Glucose transport dependency defines a therapeutic vulnerability in JAK2V617F-driven myeloproliferative neoplasms

Myeloproliferative neoplasms (MPN) comprise a heterogenous group of hematological malignancies that include polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). Current therapeutic strategies rely on cytoreductive approaches that mitigate disease burden and thromboembolic risk but are not curative. Allogeneic stem cell transplantation remains the only curative option, underscoring the need for novel therapeutic strategies. We previously identified hypoxia-inducible factor 1 (HIF-‍‍1) as a selective vulnerability in JAK2V617F-positive cells, but the underlying metabolic mechanisms remain incompletely defined.

In vitro studies utilized 32D cells transduced with an empty vector control, Jak2WT, or Jak2V617F. To evaluate metabolic dependencies, CRISPR-Cas9 was used to generate Slc2a1 (GLUT1) and Slc2a3 (GLUT3) knockout clones, which were subsequently characterized via RNA sequencing, extracellular flux analysis, and cellular fitness assays (proliferation, viability, and apoptosis). Pharmacological targeted inhibition of GLUT1/3 was evaluated in human JAK2V617F-mutated post-MPN AML cell lines (SET-2, HEL), primary patient-derived cells and a Jak2V617F knock-in mouse model. Combinatorial efficacy was assessed using the JAK1/2 inhibitor ruxolitinib.

JAK2V617F induced HIF-1-dependent metabolic reprogramming, characterized by increased glycolytic flux and oxidative metabolism. Complete abrogation of glucose uptake occurred only upon combined loss of GLUT1 and GLUT3 in Jak2V617F cells, revealing functional redundancy between these transporters that sustains enhanced glycolysis. Disruption of glucose uptake selectively induced stress-associated transcriptional programs and replication stress, triggering an S-phase arrest that culminated in apoptosis and impaired viability, specifically in Jak2V617F cells. In vivo, pharmacological inhibition of HIF-1 or GLUT induced a reorganization of erythropoiesis to the spleen but did not ameliorate core disease features. In contrast, in vitro GLUT inhibition robustly reduced cell viability in human SET-2 and HEL cell lines and impaired proliferation, viability, and colony formation in patient-derived PBMCs.

Collectively, these findings establish HIF-1-driven glucose metabolism as a metabolic vulnerability in JAK2V617F-positive MPN. The selective exhaustion of patient-derived clones defines the HIF-1-GLUT1/3 axis as a central, targetable bottleneck. These data provide a mechanistic rationale for further investigation of HIF-1 or GLUT inhibitors, suggesting that targeting this fundamental requirement may help overcome clinical limitations to achieve disease modification and eradicate the malignant clone.