Pseudomonas aeruginosa NfsB and nitro-CBI-DEI – a promising enzyme-prodrug combination for gene directed enzyme prodrug therapyReportar como inadecuado




Pseudomonas aeruginosa NfsB and nitro-CBI-DEI – a promising enzyme-prodrug combination for gene directed enzyme prodrug therapy - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Molecular Cancer

, 12:58

First Online: 10 June 2013Received: 08 April 2013Accepted: 05 June 2013DOI: 10.1186-1476-4598-12-58

Cite this article as: Green, L.K., Syddall, S.P., Carlin, K.M. et al. Mol Cancer 2013 12: 58. doi:10.1186-1476-4598-12-58

Abstract

BackgroundThe nitro-chloromethylbenzindoline prodrug nitro-CBI-DEI appears a promising candidate for the anti-cancer strategy gene-directed enzyme prodrug therapy, based on its ability to be converted to a highly cytotoxic cell-permeable derivative by the nitroreductase NfsB from Escherichia coli. However, relative to some other nitroaromatic prodrugs, nitro-CBI-DEI is a poor substrate for E. coli NfsB. To address this limitation we evaluated other nitroreductase candidates from E. coli and Pseudomonas aeruginosa.

FindingsInitial screens of candidate genes in the E. coli reporter strain SOS-R2 identified two additional nitroreductases, E. coli NfsA and P. aeruginosa NfsB, as being more effective activators of nitro-CBI-DEI than E. coli NfsB. In monolayer cytotoxicity assays, human colon carcinoma HCT-116 cells transfected with P. aeruginosa NfsB were >4.5-fold more sensitive to nitro-CBI-DEI than cells expressing either E. coli enzyme, and 23.5-fold more sensitive than untransfected HCT-116. In three dimensional mixed cell cultures, not only were the P. aeruginosa NfsB expressing cells 540-fold more sensitive to nitro-CBI-DEI than pure cultures of untransfected HCT-116, the activated drug that they generated also displayed an unprecedented local bystander effect.

ConclusionWe posit that the discrepancy in the fold-sensitivity to nitro-CBI-DEI between the two and three dimensional cytotoxicity assays stems from loss of activated drug into the media in the monolayer cultures. This emphasises the importance of evaluating high-bystander GDEPT prodrugs in three dimensional models. The high cytotoxicity and bystander effect exhibited by the NfsB Pa-nitro-CBI-DEI combination suggest that further preclinical development of this GDEPT pairing is warranted.

KeywordsGene therapy GDEPT Nitroaromatic prodrug Nitroreductase Nitro-CBI-DEI CB1954 SOS chromotest Bystander effect Electronic supplementary materialThe online version of this article doi:10.1186-1476-4598-12-58 contains supplementary material, which is available to authorized users.

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Autor: Laura K Green - Sophie P Syddall - Kendall M Carlin - Glenn D Bell - Christopher P Guise - Alexandra M Mowday - Michae

Fuente: https://link.springer.com/







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