EPO Receptor Gain-of-Function Causes Hereditary Polycythemia, Alters CD34+ Cell Differentiation and Increases Circulating Endothelial Precursors

Perrotta, Silverio; Cucciolla, Valeria; Ferraro, Marcella; Ronzoni, Luisa; Tramontano, Annunziata; Scudieri, Anna Chiara; Borriello, Adriana; Roberti, Domenico; Nobili, Brúno; Cappellini, Maria Domenica; Oliva, Adriana; Amendola, Giovanni; Migliaccio, Anna Rita; Mancuso, Patrizia; Martín‐Padura, Inés; Yoon, Donghoon; Prchal, Josef T.; Ragione, Fulvio Della

doi:10.1371/journal.pone.0012015

Cited by 25 publications

(40 citation statements)

References 41 publications

(50 reference statements)

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“…ingly, these mutations have been shown to lead to the accumulation of truncated EPOR at the plasma membrane, which is thought to underlie increased downstream signaling (24,25). Perhaps most notably, many of the disease-associated truncations eliminate Pro 419 (and Pro 426 ), which is predicted to cause a failure in the negative regulation of EPOR via VHL and a consequential prolongation of cell surface EPOR expression.…”

Section: Discussionmentioning

confidence: 99%

Oxygen-dependent Regulation of Erythropoietin Receptor Turnover and Signaling

Heir

Srikumar

Bikopoulos

et al. 2016

Journal of Biological Chemistry

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von Hippel-Lindau (VHL) disease is a rare familial cancer predisposition syndrome caused by a loss or mutation in a single gene, VHL, but it exhibits a wide phenotypic variability that can be categorized into distinct subtypes. The phenotypic variability has been largely argued to be attributable to the extent of deregulation of the ␣ subunit of hypoxia-inducible factor ␣, a well established target of VHL E3 ubiquitin ligase, ECV (Elongins/ Cul2/VHL). Here, we show that erythropoietin receptor (EPOR) is hydroxylated on proline 419 and 426 via prolyl hydroxylase 3. EPOR hydroxylation is required for binding to the ␤ domain of VHL and polyubiquitylation via ECV, leading to increased EPOR turnover. In addition, several type-specific VHL diseasecausing mutants, including those that have retained proper binding and regulation of hypoxia-inducible factor ␣, showed a severe defect in binding prolyl hydroxylated EPOR peptides. These results identify EPOR as the second bona fide hydroxylation-dependent substrate of VHL that potentially influences oxygen homeostasis and contributes to the complex genotypephenotype correlation in VHL disease.von Hippel-Lindau (VHL) 2 disease is a rare familial cancer predisposition syndrome characterized by the development of tumors in several organ systems, including the central nervous system and retinal hemangioblastoma (HAB), which represent the two cardinal features of VHL disease, as well as endolymphatic sac tumor of the inner ear, pancreatic cystadenoma, benign cystadenoma of epididymis, adnexal papillary cystadenoma of probable mesonephric origin, clear-cell renal cell carcinoma (RCC), paragangliomas, and pheochromocytoma (PHEO) (1). Some VHL patients also develop polycythemia with or without the other stigmata of VHL disease. VHL disease is subcategorized into type 1, characterized by HAB and RCC without PHEO; type 2A, characterized by PHEO and HAB without RCC; type 2B, characterized by PHEO and HAB with RCC; type 2C, characterized by exclusive development of PHEO; and type 3, which is characterized by the development of polycythemia without an increased cancer predisposition (2). Notably, despite the phenotypic heterogeneity, VHL disease is caused by a mutation or loss of a single gene, VHL.VHL is the substrate recognizing subunit of a multiprotein E3 ubiquitin ligase ECV (Elongins BC/Cul2/VHL) that targets the ␣ subunit of hypoxia-inducible factor (HIF␣) for ubiquitylation under normal oxygen tension (3). HIF␣ is hydroxylated on conserved prolines within the oxygen-dependent degradation domain (ODD) by a family of prolyl hydroxylase enzymes (PHD1-3) in the presence of 2-oxoglutarate, iron, and oxygen (4 -7). VHL binds exclusively to prolyl hydroxylated HIF␣. Thus, under hypoxia, the unmodified HIF␣ escapes the recognition by VHL and heterodimerizes with the constitutively expressed HIF␤ to form an active transcription factor, which transactivates numerous hypoxia-inducible genes involved in various adaptive processes to hypoxia such as anaerobic metabolism, angiogenesis, and er...

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Section: Discussionmentioning

confidence: 99%

Oxygen-dependent Regulation of Erythropoietin Receptor Turnover and Signaling

Heir

Srikumar

Bikopoulos

et al. 2016

Journal of Biological Chemistry

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“…8 Quantitative polymerase chain reaction analysis cDNA was prepared from patients' mRNA from erythroid precursors with an I-Script kit (Bio-Rad). Real-time polymerase chain reaction (PCR) was performed for adrenomedullin (ADM), N-myc downstream regulated gene 1 (NDRG1), vascular endothelial growth factor (VEGF) and transferrin receptor (TFR) genes, in accordance with the manufacturers' instructions.…”

Section: Erythroid Precursor Culturesmentioning

confidence: 99%

“…Erythroid precursors at specific stages of differentiation express high levels of EPO receptor (EPOR) whose engagement results in increased red cell production. [6][7][8] Induction of EPO gene transcription by hypoxia has led to the discovery of the transcription factor hypoxia-inducible factor (HIF) and the molecular pathways that allow the modulation of HIF levels by oxygen levels. 9 HIF consists of two subunits, HIF-α and HIF-β, whose nuclear heterodimerization results in the active form.…”

Section: Introductionmentioning

confidence: 99%

Congenital erythrocytosis associated with gain-of-function HIF2A gene mutations and erythropoietin levels in the normal range

et al. 2013

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Hypoxia-inducible factor 2α (HIF-2α) plays a pivotal role in the balancing of oxygen requirements throughout the body. The protein is a transcription factor that modulates the expression of a wide array of genes and, in turn, controls several key processes including energy metabolism, erythropoiesis and angiogenesis. We describe here the identification of two cases of familial erythrocytosis associated with heterozygous HIF2A missense mutations, namely Ile533Val and Gly537Arg. Ile533Val is a novel mutation and represents the genetic HIF2A change nearest to Pro-531, the primary hydroxyl acceptor residue, so far identified. The Gly537Arg missense mutation has already been described in familial erythrocytosis. However, our patient is the only described case of a de novo HIF2A mutation associated with the development of congenital polycythemia. Functional in vivo studies, based on exogenous expression of hybrid HIF-2α transcription factors, indicated that these genetic alterations lead to the stabilization of HIF-2α protein. All the identified polycythemic subjects with HIF2A mutations show serum erythropoietin in the normal range, independently of the hematocrit values and phlebotomy frequency. The erythroid precursors obtained from the peripheral blood of patients showed an altered phenotype, including an increased rate of growth and a modified expression of some HIF-2α target genes. These results suggest the novel proposal that polycythemia observed in subjects with HIF2A mutations might also be due to primary changes in hematopoietic cells and not only secondary to increased erythropoietin levels. Congenital erythrocytosis associated with gain-of-function HIF2A gene mutations and erythropoietin levels in the normal range

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“…Cytokine receptors that have no intrinsic kinase activity are also found mutated and overexpressed in hematopoietic disease and are believed to contribute to disease formation. For example, erythropoietin receptor (EpoR) mutations play a role in familial erythrocytosis/polycythemia [11–15] and granulocyte colony-stimulating factor (G-CSF) receptor (G-CSFR) mutations are found in AML as well as chronic neutrophilia [16–18]. Mutations of the thrombopoietin receptor, TpoR (Mpl), are found in myeloproliferative neoplasms (MPNs), including essential thrombocythemia and myelofibrosis, and mutated Mpl can signal to induce similar disease phenotypes in animal models [19, 20].…”

Section: Introductionmentioning

confidence: 99%

Mutations in the transmembrane and juxtamembrane domains enhance IL27R transforming activity

Lambert

Pradhan

Roll

et al. 2011

Biochemical Journal

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Synopsis Cytokines and their receptors regulate hematopoiesis by controlling cellular growth, survival, and differentiation. Thus, it is not surprising that mutations of cytokine receptors contribute to the formation of hematopoietic disorders including cancer. We recently identified transforming properties of the ligand-binding component of the receptor for interleukin-27 (IL27R). While wildtype IL27R exhibits transforming properties in hematopoietic cells, in this study we set out to determine if the transforming activity of IL27R could be enhanced by mutation. We identified three mutations of IL27R that enhance its transforming activity. One of these mutations is a phenylalanine to cysteine mutation at residue 523 (F523C) in the transmembrane domain of the receptor. The two other mutations identified involve deletions of amino acids in the cytoplasmic juxtamembrane region of the receptor. Expression of each of these mutant IL27R proteins led to rapid cytokine independent transformation in hematopoietic cells. Moreover, the rate of transformation induced by these mutants was significantly greater than that induced by wildtype IL27R. Expression of these IL27R mutants also induced enhanced activation of JAK/STAT signaling compared to wildtype. An activating deletion mutation of IL27R enhanced homodimerization of the receptor, by a mechanism that may involve disulfide bonding. These transforming IL27R mutants displayed equal or greater transforming activity than bona fide hematopoietic oncogenes such as BCR-ABL and JAK2-V617F. Since IL27R is expressed on hematopoietic stem cells, lymphoid cells, and myeloid cells, including AML blast cells, mutation of this receptor has the potential to contribute to a variety of hematopoietic neoplasms.

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EPO Receptor Gain-of-Function Causes Hereditary Polycythemia, Alters CD34+ Cell Differentiation and Increases Circulating Endothelial Precursors

Cited by 25 publications

References 41 publications

Oxygen-dependent Regulation of Erythropoietin Receptor Turnover and Signaling

Oxygen-dependent Regulation of Erythropoietin Receptor Turnover and Signaling

Congenital erythrocytosis associated with gain-of-function HIF2A gene mutations and erythropoietin levels in the normal range

Mutations in the transmembrane and juxtamembrane domains enhance IL27R transforming activity

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