Effect of gentamicin on the lysosomal system of cultured human proximal tubular cells

Regec, Annette L.; Trump, Benjamin F.; Trifillis, Anna L.

doi:10.1016/0006-2952(89)90098-1

Cited by 33 publications

(17 citation statements)

References 33 publications

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“…In Silico Prediction of the Extent of Lysosomal Trapping. For an accurate in silico prediction of lysosomal trapping, comprehensive information on the acidity and the fractional volume of the endo-/ lysosomal system is needed due to tremendous variations between cell types ranging from 0.23% up to 7.8% of the cellular volume (de Duve et al, 1974;Blouin et al, 1977;MacIntyre and Cutler, 1988a;Ufuk et al, 2015) with pH values reported in the range of 4.65-5.18 (Regec et al, 1989;Tietz et al, 1990;Kharbanda et al, 1997). We have, for the first time, determined a full profile of the pH values and corresponding fractional volumes of the endo-/lysosomal system in rat hepatocytes (Fig.…”

Section: Downloaded Frommentioning

confidence: 99%

Quantitation of Lysosomal Trapping of Basic Lipophilic Compounds Using In Vitro Assays and In Silico Predictions Based on the Determination of the Full pH Profile of the Endo-/Lysosomal System in Rat Hepatocytes

et al. 2018

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Lysosomal sequestration may affect the pharmacokinetics, efficacy, and safety of new basic lipophilic drug candidates potentially impacting their intracellular concentrations and tissue distribution. It may also be involved in drug-drug interactions, drug resistance, and phospholipidosis. However, currently there are no assays to evaluate the lysosomotropic behavior of compounds in a setting fully meeting the needs of drug discovery. We have, therefore, integrated a set of methods to reliably rank order, quantify, and calculate the extent of lysosomal sequestration in rat hepatocytes. An indirect fluorescence-based assay monitors the displacement of the fluorescence probe LysoTracker Red by test compounds. Using a lysosomal-specific evaluation algorithm allows one to generate IC 50 values at lower than previously reported concentrations. The concentration range directly agrees with the concentration dependency of the lysosomal drug content itself directly quantified by liquid chromatography-tandem mass spectrometry and thus permits a quantitative link between the indirect and the direct trapping assay. Furthermore, we have determined the full pH profile and corresponding volume fractions of the endo-/lysosomal system in plated rat hepatocytes, enabling a more accurate in silico prediction of the extent of lysosomal trapping based only on pK a values as input, allowing early predictions even prior to chemical synthesis. The concentration dependency-i.e., the saturability of the trappingcan then be determined by the IC 50 values generated in vitro. Thereby, a more quantitative assessment of the susceptibility of basic lipophilic compounds for lysosomal trapping is possible. This work was financed by Bayer AG, Germany. https://doi.org/10.1124/dmd.118.084541. s This article has supplemental material available at dmd.aspetjournals.org. ABBREVIATIONS: DAPI -49,6-diamidino-2-phenylindole; LC-MS/MS, liquid chromatography-tandem mass spectrometry; LTR, LysoTracker Red DND-99; PBS, phosphate-buffered saline. 49

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Section: Downloaded Frommentioning

confidence: 99%

Quantitation of Lysosomal Trapping of Basic Lipophilic Compounds Using In Vitro Assays and In Silico Predictions Based on the Determination of the Full pH Profile of the Endo-/Lysosomal System in Rat Hepatocytes

et al. 2018

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“…We can conclude that these effects are not important to toxicity in epithelial cells. When concentration of GM in endosomes reaches certain level, their membranes disrupts and their content including GM is released to cytoplasm (Regec et al, 1989[69]). In cytoplasm GM acts on mitochondria both directly and indirectly (Morales et al, 2010[53]) and activates intrinsic apoptosis pathway, breaks respiratory chain, decrease ATP synthesis and leads to oxidative stress by creating superoxide anion and hydroxyl radical (Cuzzocrea et al, 2002[15]) resulting in cell death.…”

Section: Tubular Effectsmentioning

confidence: 99%

Gentamicin nephrotoxicity in animals: current knowledge and future perspectives

Randjelović

Veljković

Stojiljković

et al. 2017

EXCLI Journal; 16:Doc388; ISSN 1611-2156

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“…Cytosolic gentamicin acts on mitochondria both directly and indirectly, activating the intrinsic pathway of apoptosis. 56 Other numerous disruptions take place, which further contributes to cell death. 48 …”

Section: Current Research Models Used To Study Akimentioning

confidence: 99%

New tides: using zebrafish to study renal regeneration

McCampbell

Wingert

2014

Translational Research

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Over the past several decades, the zebrafish has become one of the major vertebrate model organisms used in biomedical research. In this arena, the zebrafish has emerged as an applicable system for the study of kidney diseases and renal regeneration. The relevance of the zebrafish model for nephrology research has been increasingly appreciated as the understanding of zebrafish kidney structure, ontogeny, and the response to damage has steadily expanded. Recent studies have documented the amazing regenerative characteristics of the zebrafish kidney, which include the ability to replace epithelial populations after acute injury and to grow new renal functional units, termed nephrons. Here we discuss how nephron composition is conserved between zebrafish and mammals, and highlight how recent findings from zebrafish studies utilizing transgenic technologies and chemical genetics can complement traditional murine approaches in the effort to dissect how the kidney responds to acute damage and identify therapeutics that enhance human renal regeneration.

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Effect of gentamicin on the lysosomal system of cultured human proximal tubular cells

Cited by 33 publications

References 33 publications

Quantitation of Lysosomal Trapping of Basic Lipophilic Compounds Using In Vitro Assays and In Silico Predictions Based on the Determination of the Full pH Profile of the Endo-/Lysosomal System in Rat Hepatocytes

Quantitation of Lysosomal Trapping of Basic Lipophilic Compounds Using In Vitro Assays and In Silico Predictions Based on the Determination of the Full pH Profile of the Endo-/Lysosomal System in Rat Hepatocytes

Gentamicin nephrotoxicity in animals: current knowledge and future perspectives

New tides: using zebrafish to study renal regeneration

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