A kidney injury molecule‐1 (Kim‐1) gene reporter in a mouse artificial chromosome: the responsiveness to cisplatin toxicity in immortalized mouse kidney S3 cells

Abstract

BackgroundKidney injury molecule‐1 (Kim‐1) has been validated as a urinary biomarker for acute and chronic renal damage. The expression of Kim‐1 mRNA is also activated by acute kidney injury induced by cisplatin in rodents and humans. To date, the measurement of Kim‐1 expression has not fully allowed the detection of in vitro cisplatin nephrotoxicity in immortalized culture cells, such as human kidney‐2 cells and immortalized proximal tubular epithelial cells.MethodsWe measured the augmentation of Kim‐1 mRNA expression after the addition of cisplatin using immortalized S3 cells established from the kidneys of transgenic mice harboring temperature‐sensitive large T antigen from Simian virus 40.ResultsA mouse Kim‐1 gene luciferase reporter in conjunction with an Hprt gene reporter detected cisplatin‐induced nephrotoxicity in S3 cells. These two reporter genes were contained in a mouse artificial chromosome, and two luciferases that emitted different wavelengths were used to monitor the respective gene expression. However, the Kim‐1 reporter gene failed to respond to cisplatin in A9 fibroblast cells that contained the same reporter mouse artificial chromosome, suggesting cell type‐specificity for activation of the reporter.ConclusionsWe report the feasibility of measuring in vitro cisplatin nephrotoxicity using a Kim‐1 reporter gene in S3 cells.

Fangjifuling Ameliorates Lipopolysaccharide-Induced Renal Injury via Inhibition of Inflammatory and Apoptotic Response in Mice

“…Two of the most promising biomarkers of AKI are KIM-1 [19] and NGAL [20]. We found that pretreatment with FF inhibited the overexpression of KIM-1 and NGAL in renal tissue of LPS-induced AKI mice.…”

“…We found that pretreatment with FF inhibited the overexpression of KIM-1 and NGAL in renal tissue of LPS-induced AKI mice. The release of KIM-1 and NGAL from damaged renal tissue also leads to the downstream activation of transcription factors that regulate the expression of inflammatory cytokines and chemokines [19,21].…”

Combinations of chromosome transfer and genome editing for the development of cell/animal models of human disease and humanized animal models

“…The MI-MAC has also been transferred into HepG2 cells to verify cytotoxicity levels via luciferase reporter assay [69]. The MI-HAC/MAC system can be used in various cell types and has successfully achieved the insertion of large GLVs such as PACs and BACs and multiple genes into the MI-HAC/ MAC [70][71][72]. Although limitations exist regarding the available drug-resistant genes for five GLVs loading onto the MI-HAC/MAC vector, this was solved by reusing the same drug-resistant genes by disrupting them with genome editing technology (Honma et al, 2017, unpublished data).…”

CRISPR/Cas9 engineering of a KIM-1 reporter human proximal tubule cell line

“…KIM-1 has been found to be a useful biomarker for kidney injury [ 3 , 4 ]. In vivo , KIM-1 can be upregulated in proximal tubular cells in response to a variety of injuries [ 5 , 6 ]. A greater understanding of what regulates the KIM-1 locus could lead to more insight into kidney injury.…”