A multidimensional metabolomics workflow to image biodistribution and evaluate pharmacodynamics in adult zebrafish

Abstract

An integrated evaluation of the tissue distribution and pharmacodynamic properties of a therapeutic is essential for successful translation to the clinic. To date, however, cost-effective methods to measure these parameters at the systems level in model organisms are lacking. Here, we introduce a multidimensional workflow to evaluate drug activity that combines mass spectrometry-based imaging, absolute drug quantitation across different biological matrices, in vivo isotope tracing and global metabolome analysis in the adult zebrafish. As a proof of concept, we quantitatively determined the whole-body distribution of the anti-rheumatic agent hydroxychloroquine sulfate (HCQ) and measured the systemic metabolic impacts of drug treatment. We found that HCQ distributed to most organs in the adult zebrafish 24 h after addition of the drug to water, with the highest accumulation of both the drug and its metabolites being in the liver, intestine and kidney. Interestingly, HCQ treatment induced organ-specific alterations in metabolism. In the brain, for example, HCQ uniquely elevated pyruvate carboxylase activity to support increased synthesis of the neuronal metabolite, N-acetylaspartate. Taken together, this work validates a multidimensional metabolomics platform for evaluating the mode of action of a drug and its potential off-target effects in the adult zebrafish. This article has an associated First Person interview with the first author of the paper.

Using mass spectrometry imaging to map fluxes quantitatively in the tumor ecosystem

“…None of the mass spectrometry methods that we applied in this study enabled direct measurement of acetyl carbon labeling in acetyl-CoA. While cells labeled in culture with U- 13 C glucose mostly produce acetyl-CoA in which the acetyl group is M0 or M2, the M1 species tends to be more prevalent in vivo, possibly due to increased malic enzyme activity or recycling of endogenous CO 2 33 , 50 – 52 . Indirect evidence of M1 and M2 acetyl-CoA in our animals is provided by citrate and palmitate labeling as measured by LC/MS from bulk tissue (Fig.…”

Standardization of zebrafish drug testing parameters for muscle diseases

“…In published studies of zebrafish muscle disease drug screens, the concentrations of small molecules applied to embryos have typically ranged from 1 to 33 μM ( Tables 1 and 2 ). Concentrations greater than 50 μM can often elicit adverse events or off-target effects and are rarely used in high-throughput drug library screens in zebrafish ( Jackstadt et al, 2022 ; Wittmann et al, 2012 ; Tran et al, 2007 ). When wild-type zebrafish embryos were used to evaluate the toxicity of a chemical compound library with a concentration range of 100 nM to 10 μM, most chemicals showed little or no adverse effect on embryo health and survival at 1 μM ( Farr et al, 2020 ).…”

Adult zebrafish as advanced models of human disease

“…Drug testing in the adult zebrafish offer a new opportunity to understand how drugs function, and how they are metabolized within a whole animal and in specific organs and tissues. Gary Patti and Leah Shriver's laboratories have recently developed a multidimensional metabolomics platform to investigate the mode of action and toxicity of drugs in adult zebrafish by combining an array of techniques, including mass spectrometry and in vivo isotope tracing (Jackstadt et al, 2022). In this context, there are two exciting advantages of the adult zebrafish: individual adult zebrafish organs can be dissected and evaluated to directly assess drug biodistribution and pharmacodynamics.…”