Factors Influencing Luciferase-Based Bioluminescent Imaging in Preclinical Models of Brain Tumor

Minjee Kim, Shiv K. Gupta, Wenjuan Zhang, Surabhi M Talele, Afroz Shareef Mohammad, Janice Laramy, Ann C. Mladek, Shuangling Zhang, Jann N Sarkaria, William F. Elmquist

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


Bioluminescent imaging (BLI) is a powerful tool in biomedical research to measure gene expression and tumor growth. The current study examined factors that influence the BLI signal, specifically focusing on the tissue distribution of two luciferase substrates, D-luciferin and CycLuc1. D-luciferin, a natural substrate of firefly luciferase, has been reported to have limited brain distribution, possibly due to the efflux transporter, breast cancer resistance protein (Bcrp), at the blood-brain barrier. CycLuc1, a synthetic analog of D-luciferin, has a greater BLI signal at lower doses than D-luciferin, especially in the brain. Our results indicate that limited brain distribution of D-luciferin and CycLuc1 is predominantly dictated by their low intrinsic permeability across the cell membrane, where the efflux transporter, Bcrp, plays a relatively minor role. Both genetic ablation and pharmacological inhibition of Bcrp decreased the systemic clearance of both luciferase substrates, significantly increasing exposure in the blood and, hence, in organs and tissues. These data also indicate that the biodistribution of luciferase substrates can be differentially influenced in luciferase-bearing tissues, leading to a “tissue-dependent” BLI signal. The results of this study point to the need to consider multiple mechanisms that influence the distribution of luciferase substrates. SIGNIFICANCE STATEMENT Bioluminescence is used to monitor many biological processes, including tumor growth. This study examined the pharmacokinetics, brain distribution, and the role of active efflux transporters on the luciferase substrates D-luciferin and CycLuc1. CycLuc1 has a more sustained systemic circulation time (longer half-life) that can provide an advantage for the superior imaging outcome of CycLuc1 over D-luciferin. The disparity in imaging intensities between brain and peripheral sites is due to low intrinsic permeability of these luciferase substrates across the blood-brain barrier.

Original languageEnglish (US)
Pages (from-to)277-286
Number of pages10
JournalDrug Metabolism and Disposition
Issue number3
StatePublished - Mar 1 2022

Bibliographical note

Funding Information:
This work was supported by National Institutes of Health National Cancer Institute [Grants RO1-CA138437, U54-CA210181, U01-CA227954, and P50-CA108960] and National Institutes of Health National Institute of Neurological Disorders and Stroke [Grant RO1 NS077921]. No author has an actual or perceived conflict of interest with the contents of this article. dx.doi.org/10.1124/dmd.121.000597. S This article has supplemental material available at dmd.aspetjournals.org.

Publisher Copyright:
Copyright © 2022 by The American Society for Pharmacology and Experimental Therapeutics


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