Metabolic profiling of tamoxifen-sensitive and tamoxifen-resistant breast cancer cells measuring the optical redox ratio of NAD(P)H/FAD utilizing a high-throughput time-resolved flow cytometer

Samantha Valentino; Kevin D. Houston; Jessica P. Houston

doi:10.1117/12.3002055

12 March 2024 Metabolic profiling of tamoxifen-sensitive and tamoxifen-resistant breast cancer cells measuring the optical redox ratio of NAD(P)H/FAD utilizing a high-throughput time-resolved flow cytometer

Samantha Valentino, Kevin D. Houston, Jessica P. Houston

Author Affiliations +

Proceedings Volume 12846, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XXII; 1284609 (2024) https://doi.org/10.1117/12.3002055
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Autofluorescent metabolic measurements of an optical redox ratio, NAD(P)H/FAD, have been utilized as a means of measuring cancer progression, treatment impact, subtype determination, and more. This optical redox ratio is traditionally measured through intensity with microscopy, but there is potential to adapt this technique for high throughput analysis using time-resolved flow cytometry with autofluorescence lifetime measurements. A fluorescence lifetime approach to these measurements allows for fluorophore concentration independent measurements that can provide new information to the field. The two fluorescent metabolites of interest that allow for redox analysis are NAD(P)H/NAD(P)⁺ and FADH₂/FAD. Variations in the redox state and binding of these metabolites to their respective coenzymes, have been correlated to the cycles in which cells metabolize glucose into ATP, either oxidative phosphorylation (OXPHOS) or glycolysis. This technique will be used herein to study the metabolism of MCF-7 tamoxifen resistant and sensitive breast cancer cells using flow cytometry, first on a fluorescence intensity basis of the two metabolites. Our results show there is a clear shift towards an increased redox ratio for tamoxifen resistant cells, indicating a greater reliance on glycolysis as a means of metabolism. Future work will focus on adapting the intensity based redox ratio approach through high-throughput flow cytometry used here to a fluorescence lifetime based measurement.

Conference Presentation

Citation Download Citation

Samantha Valentino, Kevin D. Houston, and Jessica P. Houston "Metabolic profiling of tamoxifen-sensitive and tamoxifen-resistant breast cancer cells measuring the optical redox ratio of NAD(P)H/FAD utilizing a high-throughput time-resolved flow cytometer", Proc. SPIE 12846, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XXII, 1284609 (12 March 2024); https://doi.org/10.1117/12.3002055

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