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Title: Co-Cultured Continuously Bioluminescent Human Cells as Bioreporters for the Detection of Prodrug Therapeutic Impact Pre- and Post-Metabolism

Abstract

Modern drug discovery workflows require assay systems capable of replicating the complex interactions of multiple tissue types, but that can still function under high throughput conditions. In this work, we evaluate the use of substrate-free autobioluminescence in human cell lines to support the performance of these assays with reduced economical and logistical restrictions relative to substrate-requiring bioluminescent reporter systems. The use of autobioluminescence was found to support assay functionality similar to existing luciferase reporter targets. The autobioluminescent assay format was observed to correlate strongly with general metabolic activity markers such as ATP content and the presence of reactive oxygen species, but not with secondary markers such as glutathione depletion. At the transcriptional level, autobioluminescent dynamics were most closely associated with expression of the CYP1A1 phase I detoxification pathway. These results suggest constitutively autobioluminescent cells can function as general metabolic activity bioreporters, while pairing expression of the autobioluminescent phenotype to detoxification pathway specific promoters could create more specific sensor systems.

Authors:
 [1];  [2];  [3];  [4];  [2];  [2]
  1. Univ. of Tennessee, Knoxville, TN (United States)
  2. 490 BioTech, Inc., Knoxville, TN (United States)
  3. Cornell Univ., Ithaca, NY (United States)
  4. Univ. of Tennessee, Knoxville, TN (United States); 490 BioTech, Inc., Knoxville, TN (United States)
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1476398
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Accepted Manuscript
Journal Name:
Sensors
Additional Journal Information:
Journal Volume: 17; Journal Issue: 12; Journal ID: ISSN 1424-8220
Publisher:
MDPI AG
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Xu, Tingting, Conway, Michael, Frank, Ashley, Ripp, Steven, Sayler, Gary, and Close, Dan. Co-Cultured Continuously Bioluminescent Human Cells as Bioreporters for the Detection of Prodrug Therapeutic Impact Pre- and Post-Metabolism. United States: N. p., 2017. Web. doi:10.3390/s17122827.
Xu, Tingting, Conway, Michael, Frank, Ashley, Ripp, Steven, Sayler, Gary, & Close, Dan. Co-Cultured Continuously Bioluminescent Human Cells as Bioreporters for the Detection of Prodrug Therapeutic Impact Pre- and Post-Metabolism. United States. doi:10.3390/s17122827.
Xu, Tingting, Conway, Michael, Frank, Ashley, Ripp, Steven, Sayler, Gary, and Close, Dan. Wed . "Co-Cultured Continuously Bioluminescent Human Cells as Bioreporters for the Detection of Prodrug Therapeutic Impact Pre- and Post-Metabolism". United States. doi:10.3390/s17122827. https://www.osti.gov/servlets/purl/1476398.
@article{osti_1476398,
title = {Co-Cultured Continuously Bioluminescent Human Cells as Bioreporters for the Detection of Prodrug Therapeutic Impact Pre- and Post-Metabolism},
author = {Xu, Tingting and Conway, Michael and Frank, Ashley and Ripp, Steven and Sayler, Gary and Close, Dan},
abstractNote = {Modern drug discovery workflows require assay systems capable of replicating the complex interactions of multiple tissue types, but that can still function under high throughput conditions. In this work, we evaluate the use of substrate-free autobioluminescence in human cell lines to support the performance of these assays with reduced economical and logistical restrictions relative to substrate-requiring bioluminescent reporter systems. The use of autobioluminescence was found to support assay functionality similar to existing luciferase reporter targets. The autobioluminescent assay format was observed to correlate strongly with general metabolic activity markers such as ATP content and the presence of reactive oxygen species, but not with secondary markers such as glutathione depletion. At the transcriptional level, autobioluminescent dynamics were most closely associated with expression of the CYP1A1 phase I detoxification pathway. These results suggest constitutively autobioluminescent cells can function as general metabolic activity bioreporters, while pairing expression of the autobioluminescent phenotype to detoxification pathway specific promoters could create more specific sensor systems.},
doi = {10.3390/s17122827},
journal = {Sensors},
number = 12,
volume = 17,
place = {United States},
year = {2017},
month = {12}
}

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