JAK2V617F-positive extracellular genomic materials drive endothelial inflammation and thrombosis in myeloproliferative neoplasms

Endothelial cells (ECs) play a pivotal role in vascular inflammation and thrombosis, central processes in myeloproliferative neoplasms (MPNs). Vascular complications, particularly thrombosis, contribute significantly to MPN morbidity and mortality; however, the molecular mechanisms underlying endothelial dysfunction remain incompletely understood.

In this study, we investigated the effects of JAK2V617F-positive extracellular genomic materials (EGMs)—including cell-free DNA (cfDNA), and exosomes, derived from HEL cells—on endothelial inflammation, coagulation, and nucleic acid-sensing pathways to elucidate their contribution to MPN-associated vascular pathology. Human umbilical vein endothelial cells (HUVECs) were treated with JAK2V617F-positive EGMs and culture supernatant (CS). We assessed proliferation, apoptosis, HMGB1 expression and release, cytokine secretion, nucleic acid-sensing pathway activation, and coagulation-related gene expression via RT-PCR, ELISA, and flow cytometry.

We have demonstrated that exosomes exhibited the most potent effects on endothelial activation, robust HMGB1 secretion and cytokine release, including IL-1β, IL-6, and MCP-1, indicating activation of early inflammation while cfDNA induced intracellular HMGB1 accumulation with limited extracellular release. We further demonstrate that JAK2V617F-positive EGMs activate nucleic acid sensing pathways (cGAS-STING and TLR9), leading to increased expression of proinflammatory mediators such as NF-κB, IRF3, and IRF7. The upregulation of coagulation-related genes (F10, F3, F2RL1) and adhesion molecules (ICAM-1, TF) confirmed that EGM exposure promotes a pro-thrombotic phenotype in ECs, mirroring the hypercoagulable state in MPN patients.

As our in vitro system lacks physiological conditions such as blood flow dynamics and immune cell interactions, further validation in patient-derived samples and in vivo models with varying JAK2V617F allele burdens is required. Nonetheless, these findings establish a mechanistic link between oncogenic EGMs and vascular complications in MPNs, highlighting their role in chronic inflammation and thrombosis. Targeting EGM-mediated endothelial dysfunction may offer a novel therapeutic approach to mitigate vascular complications in MPN patients.