A research team led by Dr. Josep Francesc Nomdedeu, head of the Hematologic Diagnostics Research Group at the Sant Pau Research Institute (IR Sant Pau) and hematologist at the Sant Pau Hospital’s Hematology Laboratory, in collaboration with the Universitat Autònoma de Barcelona, the University of Barcelona, the Cancer Research Center of Marseille (INSERM), and other institutions, has identified a previously underexplored mechanism by which the KIT D816V mutation reprograms cellular metabolism to promote tumor development.
The study, recently published in the journal Experimental Hematology, shows that the KIT D816V mutation—frequently found in hematologic malignancies such as acute myeloid leukemia (AML) and systemic mastocytosis—increases both the number and activity of mitochondria while simultaneously reducing their physiological clearance through mitophagy. Mitophagy is a selective cellular cleanup process through which damaged or unneeded mitochondria are eliminated to maintain metabolic balance and prevent the buildup of harmful free radicals. The central mechanism behind this disruption involves the downregulation of the BNIP3 gene, a key regulator of mitochondrial autophagy.
To investigate the impact of the KIT D816V mutation, researchers used two human myeloid cell lines: ROSA (mast cell-like) and TF-1 (erythroleukemia). They compared the behavior of mutated cells to their wild-type (non-mutated) counterparts.
The team analyzed various biochemical and cellular parameters to assess mitochondrial function, including oxygen consumption, ATP production, mitochondrial membrane potential, and reactive oxygen species (ROS) generation. They also measured the number, size, and morphology of mitochondria, along with the expression of critical markers related to autophagy and mitophagy—such as BNIP3 and LC3—using electron microscopy, flow cytometry, qPCR, and immunoblotting.
To confirm the clinical relevance of these findings, researchers analyzed samples from AML patients to determine whether those with the KIT D816V mutation also showed reduced BNIP3 levels. Indeed, this association was observed.
Unlike many other cancers that rely primarily on glycolysis—a fast but less efficient metabolic process that generates energy from glucose without requiring oxygen—this study found that KIT D816V-mutant cells favor mitochondrial oxidative phosphorylation (OXPHOS) to produce energy. This shift results in increased ATP production, heightened mitochondrial respiration, and elevated levels of ROS such as superoxide, which can damage DNA and contribute to genomic instability.
The researchers found that this functional shift is directly linked to lower levels of BNIP3, a protein that serves as a signal for the selective degradation of defective mitochondria. Its loss leads to an accumulation of mitochondria that are active but dysfunctional, potentially increasing oxidative stress and promoting tumor progression.
In patient-derived AML samples, those harboring the KIT D816V mutation exhibited significantly lower BNIP3 levels, which were associated with worse overall survival.
The study also revealed that the mTOR pathway—one of the primary regulators of autophagy—is constitutively activated in mutant cells, even under nutrient-deprived or growth factor-deficient conditions. This autonomous function reflects a loss of dependence on external signals, a hallmark of many cancer cells.
“The KIT D816V mutation acts as a double accelerator: it boosts cellular energy production through mitochondria while also blocking their natural elimination,” said Dr. Josep F. Nomdedeu. “This imbalance could pave the way for new therapeutic strategies aimed at restoring control over mitochondrial function.”
The findings suggest that restoring BNIP3 expression could be a viable strategy to counteract the pro-tumor effects of the KIT mutation. In in vitro experiments, overexpression of BNIP3 reinstated mitophagy and reduced the number of mitochondria in mutant cells.
This discovery opens new avenues for research and potential therapeutic targets in hematologic malignancies driven by KIT mutations, particularly in patients who are resistant to conventional treatments.
This study was supported by multiple national and international research institutions. In Spain, funding came from the Carlos III Health Institute through projects PI13/2729 and PI16/094, and from the Generalitat de Catalunya via grants 2014-SGR-383, 2017-SGR-1395, and the PERIS program SLT 002/16/0043.
Internationally, the research was funded by Aix-Marseille University, the Amidex Foundation, Canceropôle Provence-Alpes-Côte d’Azur, the French National Cancer Institute (INCa), and the Provence-Alpes-Côte d’Azur Region, highlighting the collaborative and translational nature of the project.
Cisa-Wieczorek S, Hernández-Alvarez MI, Parreño M, Muñoz JP, Bussaglia E, Carricondo M, Ubeda J, Dubreuil P, Zorzano A, Brenet F, Nomdedeu JF. D816V KIT mutation induces mitochondrial morphologic and functional changes through BNIP3 downregulation in human myeloid cell lines ROSA and TF-1. Exp Hematol. 2025;145:104748. https://doi.org/10.1016/j.exphem.2025.104748
