A New Transgenic Line for Rapid and Complete Neutrophil Ablation

Overview

Journal Zebrafish

Date 2022 May 26

PMID 35617702

Authors

Christopher J Hall

Jonathan W Astin

Jeff S Mumm

David F Ackerley

Affiliations

Soon will be listed here.

Abstract

Zebrafish lines expressing nitroreductase (NTR) in specific cell compartments, which sensitizes those cells to metronidazole (MTZ)-mediated ablation, have proven extremely useful for studying tissue regeneration and investigating cell function. In contrast to many cells, neutrophils are comparatively resistant to the NTR/MTZ targeted ablation strategy. Recently, a rationally engineered variant of NTR (NTR 2.0) has been described that exhibits greatly improved MTZ-mediated ablation efficacy in zebrafish. We show that a transgenic line with neutrophil-restricted expression of NTR 2.0 demonstrates complete neutrophil ablation, with an MTZ dose 100-fold less than current treatment regimens, and with treatment durations as short as 5 h.

Citing Articles

Neutrophils facilitate the epicardial regenerative response after zebrafish heart injury.

Peterson E, Sun J, Chen X, Wang J Dev Biol. 2024; 508:93-106.

PMID: 38286185 PMC: 10923159. DOI: 10.1016/j.ydbio.2024.01.011.

Infection-experienced HSPCs protect against infections by generating neutrophils with enhanced mitochondrial bactericidal activity.

Darroch H, Keerthisinghe P, Sung Y, Rolland L, Prankerd-Gough A, Crosier P Sci Adv. 2023; 9(36):eadf9904.

PMID: 37672586 PMC: 10482338. DOI: 10.1126/sciadv.adf9904.

An inducible model of chronic hyperglycemia.

Tucker T, Knitter C, Khoury D, Eshghi S, Tran S, Sharrock A Dis Model Mech. 2023; 16(8).

PMID: 37401381 PMC: 10417516. DOI: 10.1242/dmm.050215.

gene promoter-driven ciliopathy disease model in zebrafish.

Lee M, Han H, Choi T, Lee K, Baasankhuu A, Kim H Front Cell Dev Biol. 2023; 11:1200599.

PMID: 37363725 PMC: 10285392. DOI: 10.3389/fcell.2023.1200599.

References

Sharrock A, Mulligan T, Hall K, Williams E, White D, Zhang L . NTR 2.0: a rationally engineered prodrug-converting enzyme with substantially enhanced efficacy for targeted cell ablation. Nat Methods. 2022; 19(2):205-215. PMC: 8851868. DOI: 10.1038/s41592-021-01364-4. View

Renshaw S, Loynes C, Trushell D, Elworthy S, Ingham P, Whyte M . A transgenic zebrafish model of neutrophilic inflammation. Blood. 2006; 108(13):3976-8. DOI: 10.1182/blood-2006-05-024075. View

Pisharath H, Rhee J, Swanson M, Leach S, Parsons M . Targeted ablation of beta cells in the embryonic zebrafish pancreas using E. coli nitroreductase. Mech Dev. 2007; 124(3):218-29. PMC: 2583263. DOI: 10.1016/j.mod.2006.11.005. View

Kwan K, Fujimoto E, Grabher C, Mangum B, Hardy M, Campbell D . The Tol2kit: a multisite gateway-based construction kit for Tol2 transposon transgenesis constructs. Dev Dyn. 2007; 236(11):3088-99. DOI: 10.1002/dvdy.21343. View

Ellett F, Pase L, Hayman J, Andrianopoulos A, Lieschke G . mpeg1 promoter transgenes direct macrophage-lineage expression in zebrafish. Blood. 2010; 117(4):e49-56. PMC: 3056479. DOI: 10.1182/blood-2010-10-314120. View

Huang J, McKee M, Huang H, Xiang A, Davidson A, Lu H . A zebrafish model of conditional targeted podocyte ablation and regeneration. Kidney Int. 2013; 83(6):1193-200. PMC: 3672345. DOI: 10.1038/ki.2013.6. View

Hall C, Flores M, Storm T, Crosier K, Crosier P . The zebrafish lysozyme C promoter drives myeloid-specific expression in transgenic fish. BMC Dev Biol. 2007; 7:42. PMC: 1877083. DOI: 10.1186/1471-213X-7-42. View

Hall C, Sanderson L, Lawrence L, Pool B, van der Kroef M, Ashimbayeva E . Blocking fatty acid-fueled mROS production within macrophages alleviates acute gouty inflammation. J Clin Invest. 2018; 128(5):1752-1771. PMC: 5919807. DOI: 10.1172/JCI94584. View

Hsu C, Hou M, Hong J, Wu J, Her G . Inducible male infertility by targeted cell ablation in zebrafish testis. Mar Biotechnol (NY). 2009; 12(4):466-78. DOI: 10.1007/s10126-009-9248-4. View

10.

Lindmark D, Muller M . Antitrichomonad action, mutagenicity, and reduction of metronidazole and other nitroimidazoles. Antimicrob Agents Chemother. 1976; 10(3):476-82. PMC: 429775. DOI: 10.1128/AAC.10.3.476. View

11.

White D, Mumm J . The nitroreductase system of inducible targeted ablation facilitates cell-specific regenerative studies in zebrafish. Methods. 2013; 62(3):232-40. PMC: 3723733. DOI: 10.1016/j.ymeth.2013.03.017. View

12.

Edwards D . Nitroimidazole drugs--action and resistance mechanisms. II. Mechanisms of resistance. J Antimicrob Chemother. 1993; 31(2):201-10. DOI: 10.1093/jac/31.2.201. View

13.

Curado S, Stainier D, Anderson R . Nitroreductase-mediated cell/tissue ablation in zebrafish: a spatially and temporally controlled ablation method with applications in developmental and regeneration studies. Nat Protoc. 2008; 3(6):948-54. PMC: 2705989. DOI: 10.1038/nprot.2008.58. View

14.

Ellett F, Pazhakh V, Pase L, Benard E, Weerasinghe H, Azabdaftari D . Macrophages protect Talaromyces marneffei conidia from myeloperoxidase-dependent neutrophil fungicidal activity during infection establishment in vivo. PLoS Pathog. 2018; 14(6):e1007063. PMC: 6010348. DOI: 10.1371/journal.ppat.1007063. View

15.

Gregoire D, Kmita M . Genetic cell ablation. Methods Mol Biol. 2013; 1092:421-36. DOI: 10.1007/978-1-60327-292-6_25. View

16.

White Y, Woods D, Wood A . A transgenic zebrafish model of targeted oocyte ablation and de novo oogenesis. Dev Dyn. 2011; 240(8):1929-37. DOI: 10.1002/dvdy.22695. View

17.

Okuda K, Misa J, Oehlers S, Hall C, Ellett F, Alasmari S . A zebrafish model of inflammatory lymphangiogenesis. Biol Open. 2015; 4(10):1270-80. PMC: 4610225. DOI: 10.1242/bio.013540. View

18.

Curado S, Anderson R, Jungblut B, Mumm J, Schroeter E, Stainier D . Conditional targeted cell ablation in zebrafish: a new tool for regeneration studies. Dev Dyn. 2007; 236(4):1025-35. DOI: 10.1002/dvdy.21100. View

19.

Anlezark G, Melton R, Sherwood R, Coles B, Friedlos F, Knox R . The bioactivation of 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB1954)--I. Purification and properties of a nitroreductase enzyme from Escherichia coli--a potential enzyme for antibody-directed enzyme prodrug therapy (ADEPT). Biochem Pharmacol. 1992; 44(12):2289-95. DOI: 10.1016/0006-2952(92)90671-5. View