Enhanced calreticulin expression in red cells of polycythemia vera patients harboring the <i>JAK2</i><sup>V617F</sup> mutation

Brusson, Mégane; Cochet, Sylvie; Guilhot, François; Mayeux, Patrick; Peyrard, Thierry; Chomienne, Christine; Kim, Caroline Le Van; Cassinat, Bruno; Kiladjian, Jean‐Jacques; Nemer, Wassim El

doi:10.3324/haematol.2016.161604

Cited by 12 publications

(22 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Three hundred and seventy-five proteins were confidently identified, with a number of peptides that allowed 358 distinct protein quantification ( Online Supplementary File 2 ). Among the deregulated proteins initially reported, 19 HC decreased the expression of several over-expressed proteins and increased the expression of those down-regulated ( Table 1 ). Nevertheless, the treatment did not seem to restore a normal expression pattern of these proteins for all 3 patients when compared to control ( Table 1 and Online Supplementary Table S1 ).…”

Section: Resultsmentioning

confidence: 95%

“…Our recent work showed abnormal expression of several proteins in RBCs of PV patients, 19 revealing qualitative alterations in these cells in addition to their increased number in the circulation. The aim of the current study was to address the effects of HC treatment on the expression of erythroid membrane proteins of PV patients using a global proteomic approach.…”

Section: Discussionmentioning

confidence: 91%

“…In our recent work, we showed abnormal expression of several proteins at the membrane of PV RBCs, with a high number of proteins from the endoplasmic reticulum, including calreticulin (Calr). 19 To verify if HC treatment restores normal expression of these proteins, we performed a proteomic analysis with RBC ghosts from 3 patients before and during their treatment with HC (pre-post patients). Three hundred and seventy-five proteins were confidently identified, with a number of peptides that allowed 358 distinct protein quantification ( Online Supplementary File 2 ).…”

Section: Resultsmentioning

confidence: 99%

“…In the group of membrane proteins abnormally expressed in PV RBCs, 19 Lu/BCAM was the only protein whose overexpression was further exacerbated by HC in all 3 patients, with a fold increase comprised between 6.6 and 11.8 ( Table 1 ). Lu/BCAM is a low abundance surface protein that is expressed on a subpopulation of circulating RBCs.…”

Section: Resultsmentioning

confidence: 99%

“… 17 , 18 More recently, using a proteomic approach, we also found that membranes of PV RBCs had abnormal expression of several proteins including the adhesion protein Lu/BCAM (Lutheran/Basal Cell Adhesion Molecule) and proteins from the endoplasmic reticulum, such as calreticulin (Calr) and calnexin. 19 These findings indicate that JAK2V617F not only impacts cell proliferation, but also induces changes in the repertoire of erythroid proteins expressed in circulating RBCs, which might contribute to the circulatory complications described in PV.…”

Section: Introductionmentioning

confidence: 81%

See 4 more Smart Citations

Impact of hydroxycarbamide and interferon-α on red cell adhesion and membrane protein expression in polycythemia vera

et al. 2018

Self Cite

View full text Add to dashboard Cite

Polycythemia vera is a chronic myeloproliferative neoplasm characterized by the JAK2V617F mutation, elevated blood cell counts and a high risk of thrombosis. Although the red cell lineage is primarily affected by JAK2V617F, the impact of mutated JAK2 on circulating red blood cells is poorly documented. Recently, we showed that in polycythemia vera, erythrocytes had abnormal expression of several proteins including Lu/BCAM adhesion molecule and proteins from the endoplasmic reticulum, mainly calreticulin and calnexin. Here we investigated the effects of hydroxycarbamide and interferon-α treatments on the expression of erythroid membrane proteins in a cohort of 53 patients. Surprisingly, while both drugs tended to normalize calreticulin expression, proteomics analysis showed that hydroxycarbamide deregulated the expression of 53 proteins in red cell ghosts, with overexpression and downregulation of 37 and 16 proteins, respectively. Within over-expressed proteins, hydroxycarbamide was found to enhance the expression of adhesion molecules such as Lu/BCAM and CD147, while interferon-α did not. In addition, we found that hydroxycarbamide increased Lu/BCAM phosphorylation and exacerbated red cell adhesion to its ligand laminin. Our study reveals unexpected adverse effects of hydroxycarbamide on red cell physiology in polycythemia vera and provides new insights into the effects of this molecule on gene regulation and protein recycling or maturation during erythroid differentiation. Furthermore, our study shows deregulation of Lu/BCAM and CD147 that are two ubiquitously expressed proteins linked to progression of solid tumors, paving the way for future studies to address the role of hydroxycarbamide in tissues other than blood cells in myeloproliferative neoplasms.

show abstract

Section: Resultsmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 91%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 81%

See 3 more Smart Citations

Impact of hydroxycarbamide and interferon-α on red cell adhesion and membrane protein expression in polycythemia vera

et al. 2018

Self Cite

View full text Add to dashboard Cite

show abstract

Blocking the CD47-SIRPα interaction reverses the disease phenotype in a polycythemia vera mouse model

et al. 2023

View full text Add to dashboard Cite

Polycythemia vera (PV) is a hematopoietic stem cell neoplasm driven by somatic mutations in JAK2, leading to increased red blood cell (RBC) production uncoupled from mechanisms that regulate physiological erythropoiesis. At steady-state, bone marrow macrophages promote erythroid maturation, whereas splenic macrophages phagocytose aged or damaged RBCs. The binding of the anti-phagocytic (“don’t eat me”) CD47 ligand expressed on RBCs to the SIRPα receptor on macrophages inhibits phagocytic activity protecting RBCs from phagocytosis. In this study, we explore the role of the CD47-SIRPα interaction on the PV RBC life cycle. Our results show that blocking CD47-SIRPα in a PV mouse model due to either anti-CD47 treatment or loss of the inhibitory SIRPα-signal corrects the polycythemia phenotype. Anti-CD47 treatment marginally impacted PV RBC production while not influencing erythroid maturation. However, upon anti-CD47 treatment, high-parametric single-cell cytometry identified an increase of MerTK+ splenic monocyte-derived effector cells, which differentiate from Ly6Chi monocytes during inflammatory conditions, acquire an inflammatory phagocytic state. Furthermore, in vitro, functional assays showed that splenic JAK2 mutant macrophages were more “pro-phagocytic,” suggesting that PV RBCs exploit the CD47-SIRPα interaction to escape innate immune attacks by clonal JAK2 mutant macrophages.

show abstract

Macrophage Inflammation, Erythrophagocytosis, and Accelerated Atherosclerosis in Jak2 ^V617F Mice

Wang

Liu

Fidler

et al. 2018

Circulation Research

198

133

View full text Add to dashboard Cite

Rationale: The mechanisms driving athero-thrombotic risk in individuals with JAK2V617F (Jak2VF) positive clonal hematopoiesis (CH) or myeloproliferative neoplasms (MPN) are poorly understood. Objective: The goal of this study was to assess atherosclerosis and underlying mechanisms in hypercholesterolemic mice with hematopoietic Jak2VF expression. Methods and Results: Irradiated low-density lipoprotein receptor knockout (Ldlr−/−) mice were transplanted with bone marrow from WT or Jak2VF mice and fed a high fat high cholesterol Western diet (WD). Hematopoietic functions and atherosclerosis were characterized. After 7 weeks of WD Jak2VF mice showed increased atherosclerosis. Early atherosclerotic lesions showed increased neutrophil adhesion and content, correlating with lesion size. After 12 weeks of WD Jak2VF lesions showed increased complexity, with larger necrotic cores, defective efferocytosis, prominent iron deposition and co-staining of erythrocytes and macrophages suggesting erythrophagocytosis. Jak2VF erythrocytes were more susceptible to phagocytosis by WT macrophages and showed decreased surface expression of CD47, a “don’t eat me” signal. Human JAK2VF erythrocytes were also more susceptible to erythrophagocytosis. Jak2VF macrophages displayed increased expression and production of pro-inflammatory cytokines and chemokines, prominent inflammasome activation, increased p38 MAP kinase signaling and reduced levels of MerTK, a key molecule mediating efferocytosis. Increased erythrophagocytosis also suppressed efferocytosis. Conclusions: Hematopoietic Jak2VF expression promotes early lesion formation and increased complexity in advanced atherosclerosis. In addition to increasing hematopoiesis and neutrophil infiltration in early lesions, Jak2VF caused cellular defects in erythrocytes and macrophages, leading to increased erythrophagocytosis but defective efferocytosis. These changes promote accumulation of iron in plaques and increased necrotic core formation which, together with exacerbated pro-inflammatory responses, likely contribute to plaque instability.

show abstract

Enhanced calreticulin expression in red cells of polycythemia vera patients harboring the JAK2^V617F mutation

Cited by 12 publications

References 16 publications

Impact of hydroxycarbamide and interferon-α on red cell adhesion and membrane protein expression in polycythemia vera

Impact of hydroxycarbamide and interferon-α on red cell adhesion and membrane protein expression in polycythemia vera

Blocking the CD47-SIRPα interaction reverses the disease phenotype in a polycythemia vera mouse model

Macrophage Inflammation, Erythrophagocytosis, and Accelerated Atherosclerosis in Jak2 ^V617F Mice

Contact Info

Product

Resources

About

Enhanced calreticulin expression in red cells of polycythemia vera patients harboring the JAK2V617F mutation

Cited by 12 publications

References 16 publications

Impact of hydroxycarbamide and interferon-α on red cell adhesion and membrane protein expression in polycythemia vera

Impact of hydroxycarbamide and interferon-α on red cell adhesion and membrane protein expression in polycythemia vera

Blocking the CD47-SIRPα interaction reverses the disease phenotype in a polycythemia vera mouse model

Macrophage Inflammation, Erythrophagocytosis, and Accelerated Atherosclerosis in Jak2 V617F Mice

Contact Info

Product

Resources

About

Enhanced calreticulin expression in red cells of polycythemia vera patients harboring the JAK2^V617F mutation

Macrophage Inflammation, Erythrophagocytosis, and Accelerated Atherosclerosis in Jak2 ^V617F Mice