Human MiR-4660 regulates the expression of alanine–glyoxylate aminotransferase and may be a biomarker for idiopathic oxalosis
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Dysfunction of oxalate synthesis can cause calcium oxalate stone disease and inherited primary hyperoxaluria (PH) disorders. PH type I (PH1) is one of the most severe hyperoxaluria disorders, which results in urolithiasis, nephrocalcinosis, and end-stage renal disease. Here, we sought to determine the role of microRNAs in regulating AGXT to contribute to the pathogenesis of mutation-negative idiopathic oxalosis.
We conducted bioinformatics to search for microRNAs binding to AGXT, and examined the expression of the highest hit (miR-4660) in serum samples of patients with oxalosis, liver tissue samples, and determined the correlation and regulation between the microRNA and AGXT in vitro.
MiR-4660 expression was downregulated in patients with oxalosis compared with healthy controls (84.03 copies/µL vs 33.02 copies/µL, P < 0.0001). Moreover, miR-4660 epigenetically decreased the expression of AGT in human liver tissues (Rho = − 0543, P = 0.037). Overexpression of miR-4660 in HepG2 and L02 cell lines led to dysregulation of AGXT at both the mRNA (by 71% and 81%, respectively; P < 0.001) and protein (by 49% and 42%, respectively; P < 0.0001) levels. We confirmed the direct target site of miR-4660 binding to the 3′UTR of AGXT by a luciferase assay.
MiR-4660 is probably a new biomarker for mutation-negative idiopathic oxalosis by regulating the post-transcription of AGXT, providing a potential treatment target of mutation-negative idiopathic oxalosis.
KeywordsIdiopathic oxalosis Primary hyperoxaluria MicroRNA Epigenetic regulation AGXT
This work was supported by grants from the National Natural Science Foundation of China (Nos. 91439109, 81700300, 81870176 and 91439109) and the Program for New Century Excellent Talents at the University of China (NCET-11-0181).
XT and DD designed the study. YZ, QL, XY and SS performed the experiments and statistical analysis. YY, ZD, YM, ZZ, XW and JJ collected the samples and took part in finishing the experiments.
Compliance with ethical standards
Conflict of interest
All authors do not have any actual or potential conflict of interest including any financial, personal or other relationships with other people or organizations within 3 years.
Human and animal rights
This study was approved by the appropriate local institutional review boards on human subject research at the Tongji Hospital of Huazhong University of Science and Technology (IRB ID: TJ-C20160114) and conformed to the guidelines set forth by the Declaration of Helsinki.
Written informed consent was obtained from all participants.
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