Internally controlled and dynamic optical measures of functional tumor biology

Chung, T., Garcia, L., Swamynathan, M. M., Froeling, F. E.M. , Trotman, L. C., Tuveson, D. A. and Lyons, S. K. (2023) Internally controlled and dynamic optical measures of functional tumor biology. Analytical Chemistry, 95(13), pp. 5661-5670. (doi: 10.1021/acs.analchem.2c05450) (PMID:36952386) (PMCID:PMC10077328)

Abstract

Imaging defined aspects of functional tumor biology with bioluminescent reporter transgenes is a popular approach in preclinical drug development as it is sensitive, relatively high-throughput and low cost. However, the lack of internal controls subject functional bioluminescence to a number of unpredictable variables that reduce this powerful tool to semi-quantitative interpretation of large-scale effects. Here, we report the generation of sensitive and quantitative live reporters for two key measures of functional cancer biology and pharmacologic stress: the cell cycle and oxidative stress. We developed a two-colored readout, where two independent enzymes convert a common imaging substrate into spectrally distinguishable light. The signal intensity of one color is dependent upon the biological state, whereas the other color is constitutively expressed. The ratio of emitted colored light corrects the functional signal for independent procedural variables, substantially improving the robustness and interpretation of relatively low-fold changes in functional signal intensity after drug treatment. The application of these readouts in vitro is highly advantageous, as peak cell response to therapy can now be readily visualized for single or combination treatments and not simply assessed at an arbitrary and destructive timepoint. Spectral imaging in vivo can be challenging, but we also present evidence to show that the reporters can work in this context as well. Collectively, the development and validation of these internally controlled reporters allow researchers to robustly and dynamically visualize tumor cell biology in response to treatment. Given the prevalence of bioluminescence imaging, this presents significant and much needed opportunities for preclinical therapeutic development.

• Item Type: Articles
• Additional Information: The authors acknowledge support by Developmental Funds from the Cancer Center Support grant 5P30CA045508 and from the Cold Spring Harbor Laboratory and Northwell Health Affiliation. This work was also performed with assistance from the CSHL Flow Cytometry and Animal Shared Resources, which are supported in part by the Cancer Center Support grant 5P30CA045508.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Froeling, Dr Fieke
• Authors: Chung, T., Garcia, L., Swamynathan, M. M., Froeling, F. E.M., Trotman, L. C., Tuveson, D. A., and Lyons, S. K.
• College/School: College of Medical Veterinary and Life Sciences > School of Cancer Sciences
• Journal Name: Analytical Chemistry
• Publisher: American Chemical Society
• ISSN: 0003-2700
• ISSN (Online): 1520-6882
• Published Online: 23 March 2023
• Copyright Holders: Copyright © 2023 The Authors
• First Published: First published in Analytical Chemistry 95(13): 5661-5670
• Publisher Policy: Reproduced under a Creative Commons License