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Simultaneous Probing of Metabolism and Oxygenation of Tumors In Vivo Using FLIM of NAD(P)H and PLIM of a New Polymeric Ir(III) Oxygen Sensor
Tumor cells are well adapted to grow in conditions of variable oxygen supply and hypoxia by switching between different metabolic pathways. However, the regulatory effect of oxygen on metabolism and its contribution to the metabolic heterogeneity of tumors have not been fully explored. In this study...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9499414/ https://www.ncbi.nlm.nih.gov/pubmed/36142177 http://dx.doi.org/10.3390/ijms231810263 |
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author | Parshina, Yulia P. Komarova, Anastasia D. Bochkarev, Leonid N. Kovylina, Tatyana A. Plekhanov, Anton A. Klapshina, Larisa G. Konev, Aleksey N. Mozherov, Artem M. Shchechkin, Ilya D. Sirotkina, Marina A. Shcheslavskiy, Vladislav I. Shirmanova, Marina V. |
author_facet | Parshina, Yulia P. Komarova, Anastasia D. Bochkarev, Leonid N. Kovylina, Tatyana A. Plekhanov, Anton A. Klapshina, Larisa G. Konev, Aleksey N. Mozherov, Artem M. Shchechkin, Ilya D. Sirotkina, Marina A. Shcheslavskiy, Vladislav I. Shirmanova, Marina V. |
author_sort | Parshina, Yulia P. |
collection | PubMed |
description | Tumor cells are well adapted to grow in conditions of variable oxygen supply and hypoxia by switching between different metabolic pathways. However, the regulatory effect of oxygen on metabolism and its contribution to the metabolic heterogeneity of tumors have not been fully explored. In this study, we develop a methodology for the simultaneous analysis of cellular metabolic status, using the fluorescence lifetime imaging microscopy (FLIM) of metabolic cofactor NAD(P)H, and oxygen level, using the phosphorescence lifetime imaging (PLIM) of a new polymeric Ir(III)-based sensor (PIr3) in tumors in vivo. The sensor, derived from a polynorbornene and cyclometalated iridium(III) complex, exhibits the oxygen-dependent quenching of phosphorescence with a 40% longer lifetime in degassed compared to aerated solutions. In vitro, hypoxia resulted in a correlative increase in PIr3 phosphorescence lifetime and free (glycolytic) NAD(P)H fraction in cells. In vivo, mouse tumors demonstrated a high degree of cellular-level heterogeneity of both metabolic and oxygen states, and a lower dependence of metabolism on oxygen than cells in vitro. The small tumors were hypoxic, while the advanced tumors contained areas of normoxia and hypoxia, which was consistent with the pimonidazole assay and angiographic imaging. Dual FLIM/PLIM metabolic/oxygen imaging will be valuable in preclinical investigations into the effects of hypoxia on metabolic aspects of tumor progression and treatment response. |
format | Online Article Text |
id | pubmed-9499414 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-94994142022-09-23 Simultaneous Probing of Metabolism and Oxygenation of Tumors In Vivo Using FLIM of NAD(P)H and PLIM of a New Polymeric Ir(III) Oxygen Sensor Parshina, Yulia P. Komarova, Anastasia D. Bochkarev, Leonid N. Kovylina, Tatyana A. Plekhanov, Anton A. Klapshina, Larisa G. Konev, Aleksey N. Mozherov, Artem M. Shchechkin, Ilya D. Sirotkina, Marina A. Shcheslavskiy, Vladislav I. Shirmanova, Marina V. Int J Mol Sci Article Tumor cells are well adapted to grow in conditions of variable oxygen supply and hypoxia by switching between different metabolic pathways. However, the regulatory effect of oxygen on metabolism and its contribution to the metabolic heterogeneity of tumors have not been fully explored. In this study, we develop a methodology for the simultaneous analysis of cellular metabolic status, using the fluorescence lifetime imaging microscopy (FLIM) of metabolic cofactor NAD(P)H, and oxygen level, using the phosphorescence lifetime imaging (PLIM) of a new polymeric Ir(III)-based sensor (PIr3) in tumors in vivo. The sensor, derived from a polynorbornene and cyclometalated iridium(III) complex, exhibits the oxygen-dependent quenching of phosphorescence with a 40% longer lifetime in degassed compared to aerated solutions. In vitro, hypoxia resulted in a correlative increase in PIr3 phosphorescence lifetime and free (glycolytic) NAD(P)H fraction in cells. In vivo, mouse tumors demonstrated a high degree of cellular-level heterogeneity of both metabolic and oxygen states, and a lower dependence of metabolism on oxygen than cells in vitro. The small tumors were hypoxic, while the advanced tumors contained areas of normoxia and hypoxia, which was consistent with the pimonidazole assay and angiographic imaging. Dual FLIM/PLIM metabolic/oxygen imaging will be valuable in preclinical investigations into the effects of hypoxia on metabolic aspects of tumor progression and treatment response. MDPI 2022-09-06 /pmc/articles/PMC9499414/ /pubmed/36142177 http://dx.doi.org/10.3390/ijms231810263 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Parshina, Yulia P. Komarova, Anastasia D. Bochkarev, Leonid N. Kovylina, Tatyana A. Plekhanov, Anton A. Klapshina, Larisa G. Konev, Aleksey N. Mozherov, Artem M. Shchechkin, Ilya D. Sirotkina, Marina A. Shcheslavskiy, Vladislav I. Shirmanova, Marina V. Simultaneous Probing of Metabolism and Oxygenation of Tumors In Vivo Using FLIM of NAD(P)H and PLIM of a New Polymeric Ir(III) Oxygen Sensor |
title | Simultaneous Probing of Metabolism and Oxygenation of Tumors In Vivo Using FLIM of NAD(P)H and PLIM of a New Polymeric Ir(III) Oxygen Sensor |
title_full | Simultaneous Probing of Metabolism and Oxygenation of Tumors In Vivo Using FLIM of NAD(P)H and PLIM of a New Polymeric Ir(III) Oxygen Sensor |
title_fullStr | Simultaneous Probing of Metabolism and Oxygenation of Tumors In Vivo Using FLIM of NAD(P)H and PLIM of a New Polymeric Ir(III) Oxygen Sensor |
title_full_unstemmed | Simultaneous Probing of Metabolism and Oxygenation of Tumors In Vivo Using FLIM of NAD(P)H and PLIM of a New Polymeric Ir(III) Oxygen Sensor |
title_short | Simultaneous Probing of Metabolism and Oxygenation of Tumors In Vivo Using FLIM of NAD(P)H and PLIM of a New Polymeric Ir(III) Oxygen Sensor |
title_sort | simultaneous probing of metabolism and oxygenation of tumors in vivo using flim of nad(p)h and plim of a new polymeric ir(iii) oxygen sensor |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9499414/ https://www.ncbi.nlm.nih.gov/pubmed/36142177 http://dx.doi.org/10.3390/ijms231810263 |
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