In groundbreaking research published in Nature Metabolism, scientists have uncovered a previously unrecognized metabolic vulnerability in acute myeloid leukemia (AML) that centers on the critical role of mitochondrial complex II in purine biosynthesis. This discovery emerges from a comprehensive pathway coessentiality mapping approach, revealing that the inhibition of complex II triggers an accumulation of glutamate (Glu) upstream of the enzyme, fundamentally disrupting cellular metabolism and impairing leukemia cell growth and survival.
Mitochondrial complex II, also known as succinate dehydrogenase, is well-known for its dual role in the tricarboxylic acid (TCA) cycle and the electron transport chain, making it essential for energy production in cells. In AML, researchers have discerned that blockage of complex II leads to a significant build-up of critical metabolites including succinate, alpha-ketoglutarate (α-KG), and glutamate, with profound downstream effects on amino acid pathways. The intricate metabolic crosstalk exposed by these findings sheds light on how leukemic cells heavily rely on functional complex II to maintain their proliferative and survival advantages.
To probe the biological impact of metabolite accumulation, the research team utilized cell-permeable dimethyl esters: dimethyl-succinate (DMS), dimethyl-alpha-KG (DMKG), and dimethyl-glutamate (DMG). Remarkably, glutamate accumulation, simulated by DMG treatment, exhibited a cytotoxic effect tenfold more potent than DMS or DMKG across AML cells harboring diverse genetic mutations such as KMT2A, FLT3, NRAS, and TP53. This striking sensitivity to DMG underscores glutamate’s pivotal role as a metabolic node that, when dysregulated, can collapse leukemic cell viability.
Delving deeper, the study revealed that AML cells react to complex II inhibition by rerouting metabolic fluxes. Glutamate produced during purine biosynthesis, wherein glutamine donates nitrogen to key intermediates, normally supports the TCA cycle via anaplerosis—refilling cycle intermediates that sustain energy and biosynthetic processes. However, upon complex II blockade, glutamate fails to effectively enter the TCA cycle, leading to its intracellular accumulation. This accumulation not only inhibits purine synthesis but also entails glutamate export through system Xc− and incorporation into glutathione biosynthesis, processes intimately linked to redox homeostasis.
The ramifications of glutamate accumulation were strikingly confirmed by metabolic assays showing that three distinct complex II inhibitors increased intracellular glutamate levels in AML lines. Interestingly, the addition of exogenous glutamate heightened cell sensitivity to complex II inhibition, suggesting that glutamate overload exacerbates metabolic stress and compromises leukemia cell fitness. This interrelationship between glutamate handling and complex II function reveals a metabolic bottleneck exploitable for therapeutic intervention.
A significant insight from this research is the inability of glutamate to compensate for glutamine withdrawal in AML. Normally, glutamine-derived carbons replenish the TCA cycle, but the study’s results highlight that dimethyl-succinate and dimethyl-alpha-KG can rescue AML cell growth under low-glutamine conditions, unlike dimethyl-glutamate. This differential effect cements the concept that glutamine metabolism in AML depends heavily on downstream metabolites that feed the TCA cycle rather than glutamate per se, which may have toxic secondary effects when accumulated.
The role of glutamate in regulating redox balance emerged as a crucial adaptive pathway in the face of complex II inhibition. Glutamate is a precursor for glutathione synthesis and modulates cystine import via system Xc−, critical for maintaining antioxidant defenses. The research demonstrated that complex II blockade induced significant changes in glutathione metabolism, pushing AML cells to channel glutamate toward glutathione production. Inhibition of system Xc− with erastin further elevated intracellular glutamate and diminished cell viability synergistically with complex II inhibitors, illustrating the therapeutic potential of co-targeting glutamate metabolism.
Metabolomic profiling unveiled that glutamate accumulation following DMG treatment profoundly suppresses purine nucleotide pools, including key intermediates such as 5-amino-4-imidazolecarboxamide ribonucleotide (AICAR), adenylosuccinate, ADP, and ATP. By downregulating purine biosynthesis, glutamate accumulation undermines nucleotide availability essential for DNA and RNA synthesis, thereby hampering AML cell proliferation and survival. This mechanistic understanding elucidates how complex II functions not only as a metabolic enzyme but also as an indirect regulator of nucleotide biosynthesis essential for leukemia maintenance.
The discovery of this metabolic chokepoint offers a promising new avenue for AML therapy. Targeting complex II to induce glutamate accumulation, possibly in combination with agents that limit glutamate export or glutathione biosynthesis, could synergistically amplify metabolic stress and selectivity against leukemic cells. This strategy exploits a fundamental biochemical dependency of AML cells on complex II integrity and glutamate homeostasis, representing a precision medicine approach grounded in metabolic reprogramming.
Notably, the study underscores that AML’s metabolic wiring differs markedly from normal hematopoietic cells, which may explain the unique susceptibility of leukemia to complex II inhibition and glutamate toxicity. By pinpointing glutamate as a key mediator of purine biosynthesis inhibition, this research challenges the traditional perspective that mitochondrial defects simply reduce energy generation, instead emphasizing their nuanced role in shaping nucleotide metabolism and redox balance in cancer.
Importantly, the implication that glutamate accumulation acts as a metabolic stress signal, suppressing purine synthesis, situates this metabolite as more than a passive substrate but as an active regulator of cellular biosynthetic capacity. This insight prompts reconsideration of metabolic intermediates as dynamic regulators in cancer biology with roles extending beyond their canonical biochemical functions.
The elucidation of this pathway also raises intriguing questions about the broader applicability of complex II targeting beyond AML. Given that many cancers display altered mitochondrial metabolism and nucleotide synthesis demands, the mechanism described may represent a generalized vulnerability. However, the specificity seen in AML suggests that genetic and metabolic context dictate responses to complex II inhibition and glutamate-induced stress.
Furthermore, the study’s methodological approach, harnessing pathway coessentiality mapping combined with isotope tracing and metabolic assays, establishes a robust framework for unraveling complex biochemical networks in cancer. This multifaceted strategy provides comprehensive insights linking enzyme function, metabolite fluxes, and cellular phenotypes, advancing our understanding of metabolic dependencies in malignancy.
Overall, this research shifts the paradigm in leukemia metabolism by revealing a critical dependency on complex II for maintaining purine biosynthesis via glutamate regulation. The findings spotlight new metabolic checkpoints amenable to therapeutic intervention and propose combinatorial strategies that enhance glutamate-induced cytotoxicity alongside mitochondrial inhibition. As such, this study paves the way for innovative metabolic therapies combatting AML with unprecedented precision and efficacy.
In conclusion, the identification of glutamate accumulation as a metabolic liability in AML upon complex II inhibition represents a major stride in cancer metabolism research. This work not only elucidates novel biochemical pathways underpinning leukemia survival but also inspires new therapeutic directions that exploit metabolic vulnerabilities to eradicate malignant cells. The promise of targeting these intertwined metabolic axes holds significant potential to improve outcomes for patients afflicted with this aggressive hematologic cancer.
Subject of Research:
Acute myeloid leukemia (AML) metabolism; role of mitochondrial complex II in purine biosynthesis and glutamate accumulation.
Article Title:
Pathway coessentiality mapping reveals complex II is required for de novo purine biosynthesis in acute myeloid leukaemia.
Article References:
Stewart, A.E., Zachman, D.K., Castellano-Escuder, P. et al. Pathway coessentiality mapping reveals complex II is required for de novo purine biosynthesis in acute myeloid leukaemia. Nat Metab (2025). https://doi.org/10.1038/s42255-025-01410-x
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AI Generated
DOI:
https://doi.org/10.1038/s42255-025-01410-x
Tags: acute myeloid leukemiaamino acid metabolic pathwayscellular metabolism disruptioncytotoxic effects of metabolitesdimethyl esters in cancer researchglutamate accumulation effectsleukemia cell survival mechanismsmetabolic vulnerability in leukemiamitochondrial complex II rolepurine biosynthesis in AMLsuccinate dehydrogenase functionTCA cycle and leukemia
