title: Continuous, real-time bioimaging of chemical bioavailability and toxicology using autonomously bioluminescent human cell lines.
abstract: Bioluminescent imaging is an emerging biomedical surveillance strategy that uses external cameras to detectin vivolight generated in small animal models of human physiology orin vitrolight generated in tissue culture or tissue scaffold mimics of human anatomy. The most widely utilized of reporters is the firefly luciferase (luc) gene; however, it generates light only upon addition of a chemical substrate, thus only generating intermittent single time point data snapshots. To overcome this disadvantage, we have demonstrated substrate-independent bioluminescent imaging using an optimized bacterial bioluminescence (lux) system. Theluxreporter produces bioluminescence autonomously using components found naturally within the cell, thereby allowing imaging to occur continuously and in real-time over the lifetime of the host. We have validated this technology in human cells with demonstrated chemical toxicological profiling against exotoxin exposures at signal strengths comparable to existinglucsystems (~1.33 × 107photons/second). As a proof-in-principle demonstration, we have engineered breast carcinoma cells to express bioluminescence for real-time screening of endocrine disrupting chemicals and validated detection of 17β-estradiol (EC50= ~ 10 pM). These and other applications of this new reporter technology will be discussed as potential new pathways towards improved models of target chemical bioavailability, toxicology, efficacy, and human safety.
authors.affiliation: Joint Institute for Biological Sciences, The University of Tennessee, 676 Dabney Hall, Knoxville, TN, USA 37996.
authors.forename: Tingting
authors.initials: T
authors.lastname: Xu
authors.affiliation: 490 BioTech Inc., 2450 E. J. Chapman Drive, Knoxville, TN USA 37996.
authors.forename: Dan M
authors.initials: DM
authors.lastname: Close
authors.affiliation: Center for Environmental Biotechnology, The University of Tennessee, 676 Dabney Hall, Knoxville, TN, USA 37996.
authors.forename: James D
authors.initials: JD
authors.lastname: Webb
authors.affiliation: Center for Environmental Biotechnology, The University of Tennessee, 676 Dabney Hall, Knoxville, TN, USA 37996.
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Price
authors.affiliation: 490 BioTech Inc., 2450 E. J. Chapman Drive, Knoxville, TN USA 37996 ; Center for Environmental Biotechnology, The University of Tennessee, 676 Dabney Hall, Knoxville, TN, USA 37996.
authors.forename: Steven A
authors.initials: SA
authors.lastname: Ripp
authors.affiliation: Joint Institute for Biological Sciences, The University of Tennessee, 676 Dabney Hall, Knoxville, TN, USA 37996 ; 490 BioTech Inc., 2450 E. J. Chapman Drive, Knoxville, TN USA 37996 ; Center for Environmental Biotechnology, The University of Tennessee, 676 Dabney Hall, Knoxville, TN, USA 37996.
authors.forename: Gary S
authors.initials: GS
authors.lastname: Sayler
keywords: Bioavailability; Bioluminescent imaging; Bioreporter; Estrogen; Luciferase; Lux; Tissue scaffold; Toxicology