Internally Controlled and Dynamic Optical Measures of Functional Tumor Biology

Chung, Taemoon, Garcia, Libia, Swamynathan, Manojit M, Froeling, Fieke EM, Trotman, Lloyd C, Tuveson, David A, Lyons, Scott K (April 2023) Internally Controlled and Dynamic Optical Measures of Functional Tumor Biology. Analytical Chemistry, 95 (13). pp. 5661-5670. ISSN 0003-2700

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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: Paper
Subjects: diseases & disorders > cancer
diseases & disorders
diseases & disorders > pulmonary disease > oxidative stress
CSHL Authors:
Communities: CSHL labs > Lyons lab
CSHL labs > Trotman lab
CSHL labs > Tuveson lab
CSHL Cancer Center Program
CSHL Cancer Center Program > Cellular Communication in Cancer Program
CSHL Cancer Center Shared Resources > Animal Services
CSHL Cancer Center Shared Resources > Bioinformatics Service
CSHL Cancer Center Shared Resources > Flow Cytometry Service
CSHL Cancer Center Shared Resources > Animal Shared Resource
SWORD Depositor: CSHL Elements
Depositing User: CSHL Elements
Date: 4 April 2023
Date Deposited: 25 Sep 2023 15:26
Last Modified: 09 Feb 2024 14:11
PMCID: PMC10077328
Related URLs:
URI: https://repository.cshl.edu/id/eprint/40889

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