Multiomic profiling reveals metabolic alterations mediating aberrant platelet activity and inflammation in myeloproliferative neoplasms
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
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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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 5

Platelets from MPN patients displayed bioenergetic alterations, which can be reverted by α-KG supplementation.

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Platelets from MPN patients displayed bioenergetic alterations, which ca...
(A) Representative OCR and ECAR profiles of platelets from 2 HIs, 1 ET patient, and 1 MF patient (a, oligomycin A; b, FCCP; c, rotenone/antimycin A). (B) Quantification of basal OCR, ATP production, maximal OCR, and spare capacity profiles of washed platelets in HIs (n = 8) and MPN patients (n = 18: ET = 9, PV = 4, MF = 5). Data are mean ± SD and were assessed by Kruskal-Wallis test with Dunn’s multiple-comparison test. P values are marked if less than 0.05. (C) Representative OCR profiles of platelets from 1 HI and 1 ET patient showing the energy demand boost after TRAP6 stimulation and quantification of post-TRAP6 stimulation OCR profiles (a, oligomycin A; b, FCCP; c, rotenone/antimycin A). Data are mean ± SD and were assessed by Kruskal-Wallis test with Dunn’s multiple-comparison test. P values are marked if less than 0.05. (D) Correlation analysis of bioenergetic parameters and platelet functional parameters. (E) Representative OCR and ECAR profiles of platelets from 1 HI and 1 ET patient with the preincubation of 500 μM octyl-α-KG or DMSO for 1 hour (a, oligomycin A; b, FCCP; c, rotenone/antimycin A). (F) Platelet OCR/ECAR ratio from 1 HI and 1 ET patient with the preincubation of 500 μM octyl-α-KG or DMSO control. (G) Quantification of individual components of the platelet OCR profile in MPN (n = 5). Data were normalized to DMSO group set as 1, are presented as mean ± SD, and were assessed by 2-tailed Mann-Whitney U test.