Abstract
Targeting altered tumor cell metabolism might provide an attractive opportunity for patients with acute myeloid leukemia (AML). An amino acid dropout screen on primary leukemic stem cells and progenitor populations revealed a number of amino acid dependencies, of which methionine was one of the strongest. By using various metabolite rescue experiments, nuclear magnetic resonance−based metabolite quantifications and 13C-tracing, polysomal profiling, and chromatin immunoprecipitation sequencing, we identified that methionine is used predominantly for protein translation and to provide methyl groups to histones via S-adenosylmethionine for epigenetic marking. H3K36me3 was consistently the most heavily impacted mark following loss of methionine. Methionine depletion also reduced total RNA levels, enhanced apoptosis, and induced a cell cycle block. Reactive oxygen species levels were not increased following methionine depletion, and replacement of methionine with glutathione or N-acetylcysteine could not rescue phenotypes, excluding a role for methionine in controlling redox balance control in AML. Although considered to be an essential amino acid, methionine can be recycled from homocysteine. We uncovered that this is primarily performed by the enzyme methionine synthase and only when methionine availability becomes limiting. In vivo, dietary methionine starvation was not only tolerated by mice, but also significantly delayed both cell line and patient-derived AML progression. Finally, we show that inhibition of the H3K36-specific methyltransferase SETD2 phenocopies much of the cytotoxic effects of methionine depletion, providing a more targeted therapeutic approach. In conclusion, we show that methionine depletion is a vulnerability in AML that can be exploited therapeutically, and we provide mechanistic insight into how cells metabolize and recycle methionine.
| Original language | English |
|---|---|
| Journal | Blood |
| Volume | 140 |
| Issue number | 19 |
| Pages (from-to) | 2037-2052 |
| Number of pages | 16 |
| ISSN | 0006-4971 |
| DOIs | |
| Publication status | Published - 10.11.2022 |
Funding
These studies were financially supported by the EU (H2020-MSCA-ITN-2015-675790-HaemMetabolome) awarded to J.J.S. and U.G. In addition, A.C., A.E., and I.A. gratefully acknowledge receipt of a Marie Curie Fellowship and are participants in the same Initial Training Network. Work at BMRZ is supported by the state of Hesse. The authors gratefully acknowledge Marcel de Vries and Hjalmar Permentier from the proteomics department of the ERIBA Institute Groningen for their help with methionine quantifications. The authors also thank Christina Muhs and Christian Richter for their help with the NMR spectroscopy measurements. The authors gratefully acknowledge Olav Rooyackers for his critical review of the manuscript. The authors are grateful to Marcel van Vugt for providing many of the cell cycle, apoptosis, and anti-puromycin antibodies. These studies were financially supported by the EU (H2020-MSCA-ITN-2015-675790-HaemMetabolome) awarded to J.J.S. and U.G. In addition, A.C. A.E. and I.A. gratefully acknowledge receipt of a Marie Curie Fellowship and are participants in the same Initial Training Network. Work at BMRZ is supported by the state of Hesse. Contribution: A.C. and A.E. led the study from the beginning, contributed to the study design, and performed many of the experiments and data analysis, and prepared the figures for the manuscript; A.C. prepared the initial draft of the manuscript; A.E. critically reviewed the manuscript and final figures; at a later stage in the project, A.C. overtook implementation of experiments; I.A. ran all NMR spectroscopy samples and performed all of the associated analysis under the supervision of H.S.; M.G. and F.A.J.v.d.H. assisted throughout with experimental work, performing many of the metabolite supplementation experiments as well as western blots; M.P. contributed to the design of the AML mouse studies, their implementation and oversaw the planning, feeding, and sacrifice of mice; A.T.J.W. assisted with the design and cloning of short-hairpin RNAs, and participated in project discussions throughout; H.t.B. performed many of the cell line methionine titrations, worked on the methionine synthase (MTR) knockdown studies, and testing the combination of methionine depletion with decitabine; R.D. assisted with the design and performance of the polysomal profiling experiments under the supervision of C.W.; D.A.P.-M. provided samples and performed reverse transcription−polymerase chain reaction analysis as well as drug/methionine depletion combinations, the healthy mouse methionine dietary starvation study, as well as Seahorse and other experiments; I.W. helped conduct the healthy mouse dietary methionine starvation study; V.v.d.B. and S.M.H. performed chromatin immunoprecipitation (ChIP) reactions and sequencing analysis; R.K. ran all DNA methylation mass spectrometry samples as well their data analysis and interpretation under the supervision and financial support of J.H.J.; P.d.B. and M.R.H.-F. performed medium metabolite quantification in MTR genetic models, as well as the associated data analysis; U.L.G. was the awardee of the grant and partially supervised A.C.; E.M.R. provided the facilities for the healthy mouse methionine starvation study and cosupervised D.A.P.-M. and I.W.; J.H.A.M. performed the sequencing on chromatin immunoprecipitation sequencing samples; G.H. cosupervised A.C. and A.E. and was involved in discussions throughout; J.J.S. was responsible for the conceptualization of the study, supervision throughout, data analysis and interpretation of data, as well as preparation of the figures and manuscript; and all authors reviewed the manuscript.
Research Areas and Centers
- Academic Focus: Center for Brain, Behavior and Metabolism (CBBM)
- Research Area: Luebeck Integrated Oncology Network (LION)
- Centers: University Cancer Center Schleswig-Holstein (UCCSH)
DFG Research Classification Scheme
- 2.22-14 Hematology, Oncology
- 2.22-17 Endocrinology, Diabetology, Metabolism
- 2.22-05 Nutritional Sciences
