Downregulation of XIST ameliorates acute kidney injury by sponging miR‐142‐5p and targeting PDCD4

“…Cheng & Wang, 2020). The XIST/miR-15a-5p/CUL3 regulatory axis participates in sepsis-induced AKI (Xu et al, 2019). Here, we found that XIST was upregulated in AKI and knockdown of XIST Recently, accumulating evidence has shown that miR-142-5p is involved in various diseases (Chaudhuri, Yelamanchili, Marcondes, & Fox, 2013;Schaefer, Montufar-Solis, Vigneswaran, & Klein, 2011;Zhu et al, 2016).…”

Detection of urinary miRNAs for diagnosis of clear cell renal cell carcinoma

“…Indeed, when we tested the presence of RNU6-2 in the urine, about half of the urine samples from HSs did not show a Ct < 35 (not shown) and we believe it is risky to normalize miRNAs with a transcript present in the urine at very low levels and with high inter-subject variability, particularly in HSs. Moreover, miR-15a is present at high levels in the blood of patients with acute kidney injury 39 , suggesting that miR-15a may be present at high levels in the urine of patients without cancer but with non-cancerous diseases of the kidney, potentially increasing the false positive rate of the test. Our study suggests that evaluating the amplification of some miRNAs instead of that of one miRNA represents an advantage in patients in which alteration of the urinary presence of just one miRNA occurs for reasons that are not evident in clinics.…”

Acute Kidney Injury in Septic Patients Treated by Selected Nephrotoxic Antibiotic Agents—Pathophysiology and Biomarkers—A Review

“…The inhibition of XIST and miR-15a-5p enhanced and preserved LPS-induced apoptosis significantly, while the miR-15a-5p inhibitor reversed the renal cell apoptosis. Furthermore, overexpression of the CUL3 gene considerably reduced the LPS and miR-l5a-5p -induced apoptosis [ 57 ]. Another explanation of sepsis pathophysiology is based on regulation of the crucial inflammatory response of damaged organs with the participation of miR-15a-5p .…”