Multiplexed assay panel of cytotoxicity in HK-2 cells for detection of renal proximal tubule injury potential of compounds

Yang, Wu; Connors, David; Barber, Lauren E.; Jayachandra, Sukhanya; Hanumegowda, Umesh; Adams, Stephen P.

doi:10.1016/j.tiv.2009.06.003

Cited by 72 publications

(67 citation statements)

References 45 publications

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“…Interestingly, HK-2 cells, an HPV-immortalized human renal proximal tubule cell line, were shown to have an IC 50 of 20 pg/ml at 72 h posttreatment with Stx2, compared to 2.2 pg/ml and 0.7 pg/ml at the same time point for 2D and 3D NKi-2 cells, respectively, in the present study. HK-2 cells have also been shown to lack sensitivity to drug-induced toxicity (26), indicating that they may not be the best cell line for studying renal cytotoxicity.…”

Section: Discussionmentioning

confidence: 99%

“…To overcome these limitations, investigators have used either animal models (18,19) or two-dimensional (2D) monolayer cell cultures to study Stx-mediated disease (20)(21)(22). 2D cell culture systems, while simple and low in cost, lack the architecture required to play a major role in cell-cell and cellextracellular matrix (ECM) interactions and are thus incapable of recapitulating the complexity of the in vivo environment (23,24), as well as inadequate at predicting cellular toxicity (25,26). Studies using 2D human renal proximal tubule cells have demonstrated that Stx increases apoptosis in a time-and dose-dependent manner, along with the secretion of IL-1␤, IL-6, and IL-8, the inhibition of protein synthesis, and water absorption (20,21,27,28).…”

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confidence: 99%

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Effects of Shiga Toxin Type 2 on a Bioengineered Three-Dimensional Model of Human Renal Tissue

et al. 2015

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dShiga toxins (Stx) are a family of cytotoxic proteins that can cause hemolytic-uremic syndrome (HUS), a thrombotic microangiopathy, following infections by Shiga toxin-producing Escherichia coli (STEC). Renal failure is a key feature of HUS and a major cause of childhood renal failure worldwide. There are currently no specific therapies for STEC-associated HUS, and the mechanism of Stx-induced renal injury is not well understood primarily due to a lack of fully representative animal models and an inability to monitor disease progression on a molecular or cellular level in humans at early stages. Three-dimensional (3D) tissue models have been shown to be more in vivo-like in their phenotype and physiology than 2D cultures for numerous disease models, including cancer and polycystic kidney disease. It is unknown whether exposure of a 3D renal tissue model to Stx will yield a more in vivo-like response than 2D cell culture. In this study, we characterized Stx2-mediated cytotoxicity in a bioengineered 3D human renal tissue model previously shown to be a predictor of drug-induced nephrotoxicity and compared its response to Stx2 exposure in 2D cell culture. Our results demonstrate that although many mechanistic aspects of cytotoxicity were similar between 3D and 2D, treatment of the 3D tissues with Stx resulted in an elevated secretion of the kidney injury marker 1 (Kim-1) and the cytokine interleukin-8 compared to the 2D cell cultures. This study represents the first application of 3D tissues for the study of Stx-mediated kidney injury.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Effects of Shiga Toxin Type 2 on a Bioengineered Three-Dimensional Model of Human Renal Tissue

et al. 2015

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“…Nanoparticles often become enriched in the kidney. Transformed or immortalized animal and human renal proximal tubule cells are frequently used for in vitro nephrotoxicology, and primary proximal tubule cells have also been applied (Chen et al 1990;Bach et al 1997;Pfaller and Gstraunthaler 1998;Li et al 2003Li et al , 2006McGoldrick et al 2003;Pallet et al 2008;Hovda et al 2009;Konigs et al 2009;Wu et al 2009). Herein we investigated the effects of pHF-QDs on well-established porcine and human proximal tubule cell lines, and on primary human proximal tubule cells obtained from different donors and vendors, in order to learn more about the advantages and disadvantages of each cell model.…”

Section: Discussionmentioning

confidence: 99%

“…Also, the use of permanent lines of human cells is not without problems. It has been found that cell lines show reduced sensitivity to toxins and toxic effects of nanoparticles, as compared to primary human cells (Bregoli et al 2009;Wu et al 2009), suggesting that the use of primary human cells might be more appropriate.…”

Section: Introductionmentioning

confidence: 99%

Effects of quantum dots on different renal proximal tubule cell models and on gel-free renal tubules generatedin vitro

Zheng

Zhang

et al. 2011

Nanotoxicology

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We investigated the interactions of different types of human and porcine renal proximal tubule-derived cells with core-shell CdSe@ZnS quantum dots (QDs) coated with polymerized histidine-formaldehyde (pHF). The results revealed that porcine and human proximal tubule cells showed a markedly different uptake behavior. This applied to flat epithelial monolayers, as well as to proximal tubules formed on two-dimensional (2D) surfaces in vitro. Primary human cells were most sensitive to the cytotoxic effects of QDs, but displayed inter-donor variability, which appeared to depend on the state of differentiation. The results suggested that human proximal tubule-derived cells were more appropriate than porcine cells for in vitro nanotoxicology. Primary human cells might be suitable when their state of differentiation and inter-donor variability were well-controlled. Furthermore, the results suggested that gel-free proximal tubules formed in vitro could be used as test system to address uptake and transport of nanometer-sized particles in human renal structures.

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“…Morelli et al [33] validated an HCA screen for phospholipidotic potential of cationic amphiphilic drugs. Application of a multiplex assay analogous to the quadprobe HCA assay with measurements for mitochondrial activity, apoptosis, plasma membrane permeability and DNA in human kidney cells (HK-2) revealed proximal tubule injury potential of compounds [34]. Several studies have demonstrated that neurotoxicity can be effectively screened for using HCA analysis of rat PC12, mouse N1E-115 and human SH-SY5Y neuronal cell lines for assessment of neurite outgrowth [35] and inhibition of cell proliferation [36].…”

Section: M72mentioning

confidence: 99%

High‐Content Analysis in Toxicology: Screening Substances for Human Toxicity Potential, Elucidating Subcellular Mechanisms and In Vivo Use as Translational Safety Biomarkers

O’Brien

2014

Basic Clin Pharma Tox

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High-content analysis (HCA) of in vitro biochemical and morphological effects of classic (small molecule) drugs and chemicals is concordant with potential for human toxicity. For hepatotoxicity, concordance is greater for cytotoxic effects assessed by HCA than for conventional cytotoxicity tests and for regulatory animal toxicity studies. Additionally, HCA identifies chronic toxicity potential, and drugs producing idiosyncratic adverse reactions and/or toxic metabolites are also identified by HCA. Mechanistic information on the subcellular basis for the toxicity is frequently identified, including various mitochondrial effects, oxidative stress, calcium dyshomeostasis, phospholipidosis, apoptosis and antiproliferative effects, and a fingerprinting of the sequence and pattern of subcellular events. As these effects are frequently non-specific and affect many cell types, some toxicities may be detected and monitored by HCA of peripheral blood cells, such as for anticancer and anti-infective drugs. Critical methodological and interpretive features are identified that are critical to the effectiveness of the HCA cytotoxicity assessment, including the need for multiple days of exposure of cells to drug, use of a human hepatocyte cell line with metabolic competence, assessment of multiple pre-lethal effects in individual live cells, consideration of hormesis, the need for interpretation of relevance of cytotoxicity concentration compared to efficacy concentration and quality management. Limitations of the HCA include assessment of drugs that act on receptors, transporters or processes not found in hepatocytes. HCA may be used in a) screening lead candidates for potential human toxicity in drug discovery alongside of in vitro assessment of efficacy and pharmacokinetics, b) elucidating mechanisms of toxicity and c) monitoring in vivo toxicity of drugs with known toxicity of known mechanism.Classic (small molecule) drug toxicity is one of the major challenges for the pharmaceutical industry, not only because of the associated loss of human life or health, but because of enormous financial loss of past investment and of future revenue potential. Attrition of drugs is progressively more costly the later it occurs. It costs almost two billion dollars to bring the average drug through discovery and development to registration [1]. Furthermore, typically 10-12 years of investment of time by hundreds of pharmaceutical staff will have been mis-spent.Drug safety has been an important cause of this attrition. Analysis of safety attrition of approved drugs over a 25-year period ending in 1999 indicated an annual average of 0.7 approved drugs was withdrawn and two drugs received black box warnings [2]. From 1994 to 2006, the average safety attrition was even higher with 2.1 drugs per year, with 38 drugs withdrawn from the market by the US Food and Drug Agency More than 80% of market withdrawals due to drug toxicity were due to cardiotoxicity and hepatotoxicity. In 2003, druginduced liver injury was found to be the most fr...

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Multiplexed assay panel of cytotoxicity in HK-2 cells for detection of renal proximal tubule injury potential of compounds

Cited by 72 publications

References 45 publications

Effects of Shiga Toxin Type 2 on a Bioengineered Three-Dimensional Model of Human Renal Tissue

Effects of Shiga Toxin Type 2 on a Bioengineered Three-Dimensional Model of Human Renal Tissue

Effects of quantum dots on different renal proximal tubule cell models and on gel-free renal tubules generatedin vitro

High‐Content Analysis in Toxicology: Screening Substances for Human Toxicity Potential, Elucidating Subcellular Mechanisms and In Vivo Use as Translational Safety Biomarkers

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