Quantification of Natural Growth of Two Strains of Mycobacterium Marinum for Translational Antituberculosis Drug Development

Abstract

The zebrafish infected with Mycobacterium marinum (M. marinum) is an attractive tuberculosis disease model, showing similar pathogenesis to Mycobacterium tuberculosis (M. tuberculosis) infections in humans. To translate pharmacological findings from this disease model to higher vertebrates, a quantitative understanding of the natural growth of M. marinum in comparison to the natural growth of M. tuberculosis is essential. Here, the natural growth of two strains of M. marinum, E11 and M USA , is studied over an extended period using an established model-based approach, the multistate tuberculosis pharmacometric (MTP) model, for comparison to that of M. tuberculosis. Poikilotherm-derived strain E11 and human-derived strain M USA were grown undisturbed up to 221 days and viability of cultures (colony forming unit (CFU)/mL) was determined by plating at different time points. Nonlinear mixed effects modeling using the MTP model quantified the bacterial growth, the transfer among fast, slow, and non-multiplying states, and the inoculi. Both strains showed initial logistic growth, reaching a maximum after 20-25 days for E11 and M USA , respectively, followed by a decrease to a new plateau. Natural growth of both E11 and M USA was best described with Gompertz growth functions. For E11, the inoculum was best described in the slow-multiplying state, for M USA in the fast-multiplying state. Natural growth of E11 was most similar to that of M. tuberculosis, whereas M USA showed more aggressive growth behavior. Characterization of natural growth of M. marinum and quantitative comparison with M. tuberculosis brings the zebrafish tuberculosis disease model closer to the quantitative translational pipeline of antituberculosis drug development. The zebrafish (Danio rerio) is an increasingly utilized disease model organism to study tuberculosis. 1,2 Infections of zebrafish embryos with Mycobacterium marinum (M. marinum), a close relative of Mycobacterium tuberculosis (M. tuberculosis), 3 show similar pathogenesis to M. tuberculosis infection in humans. 4 The zebrafish as a disease model organism has several

Anti‐tuberculosis effect of isoniazid scales accurately from zebrafish to humans

“…The drug responses of isoniazid, rifampicin and ethambutol have been quantified on these states (Clewe, Aulin, Hu, Coates, & Simonsson, 2016) and successfully translated to mice (Chen et al, 2017) and patients (Clewe et al, 2018;Svensson & Simonsson, 2016). This multistate approach, especially when the in vitro natural growth and time-kill is repeated for M. marinum as well, can be integrated with our analysis of M. marinum to strengthen the translational value of findings in zebrafish to higher vertebrates (Van Wijk, Van der Sar, et al, 2020).…”

Standardization of zebrafish drug testing parameters for muscle diseases

“…Assaying internal concentrations can provide valuable information on calculating various pharmacokinetic parameters to further translate drug screening findings to humans ( Rennekamp and Peterson, 2015 ; Yadav et al, 2021 ). In particular, methods for nanoscale blood sampling in zebrafish larvae and pharmacokinetic-pharmacodynamic modeling demonstrated that the larval zebrafish response to an anti-tuberculosis drug could be translated to known effects in humans ( Van Wijk et al, 2020 ). Despite these advances, measuring the biodistribution of compounds to specific organ systems in zebrafish embryos and larvae is difficult.…”

Drug Resistance in Nontuberculous Mycobacteria: Mechanisms and Models

“…Similarly, two patients infected with the same strain can have different susceptibility to the same antibiotic because of differences in immunity [ 140 ]. It is therefore recommended to validate drug susceptibilities in a panel of NTM isolates to increase the chances of selecting strains with the closest growth profile as the isolate of interest [ 131 , 141 ].…”