Nifurpirinol: A more potent and reliable substrate compared to metronidazole for nitroreductase‐mediated cell ablations

Bergemann, David; Massoz, Laura; Bourdouxhe, Jordane; Pardo, Claudio Andrès Carril; Voz, Marianne L.; Peers, Bernard; Manfroid, Isabelle

doi:10.1111/wrr.12633

Cited by 40 publications

(54 citation statements)

References 20 publications

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“…Of note, when higher dose assays were attempted, we observed near complete lethality with 24 h exposure at ≥5 μM NFP. This contrasts with a prior study reporting that lethality initiated at 15 μM NFP for 24 h treatments 32 .…”

Section: Ntr 20 Does Not Improve Nifurpirinol Ablation Efficacycontrasting

confidence: 99%

“…Contrary to the ~100-fold improvement observed with MTZ, no appreciable enhancement of cell ablation efficacy was observed with nifurpirinol ( Supplementary Fig. S5); cell loss being evident only at 2.5 μM NFP, akin to results with NTR 1.1 32 . This suggests that the enhanced prodrug activity of NTR 2.0 is specific to 5-nitroimidazole prodrugs (e.g.…”

Section: Ntr 20 Does Not Improve Nifurpirinol Ablation Efficacymentioning

confidence: 77%

Section: Discussionmentioning

confidence: 99%

“…Recently nifurpirinol (NFP) has been reported as an alternative prodrug that exhibits less variability in NTR-mediated cell ablation efficacy and is active at micromolar concentrations 32 . However, toxic effects were observed at 15 μM (24 h) and 10 µM (48 h) treatments 32 . In our hands, toxic effects arose at 5 μM NFP for 24 h. We do not know the reason for this discrepancy, but differing background strains or NFP source variation are possibilities.…”

Section: Discussionmentioning

confidence: 99%

“…In addition to MTZ, NTR enzymes can convert other prodrug substrates. For instance, a recent study demonstrated robust NTR 1.1-mediated cell ablation following a 24 h exposure to 2.5 μM nifurpirinol (NFP) 32 . To determine if NTR 2.0 exhibits improved NFP conversion activity, double transgenic larvae co-expressing YFP and NTR 2.0 (NRSE:KalTA4;UAS:YFP-NTR2.0) were exposed to a 1:10 dilution series of nifurpirinol (2.5 µM -2.5 nM) from 5 to 6 dpf, and YFP quantified at 7 dpf.…”

Section: Ntr 20 Does Not Improve Nifurpirinol Ablation Efficacymentioning

confidence: 99%

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NTR 2.0: a rationally-engineered prodrug converting enzyme with substantially enhanced efficacy for targeted cell ablation

Sharrock

Mulligan

Hall

et al. 2020

Preprint

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Heterologously-expressed bacterial nitroreductase (NTR) enzymes sensitize eukaryotic cells to prodrugs such as metronidazole (MTZ), enabling selective cell ablation paradigms that have expanded studies of cell function and regeneration in vertebrate systems. However, first-generation NTRs require confoundingly toxic prodrug treatments (e.g. 10 mM MTZ) and some cell types have proven resistant. We used rational engineering and cross-species screening to develop a NTR variant, NTR 2.0, which exhibits ~100-fold improvement in MTZ-mediated cell-specific ablation efficacy. Toxicity tests in zebrafish showed no deleterious effects of prolonged MTZ treatments of ≤1 mM. NTR 2.0 therefore enables sustained cell loss paradigms and ablation of previously resistant cell types. These properties permit enhanced interrogations of cell function, extended challenges to the regenerative capacities of discrete stem cell niches, and enable modeling of chronic degenerative diseases. Accordingly, we have created a series of bipartite transgenic resources to facilitate dissemination of NTR 2.0 to the research community.

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Section: Ntr 20 Does Not Improve Nifurpirinol Ablation Efficacycontrasting

confidence: 99%

Section: Ntr 20 Does Not Improve Nifurpirinol Ablation Efficacymentioning

confidence: 77%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Ntr 20 Does Not Improve Nifurpirinol Ablation Efficacymentioning

confidence: 99%

See 3 more Smart Citations

NTR 2.0: a rationally-engineered prodrug converting enzyme with substantially enhanced efficacy for targeted cell ablation

Sharrock

Mulligan

Hall

et al. 2020

Preprint

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Microscopic modulation and analysis of islets of Langerhans in living zebrafish larvae

et al. 2022

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Microscopic analysis of molecules and physiology in living cells and systems is a powerful tool in life sciences. While in vivo subcellular microscopic analysis of healthy and diseased human organs remains impossible, zebrafish larvae allow studying pathophysiology of many organs using in vivo microscopy. Here, we review the potential of the larval zebrafish pancreas in the context of islets of Langerhans and Type 1 diabetes. We highlight the match of zebrafish larvae with the expanding toolbox of fluorescent probes that monitor cell identity, fate and/or physiology in real time. Moreover, fast and efficient modulation and localization of fluorescence at a subcellular level, through fluorescence microscopy, including confocal and light sheet (single plane illumination) microscopes tailored to in vivo larval research, is addressed. These developments make the zebrafish larvae an extremely powerful research tool for translational research. We foresee that living larval zebrafish models will replace many cell line‐based studies in understanding the contribution of molecules, organelles and cells to organ pathophysiology in whole organisms.

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Microbial nitroreductases: A versatile tool for biomedical and environmental applications

Boddu

Perumal

Divakar³

2020

Biotechnology and Applied Biochemistry

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Nitroreductases, enzymes found mostly in bacteria and also in few eukaryotes, use nicotinamide adenine dinucleotide (NADH) or nicotinamide adenine dinucleotide phosphate (NADPH) as a cofactor for their activity and metabolize an enormous list of a diverse nitro group-containing compounds. Nitroreductases that are capable of metabolizing nitroaromatic and nitro heterocyclic compounds have drawn great attention in recent years owing to their biotechnological, biomedical, environmental, and human impact. These enzymes attracted medicinal chemists and pharmacologists because of their prodrug selectivity for activation/reduction of nitro compounds that wipe out pathogens/cancer cells, leaving the host/normal cells unharmed. It is applied in diverse fields of study like prodrug activation in treating cancer and leishmaniasis, designing fluorescent probes for hypoxia detection, cell imaging, ablation of specific cell types, biodegradation of nitro-pollutants, and interpretation of mutagenicity of nitro compounds. Keeping in view the immense prospects of these enzymes and a large number of research contributions in this area, the present review encompasses the enzymatic reaction mechanism, their role in antibiotic resistance, hypoxia sensing, cell imaging, cancer therapy, reduction of recalcitrant nitro chemicals, enzyme variants, and their specificity to substrates, reaction products, and their applications.

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Nifurpirinol: A more potent and reliable substrate compared to metronidazole for nitroreductase‐mediated cell ablations

Cited by 40 publications

References 20 publications

NTR 2.0: a rationally-engineered prodrug converting enzyme with substantially enhanced efficacy for targeted cell ablation

NTR 2.0: a rationally-engineered prodrug converting enzyme with substantially enhanced efficacy for targeted cell ablation

Microscopic modulation and analysis of islets of Langerhans in living zebrafish larvae

Microbial nitroreductases: A versatile tool for biomedical and environmental applications

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