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31P and 13C NMR studies of isolated perfused hematopoietic cells from leukemic mice
F Mégnin1, J F Nedelec, J L Dimicoli
1Institut Curie, Biologie, (INSERM U.219), Centre Universitaire, Orsay, France.
Abstract:
The myeloproliferative leukemic virus (MPLV) induces within 2-3 weeks a massive infiltration of the adult mouse liver by hematopoietic leukemic cells. Since the metabolism of the infiltrated organ might be correlated with an interaction of two cell populations, it was decided to study the isolated hematopoietic cells separately. The metabolism of these cells embedded in an agarose gel and perfused with labeled substrates was investigated using 31P and 13C NMR. Using [1-13C]glucose as precursor, sequential 13C NMR spectra showed that the hematopoietic cells were able to store glucose as [1-13C]glycogen and to metabolize it through the glycolytic pathway to give [3-13C]lactate as sole end-product. The liver neoglucogenic substrates: [2-13C]pyruvate and [3-13C]alanine are not metabolized by these cells. This suggests that the tricarboxylic acid cycle was not efficient. To investigate further the glycolytic properties of the cells, 10 mM sodium azide was added to the medium containing [1-13C]glucose. When compared to the aerobic conditions, a 40% decrease of nucleotides (0.10 vs 0.17 mumole NTP/10(9) cells), a degradation of [1-13C]glycogen and an increase of ca 35% of the glycolytic rate were observed. The analysis of 13C NMR spectra of the perfusates at the end of the perfusion indicates a total conversion of [1-13C]glucose into [3-13C]lactate and [3-13C]pyruvate under anaerobic conditions. These results permit a better understanding of the metabolism of the perfused leukemic livers which are extensively infiltrated by these hematopoietic cells.
Insights
Myeloproliferative leukemic virus (MPLV) causes leukemic cell infiltration in mouse livers. Isolated leukemic cells primarily metabolize glucose to lactate, indicating inefficient energy pathways and altered nucleotide levels under anaerobic conditions.
Area of Science:
- Biochemistry
- Cell Biology
- Oncology
Background:
- Myeloproliferative leukemic virus (MPLV) rapidly infiltrates adult mouse livers with hematopoietic leukemic cells.
- Understanding leukemic cell metabolism is crucial for comprehending organ infiltration and function.
- Investigating isolated leukemic cell metabolism provides insights into their specific metabolic activities.
Purpose of the Study:
- To investigate the metabolic pathways of hematopoietic leukemic cells isolated from MPLV-infiltrated mouse livers.
- To determine the metabolic fate of glucose and other substrates in these leukemic cells.
- To assess the impact of anaerobic conditions on leukemic cell metabolism.
Main Methods:
- Utilized 31P and 13C Nuclear Magnetic Resonance (NMR) spectroscopy.
- Studied isolated leukemic cells embedded in agarose gel and perfused with labeled substrates ([1-13C]glucose, [2-13C]pyruvate, [3-13C]alanine).
- Introduced sodium azide to induce anaerobic conditions and analyzed metabolic changes.
Main Results:
- Leukemic cells stored [1-13C]glucose as [1-13C]glycogen and metabolized it to [3-13C]lactate as the sole end-product.
- [2-13C]pyruvate and [3-13C]alanine were not metabolized, suggesting an inefficient tricarboxylic acid cycle.
- Under anaerobic conditions, nucleotide levels decreased, glycogen degraded, and glycolytic rate increased, with complete conversion of glucose to lactate and pyruvate.
Conclusions:
- Leukemic cells exhibit a predominantly glycolytic metabolism, converting glucose to lactate.
- The tricarboxylic acid cycle appears inefficient in these leukemic cells.
- Metabolic alterations, particularly under anaerobic conditions, provide a deeper understanding of leukemic liver infiltration.