Increased levels of urinary phenylacetylglycine associated with mitochondrial toxicity in a model of drug-induced phospholipidosis

Abstract

Background: Phospholipidosis (PLD) is a lysosomal storage disorder induced by a class of cationic amphiphilic drugs. However, drug-induced PLD is reversible. Evidence of PLD from animal studies with some compounds has led to discontinuation of development. Regulatory authorities are likely to request additional studies when PLD is linked to toxicity. Objective: We conducted a trial to investigate urinary phenylacetylglycine (uPAG) as a biomarker for PLD. Materials and methods: Five groups of 12 male Wistar rats were dosed once with vehicle, 300 mg/kg or 1500 mg/kg of compound A (known to induce PLD), or 300 mg/kg or 1000 mg/ kg of compound B (similar structure, but does not induce PLD) to achieve similar plasma exposures. Following dosing, urine and blood samples underwent nuclear magnetic resonance (NMR), proteomic, and biochemical analyses. Necropsies were performed at 48 and 168 h, organ histopathology evaluated, and gene expression in liver analyzed by microarray. Electron microscopic examination of peripheral lymphocytes was performed. Results: For compound A, uPAG increased with dose, correlating with lamellar inclusion bodies formation in peripheral lymphocytes. NMR analysis showed decreased tricarboxylic acid cycle intermediates, inferring mitochondrial toxicity. Mitochondrial dysfunction was suggested by uPAG increase, resulting from a switch to anaerobic metabolism or disruption of the urea cycle. Discussion and conclusion: uPAG shows utility as a noninvasive biomarker for mitochondrial toxicity associated with drug-induced PLD, providing a mechanistic hypothesis for toxicity associated with PLD likely resulting from combined direct and indirect mitochondrial toxicity via impairment of the proton motor force and alteration of fatty acid catabolism.

Effects of long-term cadmium exposure on urinary metabolite profiles in mice

“…Homoserine is an excreted metabolite produced in the liver from the abnormal methionine production (Gazarian et al, 2002). Increased levels of urinary phenylacetylglycine associated with drug-induced phospholipidosis (Doessegger et al, 2013). (S)-2-Acetolactate is an intermediate in the biosynthesis of valine, leucine, and isoleucine.…”

Nuclear magnetic resonance‐ and mass spectrometry‐based metabolomics to study maleic acid toxicity from repeated dose exposure in rats

“…N‐PAG is an acyl‐glycine classified as minor metabolites of fatty acids; statistically significant elevated levels were detected in the diurnal urine samples from the high‐dose group. Elevated levels of this metabolite in animals demonstrated abnormal phospholipid accumulation in the lungs, liver and kidneys, and are associated with mitochondrial toxicity and phospholipidosis (Delaney et al, ; Doessegger et al, ). The markedly enlarged and vacuolated renal cells from our histological analysis indicate MA may instigate mitochondrial toxicity and phospholipidosis (Reasor & Kacew, ).…”

Unravelling the metabolic impact of SBS-associated microbial dysbiosis: Insights from the piglet short bowel syndrome model

“…Increased acetyl carnitine levels are reported to differentiate steatosis from NASH 33 and correlate with the clinical stage of hepatocellular carcinoma 34 . Phenylacetylglycine is a reported biomarker of abnormal phospholipid accumulation (phopholipidosis) 35 36 suggesting the current results may reflect the extensive hepatic fat accumulation observed in the piglet SBS-ALD model 15 .…”