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 7

α-KG exerts therapeutic effects on MPN and inhibits megakaryopoiesis.

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α-KG exerts therapeutic effects on MPN and inhibits megakaryopoiesis. (A...
(A) Schematic of the Jak2 V617F–knockin mice. cKit+ cells from Jak2 V617F CD45.2 C57BL/6J mice were isolated and transplanted into irradiated CD45.1 C57BL/6J mice. Two weeks after transplantation, mice were randomly grouped and supplemented with regular water (control, n = 10) or 1% α-KG in drinking water (n = 10) daily for 6 weeks. (B) Spleen weight of transplanted mice normalized to body weight measured at the end of treatments. Data are mean ± SD and were assessed by 2-tailed Student’s t test. (C) WBC, platelet (PLT) count, RBC, hematocrit (HCT), monocyte (MO) count, and ratio of Jak2 V617F–transplanted mice treated with regular water or α-KG across multiple time points. Data are mean ± SD and were assessed by 2-way ANOVA with Dunnett’s multiple-comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (D) Representative images of H&E staining of femur bones from mice treated with regular water or α-KG. (E) Representative images of immunofluorescent staining of expanded CD34+ cells and differentiated megakaryocytes from the same individual. Total original magnification, ×200 (D) and ×600 (E). (F) Flow cytometry of CD41 and CD61 surface expression on in vitro megakaryocytes differentiated with α-KG. Sorted CD34+ hematopoietic stem and progenitor cells were cultured for megakaryocyte differentiation with 250 μM octyl-α-KG or DMSO control. CD41+CD61+ double-positive cells were determined by flow cytometry. Data are mean ± SD and were assessed by 2-tailed, paired Student’s t test. (G) Percentage of LSK cells from Jak2 V617F–transplanted mice treated with regular water or α-KG at the end of treatments. Data are mean ± SD and were assessed by 2-tailed Student’s t test. (H) CFU assays of mouse cKit+ cells with DMSO control or α-KG. Colony numbers were counted after 14 days. Data are mean ± SD and were assessed by 2-tailed Student’s t test.