Correlating NAD(P)H lifetime shifts to tamoxifen resistance in breast cancer cells: A metabolic screening study with time-resolved flow cytometry
Time-resolved flow cytometry (TRFC) was used to measure metabolic differences in estrogen receptor-positive breast cancer cells. This specialty cytometry technique measures fluorescence lifetimes as a single-cell parameter thereby providing a unique approach for high-throughput cell counting and scr...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
World Scientific Publishing
2025-01-01
|
| Series: | Journal of Innovative Optical Health Sciences |
| Subjects: | |
| Online Access: | https://www.worldscientific.com/doi/10.1142/S1793545824500202 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832585067086479360 |
|---|---|
| author | Samantha Valentino Karla Ortega-Sandoval Kevin D. Houston Jessica P. Houston |
| author_facet | Samantha Valentino Karla Ortega-Sandoval Kevin D. Houston Jessica P. Houston |
| author_sort | Samantha Valentino |
| collection | DOAJ |
| description | Time-resolved flow cytometry (TRFC) was used to measure metabolic differences in estrogen receptor-positive breast cancer cells. This specialty cytometry technique measures fluorescence lifetimes as a single-cell parameter thereby providing a unique approach for high-throughput cell counting and screening. Differences in fluorescence lifetime were detected and this was associated with sensitivity to the commonly prescribed therapeutic tamoxifen. Differences in fluorescence lifetime are attributed to the binding states of the autofluorescent metabolite NAD(P)H. The function of NAD(P)H is well described and in general involves cycling from a reduced to oxidized state to facilitate electron transport for the conversion of pyruvate to lactate. NAD(P)H fluorescence lifetimes depend on the bound or unbound state of the metabolite, which also relates to metabolic transitions between oxidative phosphorylation and glycolysis. To determine if fundamental metabolic profiles differ for cells that are sensitive to tamoxifen compared to those that are resistant, large populations of MCF-7 breast cancer cells were screened and fluorescence lifetimes were quantified. Additionally, metabolic differences associated with tamoxifen sensitivity were measured with a Seahorse HS mini metabolic analyzer (Agilent Technologies Inc. Santa Clara, CA) and confocal imaging. Results show that tamoxifen-resistant breast cancer cells have increased utilization of glycolysis for energy production compared to tamoxifen-sensitive breast cancer cells. This work is impacting because it establishes an early step toward developing a reliable screening technology in which large cell censuses can be differentiated for drug sensitivity in a label-free fashion. |
| format | Article |
| id | doaj-art-5c6f938ebf7f40858bfc3330ea2776b1 |
| institution | Kabale University |
| issn | 1793-5458 1793-7205 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | World Scientific Publishing |
| record_format | Article |
| series | Journal of Innovative Optical Health Sciences |
| spelling | doaj-art-5c6f938ebf7f40858bfc3330ea2776b12025-01-27T05:49:52ZengWorld Scientific PublishingJournal of Innovative Optical Health Sciences1793-54581793-72052025-01-01180110.1142/S1793545824500202Correlating NAD(P)H lifetime shifts to tamoxifen resistance in breast cancer cells: A metabolic screening study with time-resolved flow cytometrySamantha Valentino0Karla Ortega-Sandoval1Kevin D. Houston2Jessica P. Houston3Chemical and Materials Engineering, New Mexico State University, 1040 S Horseshoe Dr., Las Cruces, NM 88003, USAChemical and Materials Engineering, New Mexico State University, 1040 S Horseshoe Dr., Las Cruces, NM 88003, USAChemistry and Biochemistry, New Mexico State University, 1175 N Horseshoe Dr., Las Cruces, NM 88003, USAChemical and Materials Engineering, New Mexico State University, 1040 S Horseshoe Dr., Las Cruces, NM 88003, USATime-resolved flow cytometry (TRFC) was used to measure metabolic differences in estrogen receptor-positive breast cancer cells. This specialty cytometry technique measures fluorescence lifetimes as a single-cell parameter thereby providing a unique approach for high-throughput cell counting and screening. Differences in fluorescence lifetime were detected and this was associated with sensitivity to the commonly prescribed therapeutic tamoxifen. Differences in fluorescence lifetime are attributed to the binding states of the autofluorescent metabolite NAD(P)H. The function of NAD(P)H is well described and in general involves cycling from a reduced to oxidized state to facilitate electron transport for the conversion of pyruvate to lactate. NAD(P)H fluorescence lifetimes depend on the bound or unbound state of the metabolite, which also relates to metabolic transitions between oxidative phosphorylation and glycolysis. To determine if fundamental metabolic profiles differ for cells that are sensitive to tamoxifen compared to those that are resistant, large populations of MCF-7 breast cancer cells were screened and fluorescence lifetimes were quantified. Additionally, metabolic differences associated with tamoxifen sensitivity were measured with a Seahorse HS mini metabolic analyzer (Agilent Technologies Inc. Santa Clara, CA) and confocal imaging. Results show that tamoxifen-resistant breast cancer cells have increased utilization of glycolysis for energy production compared to tamoxifen-sensitive breast cancer cells. This work is impacting because it establishes an early step toward developing a reliable screening technology in which large cell censuses can be differentiated for drug sensitivity in a label-free fashion.https://www.worldscientific.com/doi/10.1142/S1793545824500202Time-resolvedflow cytometryautofluorescencefluorescence lifetimebreast cancermetabolism |
| spellingShingle | Samantha Valentino Karla Ortega-Sandoval Kevin D. Houston Jessica P. Houston Correlating NAD(P)H lifetime shifts to tamoxifen resistance in breast cancer cells: A metabolic screening study with time-resolved flow cytometry Journal of Innovative Optical Health Sciences Time-resolved flow cytometry autofluorescence fluorescence lifetime breast cancer metabolism |
| title | Correlating NAD(P)H lifetime shifts to tamoxifen resistance in breast cancer cells: A metabolic screening study with time-resolved flow cytometry |
| title_full | Correlating NAD(P)H lifetime shifts to tamoxifen resistance in breast cancer cells: A metabolic screening study with time-resolved flow cytometry |
| title_fullStr | Correlating NAD(P)H lifetime shifts to tamoxifen resistance in breast cancer cells: A metabolic screening study with time-resolved flow cytometry |
| title_full_unstemmed | Correlating NAD(P)H lifetime shifts to tamoxifen resistance in breast cancer cells: A metabolic screening study with time-resolved flow cytometry |
| title_short | Correlating NAD(P)H lifetime shifts to tamoxifen resistance in breast cancer cells: A metabolic screening study with time-resolved flow cytometry |
| title_sort | correlating nad p h lifetime shifts to tamoxifen resistance in breast cancer cells a metabolic screening study with time resolved flow cytometry |
| topic | Time-resolved flow cytometry autofluorescence fluorescence lifetime breast cancer metabolism |
| url | https://www.worldscientific.com/doi/10.1142/S1793545824500202 |
| work_keys_str_mv | AT samanthavalentino correlatingnadphlifetimeshiftstotamoxifenresistanceinbreastcancercellsametabolicscreeningstudywithtimeresolvedflowcytometry AT karlaortegasandoval correlatingnadphlifetimeshiftstotamoxifenresistanceinbreastcancercellsametabolicscreeningstudywithtimeresolvedflowcytometry AT kevindhouston correlatingnadphlifetimeshiftstotamoxifenresistanceinbreastcancercellsametabolicscreeningstudywithtimeresolvedflowcytometry AT jessicaphouston correlatingnadphlifetimeshiftstotamoxifenresistanceinbreastcancercellsametabolicscreeningstudywithtimeresolvedflowcytometry |