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Tracking Functional Tumor Cell Subpopulations of Malignant Glioma by Phasor Fluorescence Lifetime Imaging Microscopy of NADH

Intra-tumoral heterogeneity is associated with therapeutic resistance of cancer and there exists a need to non-invasively identify functional tumor subpopulations responsible for tumor recurrence. Reduced nicotinamide adenine dinucleotide (NADH) is a metabolic coenzyme essential in cellular respirat...

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Autores principales: Trinh, Andrew L., Chen, Hongtao, Chen, Yumay, Hu, Yuanjie, Li, Zhenzhi, Siegel, Eric R., Linskey, Mark E., Wang, Ping H., Digman, Michelle A., Zhou, Yi-Hong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5742816/
https://www.ncbi.nlm.nih.gov/pubmed/29211022
http://dx.doi.org/10.3390/cancers9120168
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author Trinh, Andrew L.
Chen, Hongtao
Chen, Yumay
Hu, Yuanjie
Li, Zhenzhi
Siegel, Eric R.
Linskey, Mark E.
Wang, Ping H.
Digman, Michelle A.
Zhou, Yi-Hong
author_facet Trinh, Andrew L.
Chen, Hongtao
Chen, Yumay
Hu, Yuanjie
Li, Zhenzhi
Siegel, Eric R.
Linskey, Mark E.
Wang, Ping H.
Digman, Michelle A.
Zhou, Yi-Hong
author_sort Trinh, Andrew L.
collection PubMed
description Intra-tumoral heterogeneity is associated with therapeutic resistance of cancer and there exists a need to non-invasively identify functional tumor subpopulations responsible for tumor recurrence. Reduced nicotinamide adenine dinucleotide (NADH) is a metabolic coenzyme essential in cellular respiration. Fluorescence lifetime imaging microscopy (FLIM) of NADH has been demonstrated to be a powerful label-free indicator for inferring metabolic states of living cells. Using FLIM, we identified a significant shift towards longer NADH fluorescence lifetimes, suggesting an increase in the fraction of protein-bound NADH, in the invasive stem-like tumor-initiating cell (STIC) subpopulation relative to the tumor mass-forming cell (TMC) subpopulation of malignant gliomas. By applying our previously studied model to transition glioma from a majority of STIC to a majority of TMC in serum-adherent culture conditions following serial passages, we compared changes in NADH states, cellular respirations (oxidative phosphorylation and glycolysis), EGFR expression, and cell-growth speed over passages. We identified a significant positive correlation between free-NADH fraction and cell growth, which was related to an increase of TMC fraction. In comparison, the increase of EGFR and cellular respirations preceded all these changes. In conclusion, FLIM of NADH provides a non-invasive method to monitor the dynamics of tumor heterogeneity before and after treatment.
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spelling pubmed-57428162017-12-29 Tracking Functional Tumor Cell Subpopulations of Malignant Glioma by Phasor Fluorescence Lifetime Imaging Microscopy of NADH Trinh, Andrew L. Chen, Hongtao Chen, Yumay Hu, Yuanjie Li, Zhenzhi Siegel, Eric R. Linskey, Mark E. Wang, Ping H. Digman, Michelle A. Zhou, Yi-Hong Cancers (Basel) Article Intra-tumoral heterogeneity is associated with therapeutic resistance of cancer and there exists a need to non-invasively identify functional tumor subpopulations responsible for tumor recurrence. Reduced nicotinamide adenine dinucleotide (NADH) is a metabolic coenzyme essential in cellular respiration. Fluorescence lifetime imaging microscopy (FLIM) of NADH has been demonstrated to be a powerful label-free indicator for inferring metabolic states of living cells. Using FLIM, we identified a significant shift towards longer NADH fluorescence lifetimes, suggesting an increase in the fraction of protein-bound NADH, in the invasive stem-like tumor-initiating cell (STIC) subpopulation relative to the tumor mass-forming cell (TMC) subpopulation of malignant gliomas. By applying our previously studied model to transition glioma from a majority of STIC to a majority of TMC in serum-adherent culture conditions following serial passages, we compared changes in NADH states, cellular respirations (oxidative phosphorylation and glycolysis), EGFR expression, and cell-growth speed over passages. We identified a significant positive correlation between free-NADH fraction and cell growth, which was related to an increase of TMC fraction. In comparison, the increase of EGFR and cellular respirations preceded all these changes. In conclusion, FLIM of NADH provides a non-invasive method to monitor the dynamics of tumor heterogeneity before and after treatment. MDPI 2017-12-06 /pmc/articles/PMC5742816/ /pubmed/29211022 http://dx.doi.org/10.3390/cancers9120168 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Trinh, Andrew L.
Chen, Hongtao
Chen, Yumay
Hu, Yuanjie
Li, Zhenzhi
Siegel, Eric R.
Linskey, Mark E.
Wang, Ping H.
Digman, Michelle A.
Zhou, Yi-Hong
Tracking Functional Tumor Cell Subpopulations of Malignant Glioma by Phasor Fluorescence Lifetime Imaging Microscopy of NADH
title Tracking Functional Tumor Cell Subpopulations of Malignant Glioma by Phasor Fluorescence Lifetime Imaging Microscopy of NADH
title_full Tracking Functional Tumor Cell Subpopulations of Malignant Glioma by Phasor Fluorescence Lifetime Imaging Microscopy of NADH
title_fullStr Tracking Functional Tumor Cell Subpopulations of Malignant Glioma by Phasor Fluorescence Lifetime Imaging Microscopy of NADH
title_full_unstemmed Tracking Functional Tumor Cell Subpopulations of Malignant Glioma by Phasor Fluorescence Lifetime Imaging Microscopy of NADH
title_short Tracking Functional Tumor Cell Subpopulations of Malignant Glioma by Phasor Fluorescence Lifetime Imaging Microscopy of NADH
title_sort tracking functional tumor cell subpopulations of malignant glioma by phasor fluorescence lifetime imaging microscopy of nadh
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5742816/
https://www.ncbi.nlm.nih.gov/pubmed/29211022
http://dx.doi.org/10.3390/cancers9120168
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