Clinically Relevant Oxygraphic Assay to Assess Mitochondrial Energy Metabolism in Acute Myeloid Leukemia Patients
Resistant acute myeloid leukemia (AML) exhibits mitochondrial energy metabolism changes compared to newly diagnosed AML. This phenotype is often observed by evaluating the mitochondrial oxygen consumption of blasts, but most of the oximetry protocols were established from leukemia cell lines without...
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MDPI AG
2021-12-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/13/24/6353 |
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| author | Quentin Fovez William Laine Laure Goursaud Celine Berthon Nicolas Germain Claire Degand Jean-Emmanuel Sarry Bruno Quesnel Philippe Marchetti Jerome Kluza |
| author_facet | Quentin Fovez William Laine Laure Goursaud Celine Berthon Nicolas Germain Claire Degand Jean-Emmanuel Sarry Bruno Quesnel Philippe Marchetti Jerome Kluza |
| author_sort | Quentin Fovez |
| collection | DOAJ |
| description | Resistant acute myeloid leukemia (AML) exhibits mitochondrial energy metabolism changes compared to newly diagnosed AML. This phenotype is often observed by evaluating the mitochondrial oxygen consumption of blasts, but most of the oximetry protocols were established from leukemia cell lines without validation on primary leukemia cells. Moreover, the cultures and storage conditions of blasts freshly extracted from patient blood or bone marrow cause stress, which must be evaluated before determining oxidative phosphorylation (OXPHOS). Herein, we evaluated different conditions to measure the oxygen consumption of blasts using extracellular flow analyzers. We first determined the minimum number of blasts required to measure OXPHOS. Next, we compared the OXPHOS of blasts cultured for 3 h and 18 h after collection and found that to maintain metabolic organization for 18 h, cytokine supplementation is necessary. Cytokines are also needed when measuring OXPHOS in cryopreserved, thawed and recultured blasts. Next, the concentrations of respiratory chain inhibitors and uncoupler FCCP were established. We found that the FCCP concentration required to reach the maximal respiration of blasts varied depending on the patient sample analyzed. These protocols provided can be used in future clinical studies to evaluate OXPHOS as a biomarker and assess the efficacy of treatments targeting mitochondria. |
| first_indexed | 2024-03-10T04:32:04Z |
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| id | doaj.art-b6b0443d40f84d48bd1fc6b86bf833e2 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-10T04:32:04Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
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| series | Cancers |
| spelling | doaj.art-b6b0443d40f84d48bd1fc6b86bf833e22023-11-23T04:07:33ZengMDPI AGCancers2072-66942021-12-011324635310.3390/cancers13246353Clinically Relevant Oxygraphic Assay to Assess Mitochondrial Energy Metabolism in Acute Myeloid Leukemia PatientsQuentin Fovez0William Laine1Laure Goursaud2Celine Berthon3Nicolas Germain4Claire Degand5Jean-Emmanuel Sarry6Bruno Quesnel7Philippe Marchetti8Jerome Kluza9Institut pour la Recherche sur le Cancer de Lille, Univ. Lille, CNRS, Inserm, CHU Lille, UMR9020-UMR-S 1277-Canther-Cancer Heterogeneity, Plasticity and Resistance to Therapies, F-59000 Lille, FranceInstitut pour la Recherche sur le Cancer de Lille, Univ. Lille, CNRS, Inserm, CHU Lille, UMR9020-UMR-S 1277-Canther-Cancer Heterogeneity, Plasticity and Resistance to Therapies, F-59000 Lille, FranceInstitut pour la Recherche sur le Cancer de Lille, Univ. Lille, CNRS, Inserm, CHU Lille, UMR9020-UMR-S 1277-Canther-Cancer Heterogeneity, Plasticity and Resistance to Therapies, F-59000 Lille, FranceHematology Department, CHU Lille, F-59000 Lille, FranceInstitut pour la Recherche sur le Cancer de Lille, Univ. Lille, CNRS, Inserm, CHU Lille, UMR9020-UMR-S 1277-Canther-Cancer Heterogeneity, Plasticity and Resistance to Therapies, F-59000 Lille, FranceInstitut pour la Recherche sur le Cancer de Lille, Univ. Lille, CNRS, Inserm, CHU Lille, UMR9020-UMR-S 1277-Canther-Cancer Heterogeneity, Plasticity and Resistance to Therapies, F-59000 Lille, FranceCentre National de la Recherche Scientifique, Centre de Recherches en Cancérologie de Toulouse, Institut National de la Santé et de la Recherche Médicale, Université de Toulouse, 31100 Toulouse, FranceInstitut pour la Recherche sur le Cancer de Lille, Univ. Lille, CNRS, Inserm, CHU Lille, UMR9020-UMR-S 1277-Canther-Cancer Heterogeneity, Plasticity and Resistance to Therapies, F-59000 Lille, FranceInstitut pour la Recherche sur le Cancer de Lille, Univ. Lille, CNRS, Inserm, CHU Lille, UMR9020-UMR-S 1277-Canther-Cancer Heterogeneity, Plasticity and Resistance to Therapies, F-59000 Lille, FranceInstitut pour la Recherche sur le Cancer de Lille, Univ. Lille, CNRS, Inserm, CHU Lille, UMR9020-UMR-S 1277-Canther-Cancer Heterogeneity, Plasticity and Resistance to Therapies, F-59000 Lille, FranceResistant acute myeloid leukemia (AML) exhibits mitochondrial energy metabolism changes compared to newly diagnosed AML. This phenotype is often observed by evaluating the mitochondrial oxygen consumption of blasts, but most of the oximetry protocols were established from leukemia cell lines without validation on primary leukemia cells. Moreover, the cultures and storage conditions of blasts freshly extracted from patient blood or bone marrow cause stress, which must be evaluated before determining oxidative phosphorylation (OXPHOS). Herein, we evaluated different conditions to measure the oxygen consumption of blasts using extracellular flow analyzers. We first determined the minimum number of blasts required to measure OXPHOS. Next, we compared the OXPHOS of blasts cultured for 3 h and 18 h after collection and found that to maintain metabolic organization for 18 h, cytokine supplementation is necessary. Cytokines are also needed when measuring OXPHOS in cryopreserved, thawed and recultured blasts. Next, the concentrations of respiratory chain inhibitors and uncoupler FCCP were established. We found that the FCCP concentration required to reach the maximal respiration of blasts varied depending on the patient sample analyzed. These protocols provided can be used in future clinical studies to evaluate OXPHOS as a biomarker and assess the efficacy of treatments targeting mitochondria.https://www.mdpi.com/2072-6694/13/24/6353energy metabolismleukemiaresistancefunctional biomarkeruncoupling respirationOCR |
| spellingShingle | Quentin Fovez William Laine Laure Goursaud Celine Berthon Nicolas Germain Claire Degand Jean-Emmanuel Sarry Bruno Quesnel Philippe Marchetti Jerome Kluza Clinically Relevant Oxygraphic Assay to Assess Mitochondrial Energy Metabolism in Acute Myeloid Leukemia Patients Cancers energy metabolism leukemia resistance functional biomarker uncoupling respiration OCR |
| title | Clinically Relevant Oxygraphic Assay to Assess Mitochondrial Energy Metabolism in Acute Myeloid Leukemia Patients |
| title_full | Clinically Relevant Oxygraphic Assay to Assess Mitochondrial Energy Metabolism in Acute Myeloid Leukemia Patients |
| title_fullStr | Clinically Relevant Oxygraphic Assay to Assess Mitochondrial Energy Metabolism in Acute Myeloid Leukemia Patients |
| title_full_unstemmed | Clinically Relevant Oxygraphic Assay to Assess Mitochondrial Energy Metabolism in Acute Myeloid Leukemia Patients |
| title_short | Clinically Relevant Oxygraphic Assay to Assess Mitochondrial Energy Metabolism in Acute Myeloid Leukemia Patients |
| title_sort | clinically relevant oxygraphic assay to assess mitochondrial energy metabolism in acute myeloid leukemia patients |
| topic | energy metabolism leukemia resistance functional biomarker uncoupling respiration OCR |
| url | https://www.mdpi.com/2072-6694/13/24/6353 |
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