RESUMO
Impaired protein homeostasis, though well established in age-related disorders, has been recently linked with the pathogenesis of myeloproliferative neoplasms (MPNs). However, little is known about MPN-specific modulators of proteostasis, thus impeding our ability for increased mechanistic understanding and discovery of additional therapeutic targets. Loss of proteostasis, in itself, is traced to dysregulated mechanisms in protein folding and intracellular calcium signaling at the endoplasmic reticulum (ER). Here, using ex vivo and in vitro systems (including CD34+ cultures from patient bone marrow and healthy cord/peripheral blood specimens), we extend our prior data from platelet RNA sequencing in patients with MPN and discover select proteostasis-associated markers at RNA and/or protein levels in each of platelet, parent megakaryocyte, and whole blood specimens. Importantly, we identify a novel role in MPNs for enkurin (ENKUR), a calcium mediator protein originally implicated only in spermatogenesis. Our data reveal consistent ENKUR downregulation at both RNA and protein levels across specimens from patients with MPN and experimental models (including upon treatment with thapsigargin, an agent that causes protein misfolding in the ER by selective loss of calcium), with a concomitant upregulation of a cell cycle marker, CDC20. Silencing of ENKUR using short hairpin RNA in CD34+-derived megakaryocytes further confirms this association with CDC20 at both RNA and protein levels and indicates a likely role for the PI3K/Akt pathway. Together, our work sheds light on enkurin as a novel marker of MPN pathogenesis and indicates further mechanistic investigation into a role for dysregulated calcium homeostasis and ER and protein folding stress in MPN transformation.
Assuntos
Transtornos Mieloproliferativos , Neoplasias , Masculino , Humanos , Megacariócitos/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Cálcio/metabolismo , Plaquetas/metabolismo , Transtornos Mieloproliferativos/genética , Antígenos CD34/metabolismo , Neoplasias/metabolismo , Proteínas de Ligação a Calmodulina/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismoRESUMO
Impaired protein homeostasis, though well established in age-related disorders, has been linked in recent research with the pathogenesis of myeloproliferative neoplasms (MPNs). As yet, however, little is known about MPN-specific modulators of proteostasis, thus impeding our ability for increased mechanistic understanding and discovery of additional therapeutic targets. Loss of proteostasis, in itself, is traced to dysregulated mechanisms in protein folding and intracellular calcium signaling at the endoplasmic reticulum (ER). Here, using ex vivo and in vitro systems (including CD34 + cultures from patient bone marrow, and healthy cord/peripheral blood specimens), we extend our prior data from MPN patient platelet RNA sequencing, and discover select proteostasis-associated markers at RNA and/or protein levels in each of platelets, parent megakaryocytes, and whole blood specimens. Importantly, we identify a novel role in MPNs for enkurin ( ENKUR ), a calcium mediator protein, implicated originally only in spermatogenesis. Our data reveal consistent ENKUR downregulation at both RNA and protein levels across MPN patient specimens and experimental models, with a concomitant upregulation of a cell cycle marker, CDC20 . Silencing of ENKUR by shRNA in CD34 + derived megakaryocytes further confirm this association with CDC20 at both RNA and protein levels; and indicate a likely role for the PI3K/Akt pathway. The inverse association of ENKUR and CDC20 expression was further confirmed upon treatment with thapsigargin (an agent that causes protein misfolding in the ER by selective loss of calcium) in both megakaryocyte and platelet fractions at RNA and protein levels. Together, our work sheds light on enkurin as a novel marker of MPN pathogenesis beyond the genetic alterations; and indicates further mechanistic investigation into a role for dysregulated calcium homeostasis, and ER and protein folding stress in MPN transformation. Key Points: Enkurin, a calcium adaptor protein, is identified as a novel marker of pathogenesis in MPNs.MPN megakaryocyte and platelet expression of enkurin at RNA and protein levels is inversely associated with a cell differentiation cycle gene, CDC20.Likely role for dysregulated calcium homeostasis, and ER and protein folding stress in MPN transformation.
RESUMO
Predicting disease progression remains a particularly challenging endeavor in chronic degenerative disorders and cancer, thus limiting early detection, risk stratification, and preventive interventions. Here, profiling the three chronic subtypes of myeloproliferative neoplasms (MPNs), we identify the blood platelet transcriptome as a proxy strategy for highly sensitive progression biomarkers that also enables prediction of advanced disease via machine-learning algorithms. The MPN platelet transcriptome reveals an incremental molecular reprogramming that is independent of patient driver mutation status or therapy. Subtype-specific markers offer mechanistic and therapeutic insights, and highlight impaired proteostasis and a persistent integrated stress response. Using a LASSO model with validation in two independent cohorts, we identify the advanced subtype MF at high accuracy and offer a robust progression signature toward clinical translation. Our platelet transcriptome snapshot of chronic MPNs demonstrates a proof-of-principle for disease risk stratification and progression beyond genetic data alone, with potential utility in other progressive disorders.
Assuntos
Biomarcadores Tumorais/genética , Plaquetas/metabolismo , Policitemia Vera/genética , Mielofibrose Primária/genética , Proteostase/genética , Trombocitemia Essencial/genética , Transcriptoma , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/metabolismo , Plaquetas/patologia , Reprogramação Celular , Criança , Pré-Escolar , Estudos de Coortes , Diagnóstico Diferencial , Progressão da Doença , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Policitemia Vera/diagnóstico , Policitemia Vera/metabolismo , Policitemia Vera/patologia , Mielofibrose Primária/diagnóstico , Mielofibrose Primária/metabolismo , Mielofibrose Primária/patologia , Medição de Risco , Trombocitemia Essencial/diagnóstico , Trombocitemia Essencial/metabolismo , Trombocitemia Essencial/patologiaRESUMO
BACKGROUND: Acute myeloid leukemia (AML) and advanced systemic mastocytosis (SM) are clonal diseases of the blood. Prognoses for patients with FMS-like tyrosine kinase 3 (FLT3) mutation-positive AML and those with advanced SM are poor. In the United States, midostaurin was approved in 2017 in combination with standard chemotherapy in patients with newly diagnosed FLT3 mutation-positive AML and as a single agent in patients with advanced SM. OBJECTIVES: This article aims to improve oncology nurses' knowledge about the benefits and risks of midostaurin therapy and to provide guidance on the identification and management of eligible patients. METHODS: The clinical data that supported the U.S. Food and Drug Administration's approval of midostaurin are reviewed, and supporting safety and management considerations are provided based on the authors' experiences as nurses and advanced practice providers caring for patients who received midostaurin during these key clinical trials. FINDINGS: Nausea and vomiting are the most frequent nonhematologic adverse events reported with midostaurin; therefore, administer midostaurin with antiemetics, and recommend taking it with food. Care should be taken when midostaurin is coadministered with strong CYP3A4 inhibitors.