Measurement of Patient-Derived Glioblastoma Cell Response to Temozolomide Using Fluorescence Lifetime Imaging of NAD(P)H
Personalized strategies in glioblastoma treatment are highly necessary. One of the possible approaches is drug screening using patient-derived tumor cells. However, this requires reliable methods for assessment of the response of tumor cells to treatment. Fluorescence lifetime imaging microscopy (FL...
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| Format: | Article |
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MDPI AG
2023-05-01
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| Series: | Pharmaceuticals |
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| Online Access: | https://www.mdpi.com/1424-8247/16/6/796 |
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| author | Diana V. Yuzhakova Daria A. Sachkova Marina V. Shirmanova Artem M. Mozherov Anna V. Izosimova Anna S. Zolotova Konstantin S. Yashin |
| author_facet | Diana V. Yuzhakova Daria A. Sachkova Marina V. Shirmanova Artem M. Mozherov Anna V. Izosimova Anna S. Zolotova Konstantin S. Yashin |
| author_sort | Diana V. Yuzhakova |
| collection | DOAJ |
| description | Personalized strategies in glioblastoma treatment are highly necessary. One of the possible approaches is drug screening using patient-derived tumor cells. However, this requires reliable methods for assessment of the response of tumor cells to treatment. Fluorescence lifetime imaging microscopy (FLIM) is a promising instrument to detect early cellular response to chemotherapy using the autofluorescence of metabolic cofactors. Here, we explored FLIM of NAD(P)H to evaluate the sensitivity of patient-derived glioma cells to temozolomide (TMZ) in vitro. Our results demonstrate that the more-responsive cell cultures displayed the longest mean fluorescence lifetime τm after TMZ treatment due to an increase in the protein-bound NAD(P)H fraction α<sub>2</sub> associated with a shift to oxidative phosphorylation. The cell cultures that responded poorly to TMZ had generally shorter τm, i.e., were more glycolytic, and showed no or insignificant changes after treatment. The FLIM data correlate well with standard measurements of cellular drug response—cell viability and proliferation index and clinical response in patients. Therefore, FLIM of NAD(P)H provides a highly sensitive, label-free assay of treatment response directly on patient-derived glioblastoma cells and can become an innovative platform for individual drug screening for patients. |
| first_indexed | 2024-03-11T02:03:25Z |
| format | Article |
| id | doaj.art-9e41e8b208b540a188c1dda57406942d |
| institution | Directory Open Access Journal |
| issn | 1424-8247 |
| language | English |
| last_indexed | 2024-03-11T02:03:25Z |
| publishDate | 2023-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceuticals |
| spelling | doaj.art-9e41e8b208b540a188c1dda57406942d2023-11-18T12:01:37ZengMDPI AGPharmaceuticals1424-82472023-05-0116679610.3390/ph16060796Measurement of Patient-Derived Glioblastoma Cell Response to Temozolomide Using Fluorescence Lifetime Imaging of NAD(P)HDiana V. Yuzhakova0Daria A. Sachkova1Marina V. Shirmanova2Artem M. Mozherov3Anna V. Izosimova4Anna S. Zolotova5Konstantin S. Yashin6Institute of Experimental Oncology and Biomedical Technologies, Privolzhsky Research Medical University, 10/1 Minin and Pozharsky Sq., 603005 Nizhny Novgorod, RussiaInstitute of Experimental Oncology and Biomedical Technologies, Privolzhsky Research Medical University, 10/1 Minin and Pozharsky Sq., 603005 Nizhny Novgorod, RussiaInstitute of Experimental Oncology and Biomedical Technologies, Privolzhsky Research Medical University, 10/1 Minin and Pozharsky Sq., 603005 Nizhny Novgorod, RussiaInstitute of Experimental Oncology and Biomedical Technologies, Privolzhsky Research Medical University, 10/1 Minin and Pozharsky Sq., 603005 Nizhny Novgorod, RussiaInstitute of Experimental Oncology and Biomedical Technologies, Privolzhsky Research Medical University, 10/1 Minin and Pozharsky Sq., 603005 Nizhny Novgorod, RussiaDepartment of Neurosurgery, Privolzhsky Research Medical University, 10/1 Minin and Pozharsky Sq., 603005 Nizhny Novgorod, RussiaDepartment of Neurosurgery, Privolzhsky Research Medical University, 10/1 Minin and Pozharsky Sq., 603005 Nizhny Novgorod, RussiaPersonalized strategies in glioblastoma treatment are highly necessary. One of the possible approaches is drug screening using patient-derived tumor cells. However, this requires reliable methods for assessment of the response of tumor cells to treatment. Fluorescence lifetime imaging microscopy (FLIM) is a promising instrument to detect early cellular response to chemotherapy using the autofluorescence of metabolic cofactors. Here, we explored FLIM of NAD(P)H to evaluate the sensitivity of patient-derived glioma cells to temozolomide (TMZ) in vitro. Our results demonstrate that the more-responsive cell cultures displayed the longest mean fluorescence lifetime τm after TMZ treatment due to an increase in the protein-bound NAD(P)H fraction α<sub>2</sub> associated with a shift to oxidative phosphorylation. The cell cultures that responded poorly to TMZ had generally shorter τm, i.e., were more glycolytic, and showed no or insignificant changes after treatment. The FLIM data correlate well with standard measurements of cellular drug response—cell viability and proliferation index and clinical response in patients. Therefore, FLIM of NAD(P)H provides a highly sensitive, label-free assay of treatment response directly on patient-derived glioblastoma cells and can become an innovative platform for individual drug screening for patients.https://www.mdpi.com/1424-8247/16/6/796glioblastomapersonalized therapypatient-derived cell culturefluorescence lifetime imaging FLIMmetabolismNAD(P)H |
| spellingShingle | Diana V. Yuzhakova Daria A. Sachkova Marina V. Shirmanova Artem M. Mozherov Anna V. Izosimova Anna S. Zolotova Konstantin S. Yashin Measurement of Patient-Derived Glioblastoma Cell Response to Temozolomide Using Fluorescence Lifetime Imaging of NAD(P)H Pharmaceuticals glioblastoma personalized therapy patient-derived cell culture fluorescence lifetime imaging FLIM metabolism NAD(P)H |
| title | Measurement of Patient-Derived Glioblastoma Cell Response to Temozolomide Using Fluorescence Lifetime Imaging of NAD(P)H |
| title_full | Measurement of Patient-Derived Glioblastoma Cell Response to Temozolomide Using Fluorescence Lifetime Imaging of NAD(P)H |
| title_fullStr | Measurement of Patient-Derived Glioblastoma Cell Response to Temozolomide Using Fluorescence Lifetime Imaging of NAD(P)H |
| title_full_unstemmed | Measurement of Patient-Derived Glioblastoma Cell Response to Temozolomide Using Fluorescence Lifetime Imaging of NAD(P)H |
| title_short | Measurement of Patient-Derived Glioblastoma Cell Response to Temozolomide Using Fluorescence Lifetime Imaging of NAD(P)H |
| title_sort | measurement of patient derived glioblastoma cell response to temozolomide using fluorescence lifetime imaging of nad p h |
| topic | glioblastoma personalized therapy patient-derived cell culture fluorescence lifetime imaging FLIM metabolism NAD(P)H |
| url | https://www.mdpi.com/1424-8247/16/6/796 |
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