Neutrophil-specific expression of JAK2-V617F or CALRmut induces distinct inflammatory profiles in myeloproliferative neoplasia

• Verfasst von: Haage, Tobias Ronny [VerfasserIn]
• Verfasst von: Charakopoulos, Emmanouil [VerfasserIn]
• Verfasst von: Bhuria, Vikas [VerfasserIn]
• Verfasst von: Baldauf, Conny K. [VerfasserIn]
• Verfasst von: Korthals, Mark [VerfasserIn]
• Verfasst von: Handschuh, Juliane [VerfasserIn]
• Verfasst von: Müller, Peter [VerfasserIn]
• Verfasst von: Li, Juan [VerfasserIn]
• Verfasst von: Harit, Kunjan [VerfasserIn]
• Verfasst von: Nishanth, Gopala [VerfasserIn]
• Verfasst von: Frey, Stephanie [VerfasserIn]
• Verfasst von: Böttcher, Martin [VerfasserIn]
• Verfasst von: Fischer, Klaus-Dieter [VerfasserIn]
• Verfasst von: Dudeck, Jan [VerfasserIn]
• Verfasst von: Dudeck, Anne [VerfasserIn]
• Verfasst von: Lipka, Daniel [VerfasserIn]
• Verfasst von: Schraven, Burkhart [VerfasserIn]
• Verfasst von: Green, Tony [VerfasserIn]
• Verfasst von: Müller, Andreas Johann [VerfasserIn]
• Verfasst von: Mougiakakos, Dimitrios [VerfasserIn]
• Verfasst von: Fischer, Thomas [VerfasserIn]
• Titel: Neutrophil-specific expression of JAK2-V617F or CALRmut induces distinct inflammatory profiles in myeloproliferative neoplasia
• Verf.angabe: Tobias Ronny Haage, Emmanouil Charakopoulos, Vikas Bhuria, Conny K. Baldauf, Mark Korthals, Juliane Handschuh, Peter Müller, Juan Li, Kunjan Harit, Gopala Nishanth, Stephanie Frey, Martin Böttcher, Klaus-Dieter Fischer, Jan Dudeck, Anne Dudeck, Daniel B. Lipka, Burkhart Schraven, Anthony R. Green, Andreas J. Müller, Dimitrios Mougiakakos and Thomas Fischer
• E-Jahr: 2024
• Jahr: 09 June 2024
• Umfang: 23 S.
• Illustrationen: Illustrationen
• Fussnoten: Gesehen am 18.11.2024
• Titel Quelle: Enthalten in: Journal of hematology & oncology
• Ort Quelle: London : Biomed Central, 2008
• Jahr Quelle: 2024
• Band/Heft Quelle: 17(2024), Artikel-ID 43, Seite 1-23
• ISSN Quelle: 1756-8722
• DOI: doi:10.1186/s13045-024-01562-5
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1907996907

Abstract

Background: Neutrophils play a crucial role in inflammation and in the increased thrombotic risk in myeloproliferative neoplasms (MPNs). We have investigated how neutrophil-specific expression of JAK2-V617F or CALRdel re-programs the functions of neutrophils. Methods: Ly6G-Cre JAK2-V617F and Ly6G-Cre CALRdel mice were generated. MPN parameters as blood counts, splenomegaly and bone marrow histology were compared to wild-type mice. Megakaryocyte differentiation was investigated using lineage-negative bone marrow cells upon in vitro incubation with TPO/IL-1β. Cytokine concentrations in serum of mice were determined by Mouse Cytokine Array. IL-1α expression in various hematopoietic cell populations was determined by intracellular FACS analysis. RNA-seq to analyse gene expression of inflammatory cytokines was performed in isolated neutrophils from JAK2-V617F and CALR-mutated mice and patients. Bioenergetics of neutrophils were recorded on a Seahorse extracellular flux analyzer. Cell motility of neutrophils was monitored in vitro (time lapse microscopy), and in vivo (two-photon microscopy) upon creating an inflammatory environment. Cell adhesion to integrins, E-selectin and P-selection was investigated in-vitro. Statistical analysis was carried out using GraphPad Prism. Data are shown as mean ± SEM. Unpaired, two-tailed t-tests were applied. Results: Strikingly, neutrophil-specific expression of JAK2-V617F, but not CALRdel, was sufficient to induce pro-inflammatory cytokines including IL-1 in serum of mice. RNA-seq analysis in neutrophils from JAK2-V617F mice and patients revealed a distinct inflammatory chemokine signature which was not expressed in CALR-mutant neutrophils. In addition, IL-1 response genes were significantly enriched in neutrophils of JAK2-V617F patients as compared to CALR-mutant patients. Thus, JAK2-V617F positive neutrophils, but not CALR-mutant neutrophils, are pathogenic drivers of inflammation in MPN. In line with this, expression of JAK2-V617F or CALRdel elicited a significant difference in the metabolic phenotype of neutrophils, suggesting a stronger inflammatory activity of JAK2-V617F cells. Furthermore, JAK2-V617F, but not CALRdel, induced a VLA4 integrin-mediated adhesive phenotype in neutrophils. This resulted in reduced neutrophil migration in vitro and in an inflamed vessel. This mechanism may contribute to the increased thrombotic risk of JAK2-V617F patients compared to CALR-mutant individuals. Conclusions: Taken together, our findings highlight genotype-specific differences in MPN-neutrophils that have implications for the differential pathophysiology of JAK2-V617F versus CALR-mutant disease.