Multiomic profiling reveals metabolic alterations mediating aberrant platelet activity and inflammation in myeloproliferative neoplasms
Fan He, … , Jorge Di Paola, Stephen T. Oh
Fan He, … , Jorge Di Paola, Stephen T. Oh
Published December 7, 2023
Citation Information: J Clin Invest. 2024;134(3):e172256. https://doi.org/10.1172/JCI172256.
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Research Article Hematology

Multiomic profiling reveals metabolic alterations mediating aberrant platelet activity and inflammation in myeloproliferative neoplasms

Abstract

Platelets from patients with myeloproliferative neoplasms (MPNs) exhibit a hyperreactive phenotype. Here, we found elevated P-selectin exposure and platelet-leukocyte aggregates indicating activation of platelets from essential thrombocythemia (ET) patients. Single-cell RNA-seq analysis of primary samples revealed significant enrichment of transcripts related to platelet activation, mTOR, and oxidative phosphorylation in ET patient platelets. These observations were validated via proteomic profiling. Platelet metabolomics revealed distinct metabolic phenotypes consisting of elevated ATP generation accompanied by increases in the levels of multiple intermediates of the tricarboxylic acid cycle, but lower α-ketoglutarate (α-KG) in MPN patients. Inhibition of PI3K/AKT/mTOR signaling significantly reduced metabolic responses and hyperreactivity in MPN patient platelets, while α-KG supplementation markedly reduced oxygen consumption and ATP generation. Ex vivo incubation of platelets from both MPN patients and Jak2 V617F–knockin mice with α-KG supplementation significantly reduced platelet activation responses. Oral α-KG supplementation of Jak2 V617F mice decreased splenomegaly and reduced hematocrit, monocyte, and platelet counts. Finally, α-KG treatment significantly decreased proinflammatory cytokine secretion from MPN CD14+ monocytes. Our results reveal a previously unrecognized metabolic disorder in conjunction with aberrant PI3K/AKT/mTOR signaling that contributes to platelet hyperreactivity in MPN patients.

Authors

Fan He, Angelo B.A. Laranjeira, Tim Kong, Shuyang Lin, Katrina J. Ashworth, Alice Liu, Nina M. Lasky, Daniel A.C. Fisher, Maggie J. Cox, Mary C. Fulbright, Lilian Antunes-Heck, LaYow Yu, Molly Brakhane, Bei Gao, Stephen M. Sykes, Angelo D’Alessandro, Jorge Di Paola, Stephen T. Oh

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Figure 8

α-KG inhibited monocyte activation and hyperinflammation.

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α-KG inhibited monocyte activation and hyperinflammation. (A) Plasma cyt...
(A) Plasma cytokine levels in HIs and MPN patients. Plasma from HIs (n = 9) and MPN patients (n = 28) was collected for the determination of 30 biomarkers using a V-PLEX Human Cytokine 30-Plex Kit from Meso Scale Discovery. Data are mean ± SD and were assessed by 2-tailed Mann-Whitney U test. MFpET, myelofibrosis post essential thrombocythemia. (B) Monocyte cytokine secretion changes with octyl-α-KG or oligomycin (Omy) treatment. Sorted CD14+ monocytes (0.5 × 106) from MPN patients (n = 5) were incubated with octyl-α-KG or Omy for 8 hours and the supernatants were collected for cytokine determination by multiplex Luminex assay. Data are mean ± SD and were assessed by 2-tailed Mann-Whitney U test. (C) The PCA score plot of RNA-seq of sorted CD14+ monocytes after the incubation with octyl-α-KG or DMSO control. (D) Bar plot of GSEA results showing top 5 hallmark pathways enriched in DMSO- versus α-KG–treated monocytes. (E) Heatmap showing changes in cytokine secretion by CD11b+ myeloid cells from Jak2 V617F mice. Data were normalized to control group as fold changes. Enriched CD11b+ myeloid cells were incubated with LPS (0.1 mg/mL) in the presence or absence of α-KG for 6 hours. Supernatants were collected for cytokine determination by multiplex Luminex assay. (F) Dot plots of altered intracellular pathways of monocytes in peripheral blood of MPN patients by mass cytometry. Whole blood from MPN patients (n = 6) were incubated with octyl-α-KG or DMSO for 1 hour followed by stimulation with TNF-α. Data are mean ± SD and were assessed by 2-tailed, paired Student’s t test. (G) Proposed model showing the roles of PI3K/AKT/mTOR signaling and metabolic changes in platelets from MPN patients. A positive feedback loop involving PI3K/AKT/mTOR signaling and metabolic changes promotes platelet hyperreactivities and megakaryopoiesis in MPN. The supplementation of α-KG, which disrupts the feedback loop, shows therapeutic effects against platelet hyperreactivity, megakaryopoiesis, and chronic inflammation in MPN.