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Evaluating In Vitro Canine Kidney Slices as a Renal Toxicity Model Using Nephrotoxic Agents Cisplatin and Cadmium Chloride
Author(s) -
Kowalkowski Kenneth,
Klapczynski Marcin,
Blomme Eric,
Buck Wayne,
Liguori Michael
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.819.8
Subject(s) - toxicity , cadmium chloride , nephrotoxicity , chemistry , kidney , in vivo , viability assay , intracellular , pharmacology , necrosis , cisplatin , in vitro , pathology , cadmium , biochemistry , biology , medicine , endocrinology , microbiology and biotechnology , organic chemistry , chemotherapy
Precision‐cut tissue slices retain the multicellular, structural, and functional features of their original organ and may potentially be a more relevant approach to interrogate toxicity compared to traditional cell‐based in vitro systems. Before testing newly developed compounds, we investigated the ability of this system to detect kidney toxicity using prototypical kidney tubular toxicants cadmium chloride (CdCl 2 ) and cisplatin using a variety of endpoints. Kidneys from male beagle dogs were cut into approximately 300 mm slices. Primary experiments optimized the slicing conditions using ice cold Krebs‐Henseleit buffer with O 2 bubbled through the chamber to maintain viability. Slices were cultured in 95% O 2 /5% CO 2 atmosphere at 37°C for 24 to 48 hours. The slices were treated with vehicle or multiple doses of CdCl 2 and cisplatin. Historical toxicity endpoints evaluated were H&E stained sections, intracellular ATP content, LDH release, and RNA integrity. In vivo studies have proven kidney injury molecule‐1 (Kim‐1) is a valuable tool for detecting kidney toxicity, so mRNA levels via in situ hybridization were examined as a potential novel endpoint. LDH leakage was significantly elevated at all doses tested for both compounds. The intracellular levels of ATP also declined significantly at 100 mM CdCl 2 and 100mM cisplatin, indicative of substantial cellular damage. Mild to marked degeneration and necrosis of renal tubules was evident beginning at 24 h upon microscopic examination. Kim‐1, which is not expressed in normal tissue, was clearly overexpressed in CdCl 2 treated slices, especially along the tubular epithelium, but not in vehicle treated slices. Kim‐1 was undetectable in severely necrotic slices, likely due to RNA destabilization, later confirmed using integrity assessments. These data demonstrate that a renal slice in vitro model can be used to detect potent nephrotoxicants and that Kim‐1 should be further explored as a novel in vitro biomarker to monitor toxicity in renal slice systems. Disclosures: All authors are employees of AbbVie. The design, study conduct, and financial support for this research was provided by AbbVie. AbbVie participated in the interpretation of data, review, and approval of the publication.