The RPTEC/TERT1 Cell Line as an Improved Tool for In Vitro Nephrotoxicity Assessments

Simon-Friedt, Bridget R.; Wilson, Mark; Blake, Diane A.; Yu, Haixia; Eriksson, Yasmin; Wickliffe, Jeffrey K.

doi:10.1007/s12011-015-0339-y

Cited by 24 publications

(21 citation statements)

References 40 publications

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“…normal, healthy kidney, including significant gene expression changes and induced BPDE-DNA adducts (Simon-Friedt et al, 2015). Moreover, we were able to detect dA-AAI adduct levels in RPTEC/TERT1 cells over the whole AAI-concentration range.…”

Section: Discussionmentioning

confidence: 73%

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Comparison of Aristolochic acid I derived DNA adduct levels in human renal toxicity models

et al. 2019

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Aristolochic acid (AA) dependent human nephropathy results either from environmental exposure to Aristolochiaceae plant subspecies or their use in traditional phytotherapy. The toxic components are structurally related nitrophenanthrene carboxylic acids, i.e. Aristolochic acid I (AAI) and II (AAII). AAI is considered to be the major cause of Aristolochic acid nephropathy, characterized by severe renal fibrosis and upper urothelial cancer. Following enzymatic activation in kidney and/or liver, AAI metabolites react with genomic DNA to form persistent DNA adducts with purines. To determine whether AAI can be activated in human renal cells to form DNA adducts, we exposed telomerase immortalized renal proximal tubular epithelial cells (RPTEC/TERT1), the human embryonic kidney (HEK293) cell line, as well as primary human kidney cells (pHKC) to AAI in vitro. We modified an isotope dilution ultra-performance liquid chromatography/tandem mass spectrometry (ID-UPLC-MS/MS) based method for the quantification of dA-AAI adducts in genomic DNA. In addition, time dependent accumulation of adducts in renal cortex and bladder tissue from AAI/II treated Eker rats were used to validate the detection method. AAI-induced toxicity in human renal cells was determined by dA-AAI adduct quantification, the impact on cell viability, and NQO1 expression and activity. Our findings demonstrated adduct formation in all cell lines, although only pHKC and RPTEC/TERT1 expressed NQO1. The highest adduct formation was detected in pHKC despite low NQO1 expression, while we observed much lower adduct levels in NQO1-negative HEK293 cells. Adduct formation and decreased cell viability correlated only weakly. Therefore, our data suggested that i.) enzymes other than NQO1 could be at least equally important for AA bioactivation in human renal proximal tubule cells, and ii.) the suggested correlation between adduct levels and viability appears to be questionable.

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Section: Discussionmentioning

confidence: 73%

“…Substantial differences in activity of the involved DNA-repair pathway, i.e. nucleotide excision repair, can be ruled out as both RPTEC/TERT1 (Simon-Friedt et al, 2015) as well as HEK293 (Atanassov et al, 2004, Toga et al, 2014 are NER competent.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Aristolochic acid I derived DNA adduct levels in human renal toxicity models

et al. 2019

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“…Two important immortalized cell lines have been extensively used for nephrotoxicity screening: RPTEC/TERT1 and ciPTEC. The RPTEC/TERT1 cell line was immortalized using the human telomerase reverse transcriptase (hTERT) (Simon-Friedt et al 2015), whereas ciPTEC lines obtained either from urine or kidney tissue were transfected using hTERT and a temperature-sensitive mutant of SV large T antigen (SV40T), allowing these cells to proliferate at 33 °C and mature at 37 °C (Jansen et al 2014;Wilmer et al 2010) (Table 3). These cell lines surpass HK-2 cells by expressing all relevant markers cited above; however, OAT function was only attained after lentiviral transduction (Nieskens et al 2016).…”

Section: Conventional In Vitro Modelsmentioning

confidence: 99%

Kidney-based in vitro models for drug-induced toxicity testing

et al. 2019

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The kidney is frequently involved in adverse effects caused by exposure to foreign compounds, including drugs. An early prediction of those effects is crucial for allowing novel, safe drugs entering the market. Yet, in current pharmacotherapy, drug-induced nephrotoxicity accounts for up to 25% of the reported serious adverse effects, of which one-third is attributed to antimicrobials use. Adverse drug effects can be due to direct toxicity, for instance as a result of kidney-specific determinants, or indirectly by, e.g., vascular effects or crystals deposition. Currently used in vitro assays do not adequately predict in vivo observed effects, predominantly due to an inadequate preservation of the organs' microenvironment in the models applied. The kidney is highly complex, composed of a filter unit and a tubular segment, together containing over 20 different cell types. The tubular epithelium is highly polarized, and the maintenance of this polarity is critical for optimal functioning and response to environmental signals. Cell polarity is dependent on communication between cells, which includes paracrine and autocrine signals, as well as biomechanic and chemotactic processes. These processes all influence kidney cell proliferation, migration, and differentiation. For drug disposition studies, this microenvironment is essential for prediction of toxic responses. This review provides an overview of drug-induced injuries to the kidney, details on relevant and translational biomarkers, and advances in 3D cultures of human renal cells, including organoids and kidney-on-a-chip platforms.

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“…The lack of OAT function may be overcome by stably transfected lines available through American Type Culture Collection (ATCC, 2016), although currently limited data are available. RPTEC/TERT1 cells have been used for in vitro kidney toxicity assessments via genetic analysis (Aschauer et al, 2015b) and of environmental toxicants (Simon et al, 2014;Simon-Friedt et al, 2015). Recently, highly differentiated three-dimensional (3D) tubules of RPTEC/TERT1 cells were developed by sandwiching them between Matrigel layers, leading to a branched network of cell-free lumen; however, OAT1 expression was negligible, even though other characteristics of a well differentiated epithelium, such as polar expression of Na+/K+ ATPase and ZO-3, were present (Secker et al, 2017).…”

Section: Common Sources Of Kidney Cellsmentioning

confidence: 99%

Emerging Kidney Models to Investigate Metabolism, Transport, and Toxicity of Drugs and Xenobiotics

Bajaj¹,

Chowdhury²,

Yucha³

et al. 2018

Drug Metab Dispos

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The kidney is a major clearance organ of the body and is responsible for the elimination of many xenobiotics and prescription drugs. With its multitude of uptake and efflux transporters and metabolizing enzymes, the proximal tubule cell (PTC) in the nephron plays a key role in the disposition of xenobiotics and is also a primary site for toxicity. In this minireview, we first provide an overview of the major transporters and metabolizing enzymes in the PTCs responsible for biotransformation and disposition of drugs. Next, we discuss different cell sources that have been used to model PTCs in vitro, their pros and cons, and their characterization. As current technology is inadequate to evaluate reliably drug disposition and toxicity in the kidney, we then discuss recent advancements in kidney microphysiological systems (MPS) and the need to develop robust in vitro platforms that could be routinely used by pharmaceutical companies to screen compounds. Finally, we discuss the new and exciting field of stem cell-derived kidney models as potential cell sources for future kidney MPS. Given the push from both regulatory agencies and pharmaceutical companies to use more predictive "human-like" in vitro systems in the early stages of drug development to reduce attrition, these emerging models have the potential to be a game changer and may revolutionize how renal disposition and kidney toxicity in drug discovery are evaluated in the future.

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The RPTEC/TERT1 Cell Line as an Improved Tool for In Vitro Nephrotoxicity Assessments

Cited by 24 publications

References 40 publications

Comparison of Aristolochic acid I derived DNA adduct levels in human renal toxicity models

Comparison of Aristolochic acid I derived DNA adduct levels in human renal toxicity models

Kidney-based in vitro models for drug-induced toxicity testing

Emerging Kidney Models to Investigate Metabolism, Transport, and Toxicity of Drugs and Xenobiotics

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