Microarray analysis reveals distinct immune signatures in BCR?ABL positive and negative myeloproliferative neoplasms

Background: BCR?ABL mutation on the Philadelphia chromosome is the key driver of chronic myeloid leukemia (CML) pathogenesis. However, there are certain cases of myeloproliferative neoplasms (MPN) wherein no inherent driver mutation is detected resulting in clinical phenotype. It is important to identify key genes and pathways in driving the disease. The aim of the study was to use a gene-based omics approach to molecularly characterize these mutation-positive and negative cases to further strengthen diagnostics and precision medicine. Methods: A microarray profiling was done on CD34 positive cells isolated from two BCR?ABL positive and five BCR?ABL negative samples. JAK2V617F mutation testing was also done to rule out the presence of any other mutation in the latter group. The fold change cut-off was taken as �5 with p?0.5 for significant genes. The gene network and pathway analysis were done using DAVID and STRING software. Results: The genes upregulated in BCR?ABL negative samples were shown to be involved in immune regulation, signal transduction and T- and B-cell signalling. The protein-protein interaction network of upregulated genes in these samples were enriched for various immunomodulatory genes such as HLADP, HLADQ , IL7R, CCR7, CD3 subtypes. These genes further formed a network with signal transduction genes such as LCK, FYN, RAG1, DOCK1, AKT3, SMAD3, LEF1. Conclusion: The results suggested a modulation of immune response genes and its subsequent effect on oncogenic signalling in BCR?ABL negative samples as compared to BCR?ABL positive samples. The protein network analysis was enriched for genes involved in Src, TGF-beta and PI3K-AKT pathway contributing to the proliferation of neoplastic clone.

Efficacy and safety of peginterferon-α2b for treatment of myeloproliterative neoplasms / 南方医科大学学报

OBJECTIVE@#To evaluate the clinical efficacy and adverse reactions of peginterferon-α2b for treatment of chronic myeloproliferative neoplasms (MPN).@*METHODS@#We retrospectively analyzed the data of 107 patients with MPN, including 95 with essential thrombocythemia (ET) and 12 with polycythemia vera (PV), who all received peginterferon-α2b treatment for at least 12 months. The clnical and follow-up data of the patients were analyzed to evaluate the efficacy and adverse reactions of the treatment.@*RESULTS@#After receiving peginterferon- α2b treatment, both ET and PV patients achieved high hematological remission rates, and the total remission rates did not differ significantly between the two groups (86% vs 78%, P>0.05). In the overall patients, the spleen index decreased by 13.5% (95%CI: 8.5%-18.5%) after the treatment. The patients with hematological remission showed a significantly greater reduction of the total symptom score than those without hematological remission (P < 0.01). The median percentage of JAK2V617F allele load of PV patients decreased from 67.23% (49.6%-84.86%) at baseline to 19.7% (0.57%-74.6%) after the treatment, and that of JAK2V617F-positive ET patients decreased from 48.97% (0.45%-74.24%) at baseline to 22.1% (0.33%-65.42%) after the treatment. Mild adverse reactions (grade 1-2) were observed in both ET and PV groups without significant differences between them. The overall incidence of thrombotic events during the treatment was 2.8% in these patients, and no serious adverse reactions were observed.@*CONCLUSION@#For patients with chronic myelodysplasia, peginterferon-α2b treatment can achieve a high peripheral blood cell remission rate and maintain a long-term stable state with good effect in relieving symptoms such as splenomegaly. Peginterferon- α2b treatment caused only mild adverse reactions, which can be tolerated by most of the patients.

Focal adhesion kinase inhibition decreases cell viability and induces apoptosis of JAK2 V617F positive cells

Abstract Focal Adhesion Kinase (FAK) protein participates in proliferation, migration, cell survival, and apoptosis process. It has been described as overexpressed in several neoplasms being a promising target for therapy. BCR-ABL negative chronic Myeloproliferative Neoplasms (MPN) are clonal disorders characterized by the excess of proliferation and apoptosis resistance. The identification of the acquired JAK2 V617F mutation in MPN patients allowed a better understanding of pathogenesis. However, there is still no pharmacological treatment that leads all patients to molecular remission, justifying new studies. The present study aimed to evaluate FAK involvement in the viability and apoptosis of HEL and SET-2 cells, both JAK2 V617F positive cell lines. The FAK inhibitor PF 562,271 was used. Cell viability was determined using MTT assay and apoptosis verified by cleaved PARP, cleaved Caspase 3 and Annexin-V/PI staining detection. FAK inhibition significantly reduced HEL and SET-2 cells viability and induced apoptosis. Considering the role of JAK/STAT pathway in MPN, further investigation of FAK participation in the MPN cells proliferation and apoptosis resistance, as well as possible crosstalk between JAK and FAK and downstream pathways may contribute to the knowledge of MPN pathophysiology, the discovery of new molecular targets, and JAK inhibitors resistance mechanisms.

Calreticulin Mutations in Myeloproliferative Neoplasms Patients Diagnosed in UKM Medical Centre

@#Introduction: Calreticulin (CALR) mutations are one of the molecular markers that has been incorporated for the diagnosis of myeloproliferative neoplasms (MPN) in the revised 2017 WHO Classification of Haematopoietic and Lymphoid Tumors. This study was performed to determine the prevalence of CALR mutations in patients with MPN diagnosed in UKMMC and to compare their demographics plus laboratory features with other MPN patients. Methods: A total of 59 MPN patients who tested negative for JAK2V617Fmutation were selected and 21 MPN patients positive for JAK2V617F were included as controls. Screening for CALR exon 9 was done by multiplex polymerase chain reaction (PCR) followed by Sanger sequencing. Results: A total of six JAK2 V617F negative MPN samples were found to be positive for CALR mutations. Out of these six, three patients with CALR mutations were of type I mutation, two were type II while one was a mutation in the stretch III region. None of the twenty one JAK2 V617F positive MPN samples were positive for CALR mutation. Clinical phenotypes for those positive for CALR were restricted to Essential Thrombocythemia (ET), Primary Myelofibrosis (PMF) and one case of atypical Chronic Myeloid Leukaemia (CML). Conclusion: CALR mutations constituted 10.16% from the MPN patients who were negative for JAK2V617F mutation with no significant differences in platelet counts, hemoglobin (Hb), hematocrit and white cell counts as compared to MPN patients with JAK2 V617F mutations. Testing for CALR mutations among those who are negative for JAK2V617F within Malaysian population maybe worthwhile and require larger scale studies.

Myeloproliferative Neoplasm Symptom Assessment Form - Total Symptom Score (MPN-SAF TSS) questionnaire: translation, cultural adaptation and validation to Brazilian Portuguese

ABSTRACT Introduction: Constitutional symptoms and thrombohemorrhagic events are common in patients with myeloproliferative neoplasms (MPNs). Hence, the treatment's primary goal is to control symptoms and improve the quality of life (QoL). In order to assess response to therapy, symptom burden, and QoL among patients with MPN, the "Myeloproliferative Neoplasm Symptom Assessment Form - Total Symptom Score (MPN-SAF TSS)" questionnaire was developed in the USA in 2012. Herein, we translated and validated the MPN-SAF TSS questionnaire to Brazilian Portuguese. Methods: The ten-item questionnaire was translated from the English language and its psychometric properties (reliability, convergent and construct validities) were evaluated in 101 MPN patients. Results: There were 41 patients with essential thrombocythemia, 39 with myelofibrosis and 21 with polycythemia vera. The median age of all patients at diagnosis was 68 years and 59% were female. The Cronbach's alpha coefficient for the overall questionnaire was 0.78, ranging from 0.73 to 0.79, if each item was deleted. Validity analyses showed that the strongest item-item correlation were between early satiety and abdominal discomfort. Strong correlations were also found between physician and patient perceptions of itching (r = 0.81) and fatigue (r = 0.70). The Pearson coefficient correlation between the MPN-SAF TSS global score and the EORTC QLQ-C30 functional scales ranged from 0.51 to 0.64. The exploratory factor analysis showed that seven of the ten symptoms loaded into one single factor. Conclusion: The Brazilian Portuguese version of the MPN-SAF-TSS showed good psychometric properties and can be an available tool to assess symptom burden in this group of patients.

Progress of BCR-ABL fusion gene and JAK2 V617F mutation double-positive myeloproliferative neoplasms / 白血病·淋巴瘤

Myeloproliferative neoplasms (MPN) are a group of clonal disorders of hematopoietic stem cells, and JAK2 V617F gene mutation is the main basis for the diagnosis of MPN. Previous studies have shown that BCR-ABL fusion gene and JAK2 V617F gene mutation are mutually exclusive in MPN patients, but in recent years, patients with a double mutation of both genes are often reported. The article synthesizes the relevant domestic and foreign literature in recent years, and reviews the BCR-ABL fusion gene and JAK2 V617F mutation double-positive MPN.

IRF4 and IRF8 expression are associated with clinical phenotype and clinico-hematological response to hydroxyurea in essential thrombocythemia / 医学前沿

The morbidity and mortality of myeloproliferative neoplasms (MPNs) are primarily caused by arterial and venous complications, progression to myelofibrosis, and transformation to acute leukemia. However, identifying molecular-based biomarkers for risk stratification of patients with MPNs remains a challenge. We have previously shown that interferon regulatory factor-8 (IRF8) and IRF4 serve as tumor suppressors in myeloid cells. In this study, we evaluated the expression of IRF4 and IRF8 and the JAK2V617F mutant allele burden in patients with MPNs. Patients with decreased IRF4 expression were correlated with a more developed MPN phenotype in myelofibrosis (MF) and secondary AML (sAML) transformed from MPNs versus essential thrombocythemia (ET). Negative correlations between the JAK2V617F allele burden and the expression of IRF8 (P < 0.05) and IRF4 (P < 0.001) and between white blood cell (WBC) count and IRF4 expression (P < 0.05) were found in ET patients. IRF8 expression was negatively correlated with the JAK2V617F allele burden (P < 0.05) in polycythemia vera patients. Complete response (CR), partial response (PR), and no response (NR) were observed in 67.5%,10%, and 22.5% of ET patients treated with hydroxyurea (HU), respectively, in 12 months. At 3 months, patients in the CR group showed high IRF4 and IRF8 expression compared with patients in the PR and NR groups. In the 12-month therapy period, low IRF4 and IRF8 expression were independently associated with the unfavorable response to HU and high WBC count. Our data indicate that the expression of IRF4 and IRF8 was associated with the MPN phenotype, which may serve as biomarkers for the response to HU in ET.

Myelodysplastic syndrome/myeloproliferative neoplasm with ring sideroblasts and thrombocytosis: report of 2 cases and review of literature / 白血病·淋巴瘤

Objective:To deepen the understanding of myelodysplastic syndrome/myeloproliferative neoplasm with ring sideroblasts and thrombocytosis (MDS/MPN-RS-T), and to improve the levels of precise diagnosis and individualized treatment.Methods:The clinical data and next-generation sequencing molecular cloning results of two MDS/MPN-RS-T patients who were admitted to the First People's Hospital of Chuzhou in October 2017 and November 2019 were retrospectively analyzed, and the related literature was reviewed.Results:Case 1 was a 76-year-old female. The mutation loads from high to low were DNMT3A, JAK2 V617F and SF3B1. After administration of hydroxyurea, this patient acquired amelioration in anemia, and the platelet count improved. The clinical course was indolent. Case 2 was a 66-year-old male, who was initially diagnosed with essential thrombocythemia but failed to acquire response after hydroxyurea treatment. MDS/MP-RS-T was diagnosed after comprehensive examination. The mutation loads from high to low were SF3B1, ASXL1, JAK2 V617F and SRSF2. Pancytopenia occurred after disease progression, and the JAK2 V617F mutation finally turned negative. Administration of erythropoietin and lenalidomide failed to improve the condition, but low-dose decitabine treatment (10 mg/d, 3-5 d, once a month) improved the hematopoiesis.Conclusions:The co-mutation of JAK2 V617F and SF3B1 has a suggestive effect on the diagnosis of MDS/MPN-RS-T, and dynamic next-generation sequencing is helpful to elucidate the molecular nature of clinical heterogeneity of the disease. Low-dose decitabine has a certain curative effect on MDS/MPN-RS-T.

Manejo anestésico de pacientes con trombocitemia esencial / Anesthetic management of a patient with essential trombocythemia

La trombocitemia esencial forma parte del grupo de neoplasias mieloproliferativas. Se caracteriza por síntomas microvasculares y vasomotores, recuento plaquetario superior a 450 x 109/l, proliferación megacariocítica con morfología grande y madura, ausencia de proliferación eritroide y granulocítica, demostración de JAK2V617F u otro marcador clonal y ausencia de evidencia de trombocitosis reactiva. Se reporta el manejo anestésico en una paciente donde las principales consideraciones están relacionadas con la prevención de eventos hemorrágicos y trombóticos. La suspensión de la aspirina, el mantenimiento del tratamiento con hidroxiurea, la preparación con ácido tranexámico, el uso pre y posoperatorio de fraxiparina, hidratación adecuada, uso de medias elásticas en miembros inferiores, deambulación precoz, buena hemostasia quirúrgica y disponibilidad de concentrados de plaquetas son los elementos fundamentales en la conducción anestésica de esta paciente(AU)

Essential thrombocythemia is part of the group of myeloproliferative neoplasms. It is characterized by microvascular and vasomotor symptoms, platelet count over 450x109/L, megakaryocytic proliferation with large and mature morphology, absence of erythroid and granulocytic proliferation, demonstration of JAK2V617F or other clonal marker, and absence of evidence of reactive thrombocytosis. Anesthetic management is reported in a patient, whose case's main considerations are related to the prevention of hemorrhagic and thrombotic events. Aspirin suspension, maintenance of hydroxyurea treatment, preparation with tranexamic acid, pre- and post-operative use of fraxiparin, adequate hydration, use of elastic stockings in lower limbs, early ambulation, good surgical hemostasis, as well as availability of platelet concentrates are the fundamental elements in the anesthetic management of this patient(AU)

Exploring redox vulnerabilities in JAK2V617F-positive cellular models

ABSTRACT Background: In Philadelphia chromosome-negative myeloproliferative neoplasm (MPN) models, reactive oxygen species (ROS) are elevated and have been implicated in genomic instability, JAK2/STAT signaling amplification, and disease progression. Although the potential effects of ROS on the MPN phenotype, the effects of ruxolitinib treatment on ROS regulation have been poorly explored. Herein, we have reported the impact of ruxolitinib on redox signaling transcriptional network, and the effects of diphenyleneiodonium (DPI), a pan NOX inhibitor, in JAK2V617F-driven cellular models. Method: Redox signaling-related genes were investigated in SET2 cells upon ruxolitinib treatment by RNA-seq (GEO accession GSE69827). SET2 and HEL cells, which represent JAK2V617F-positive MPN cellular models with distinct sensitivity to apoptosis induced by ruxolitinib, were used. Cell viability was evaluated by MTT, apoptosis by annexin V/PI and flow cytometry, and cell signaling by quantitative PCR and Western blot. Main results: Ruxolitinib impacted on a network composed of redox signaling-related genes, and DUOX1 and DUOX2 were identified as potential modulators of ruxolitinib response. In SET2 and HEL cells, DPI reduced cell viability and, at low doses, it significantly potentiated ruxolitinib-induced apoptosis. In the molecular scenario, DPI inhibited STAT3, STAT5 and S6 ribosomal protein phosphorylation and induced PARP1 cleavage in JAK2V617F-positive cells. DPI combined with ruxolitinib increased PARP1 cleavage in SET2 cells and potentiated ruxolitinib-reduced STAT3, STAT5 and S6 ribosomal protein in HEL cells. Conclusion: Our study reveals a potential adaptation mechanism for resistance against ruxolitinib by transcriptionally reprogramming redox signaling in JAK2V617F cells and exposes redox vulnerabilities with therapeutic value in MPN cellular models.

Guidelines for therapy of patients with chronic myeloproliferative neoplasms during the novel coronavirus SARS-CoV2 pandemic

ABSTRACT The novel coronavirus has swept across the world in 2020 and ushered a new era. In the current scenario, it is not clear how patients with myeloproliferative neoplasms (including chronic myelogenous leukemia) should be managed, considering the risk of therapy, the need for social distancing and the risk of untimely therapy discontinuation of delay. This guideline aims to give providers a sense of direction in order to better take care of patients and prioritize care.

Comparison Study between Real Life and Published Data Outcome among Myelofibrosis Patients Who are Using Ruxolitinib at King Abdulaziz Medical City-Central Region

Objectives: The aim of this study is to evaluate the efficacy and safety of Ruxolitinib among Myelofibrosis patients in real life compared to published data. Methods: In this retrospective observational chart review study we reviewed the medical records for all patients diagnosed with Myelofibrosis in King Abdulaziz Medical City who has received Ruxolitinib from March 1, 2012-December 1, 2015. The efficacy and safety results of Ruxolitinib were compared with published data. Results: A total of 20 patients were included. The average age was 63.3 (SD=11.6) years, with 55% of females. Efficacy: At week 24, only 20% of the study participant achieved spleen size reduction equal to or more than 20% with total average of 8% reduction in the spleen size as compared to 31.60% in COMFORT-1 study (p ≤ 0.001), The highest symptoms reduction observed with fatigue and bone pain (45% and 40% of the affected patients respectively) followed by Abdominal distress (35%) whereas no statistically significant difference observed in early satiety and night sweat. Weight loss showed improvement in 15% of the patients. Safety: Fatigue was observed in 45% of the patients, diarrhea (5%), dyspnea (15%), dizziness (5%), nausea (5%), constipation (10%), vomiting (5%), pain in extremities (5%), arthralgia (5%), pyrexia (5%), and abdominal pain (35%). At week 24, Platelet count decreased by 26% and hemoglobin decreased by 5% from the baseline. In general, only three reported cases for temporal or permanent drug discontinuation. Conclusion: Ruxolitinib therapy in real life as compared to published trials was associated with significant improvement in Myelofibrosis related symptoms and splenomegaly with an acceptable safety profile

The genetic characteristics of BCR-ABL-negative myeloproliferative neoplasms / 中华内科杂志

Objective@#To explore the relationship between driver gene mutation (JAK2, MPL and CALR) and disease type in BCR-ABL negative myeloproliferative neoplasms (MPNs) including primary myeloid fibrosis (PMF), essential thrombocytosis (ET) and polycythemia vera (PV).@*Methods@#A total of 32 MPN related genes were detected by high-throughput sequencing in 156 MPN patients. The relationships between disease type and patients′ general performance, the characteristics of driver gene mutations, concomitant gene mutations were analyzed.@*Results@#In the population with JAK2 V617F positive mutation, the proportion of patients over 60 years old in PMF was higher than that with ET or PV. By high-throughput sequencing, 22 concomitant gene mutations were detected in 46 patients with JAK2, MPL or CALR mutations, including 4 (8.3%) in PV, 20 (29.4%) in ET, and 22 (55.0%) in PMF. DNMT3A mutation was detected only in patients with PV, while splicing factor related genes including SF3B1, SRSF2 and U2AF1 were only accompanied by PMF. According to the variation allele frequency (VAF) value of JAK2 V617F mutation, the VAF value associated with PV was the highest (68.15%), followed by PMF (37.7%) and ET (23%). However, there were significant differences in the incidence of JAK2 V617F homozygous among 3 different diseases. In patients with JAK2 mutation, the proportion of other gene mutations in PV and ET was significantly lower than that in PMF.@*Conclusions@#Under the condition of common driver gene mutations (JAK2, MPL and CALR), patients′ age, VAF value and homozygous state, concomitant gene mutations are closely related to different disease type. These correlations help to improve clinical understanding of disease characteristics and risk assessment.

Biomarcadores en las neoplasias mieloproliferativas clásicas BCR-ABL1 negativas / Biomarkers in Classical Myeloproliferative Neoplasms BCR-ABL1 negative

Introducción: Los biomarcadores son útiles en la definición del diagnóstico, pronóstico y seguimiento de múltiples enfermedades. La detección o medición de uno o más biomarcadores específicos representan alteraciones en vías genéticas o epigenéticas que controlan la proliferación, diferenciación o muerte celular. Las neoplasias mieloproliferativas constituyen un grupo fenotípicamente diverso de hemopatías malignas de origen clonal, caracterizadas por una sobreproducción simple o multilineal de los elementos eritroides, mieloides y megacariocíticos; así como de una marcada predisposición a la trombosis, sangramiento y transformación leucémica. Dentro de ellas se incluyen: la policitemia vera, la trombocitemia esencial y la mielofibrosis primaria, conocidas como neoplasias mieloproliferativas clásicas BCR-ABL1 (o cromosoma Philadelfia) negativas. Las mutaciones somáticas en genes como JAK2, MPL y CARL se comportan como mutaciones drivers iniciadoras, responsables del fenotipo mieloproliferativo. Métodos: Se revisaron artículos relacionados publicados en los últimos años, en algunas bases de datos de la Biblioteca Virtual de Salud. En esta revisión se exponen los mecanismos moleculares generales de esas mutaciones y su expresión clínica; se hace referencia a las neoplasias mieloproliferativas triple negativas y sus implicaciones clínicas y se indica el algoritmo diagnóstico propuesto por la Organización Mundial de la Salud que incluye los nuevos biomarcadores. Conclusiones: El estudio molecular proporciona información valiosa para el diagnóstico y seguimiento de las neoplasias mieloprolifrativas, pero no logra diferenciar entre cada una de ellas. Por esto, se requiere de la adecuada aplicación del método clínico para llegar a un diagnóstico certero con ayuda de otros exámenes complementarios(AU)

Introduction: Biomarkers are useful in the definition of diagnosis, prognosis and monitoring of multiple diseases. The detection or measurement of one or more specific biomarkers represents alterations in genetic or epigenetic pathways that control proliferation, differentiation or cell death. The myeloproliferative neoplasms constitute a phenotypically diverse group of malignant hemopathies of clonal origin, characterized by a simple or multilinear overproduction of the erythroid, myeloid and megakaryocytic elements; as well as a marked predisposition to thrombosis, bleeding and leukemic transformation. These include: polycythemia vera, essential thrombocythemia, and primary myelofibrosis, known as classical negative myeloproliferative neoplasms BCR-ABL1 (or Philadelphia chromosome). Somatic mutations in genes such as JAK2, MPL and CARL behave as initiating driver mutations responsible for the myeloproliferative phenotype. Methods: Articles published in the last years were reviewed in some databases of the Virtual Health Library (VHL). In this review we expose the general molecular mechanisms of these mutations and their clinical expression; reference is made to the triple negative myeloproliferative neoplasms and their clinical implications and the diagnostic algorithm proposed by the World Health Organization that includes the new biomarkers is indicated. Conclusions: The molecular study provides valuable information for the diagnosis and monitoring of myeloproliferative neoplasms, but fails to differentiate between each of them. Therefore, the appropriate application of the clinical method is required to arrive at an accurate diagnosis with the help of other complementary tests(AU)

Calidad de vida en pacientes con neoplasias mieloproliferativas crónicas Filadélfia negativas / Quality of life in patients with Philadelphia negative chronic myeloproliferative neoplasms

Resumen Introducción: la carga sintomática de pacientes con neoplasias mieloproliferativas crónicas Filadélfia negativas (NMC-PhN) afecta la calidad de vida (CV). Existen escalas para evaluar la magnitud de los síntomas, una de ellas, MPN-SAF-10. En nuestra región existe escasa información sobre CV de pacientes con NMC-PhN. Objetivos: estimar el puntaje de calidad de vida con la escala MPN-SAF-10 en pacientes con NMC-PhN atendidos en el Hospital de San José (Bogotá, Colombia) y explorar asociaciones entre el tiempo de tratamiento, carga de complicaciones y el efecto en la CV. Material y métodos: estudio de corte transversal analítico para evaluar CV basada en carga sintomática de pacientes con NMC-PhN del Hospital de San José (Bogotá, Colombia). Se realizó análisis descriptivo y estratificado de calidad de vida, tratamiento citorreductor y de diferentes complicaciones, así como pruebas de asociación de los puntajes de riesgo de cada enfermedad con sus respectivos puntajes de CV. Resultados: en 64 pacientes la escala MPN-SAF-10 documentó medianas de puntajes globales de CV de 3 (RIC 1-6), MPN-SAF-10 de 20 (RIC 8-32). Un 49% de los pacientes tuvo algún grado de alteración (30% moderada y 19% severa), sin diferencias entre las tres enfermedades. Los puntajes de CV no variaron entre las NMC-PhN. El tratamiento y duración del mismo no se correlacionaron con la escala de MPN-SAF-10 (hidroxiúrea r: - 0.27; ruxolitinib r: 0.12). Conclusiones: en pacientes con NMC-PhN, la evaluación de CV con la escala MPN-SAF-10 evidencia algún grado de afectación a pesar del tratamiento; ésta es útil para objetivar dicha afectación y debe implementarse en la práctica clínica. (Acta Med Colomb 2019; 44: 82-90).

Abstract Introduction: the symptomatic burden of patients with Philadelphia negative chronic myelopro liferative neoplasms (NMC-PhN) affects the quality of life (QL). There are scales to evaluate the magnitude of the symptoms; one of them, MPN-SAF-10. In our region there is scarce information on QL of patients with NMC-PhN. Objectives: To estimate the quality of life score with the MPN-SAF-10 scale in patients with NMC-PhN treated at Hospital de San José (Bogotá, Colombia) and to explore associations between treatment time, complication load and the effect on the QL. Material and methods: Analytical cross-sectional study to evaluate QL based on symptomatic load of patients with NMC-PhN from Hospital de San José (Bogotá, Colombia). A descriptive and stratified analysis of quality of life, cytoreductive treatment and different complications was carried out, as well as association tests of the risk scores of each disease with their respective QL scores. Results: in 64 patients the MPN-SAF-10 scale documented medians of global QL scores of 3 (RIC 1-6), MPN-SAF-10 of 20 (RIC 8-32). 49% of the patients had some degree of alteration (30% moderate and 19% severe), without differences between the three diseases. The QL scores did not vary between the NMC-PhN. The treatment and its duration did not correlate with the MPN-SAF-10 scale (Hydroxyurea r: - 0.27, Ruxolitinib r: 0.12). Conclusions: in patients with NMC-PhN, the evaluation of QL with the MPN-SAF-10 scale shows some degree of affectation despite the treatment; this is useful to objectify this affectation and should be implemented in clinical practice. (Acta Med Colomb 2019; 44: 82-90).

Calreticulina: aspectos moleculares y técnicas para su detección / Calreticulin: molecular aspects and techniques for its detection

Las neoplasias mieloproliferativas crónicas (NMPC) son enfermedades clonales caracterizadas por un aumento en el número de células maduras circulantes; estas incluyen: policitemia vera (PV), trombocitemia esencial (TE), mielofibrosis primaria (MFP), entre otras. Una de las principales características moleculares de estas tres entidades es la ausencia del gen de fusión BCR/ABL. La primera mutación relacionada directamente con estas neoplasias fue detectada en el gen JAK2; a partir de su descubrimiento, otras mutaciones en los genes del receptor de trombopoyetina (MPL) y calreticulina (CALR) han sido fuertemente relacionadas con la presentación de la enfermedad. La calreticulina es una proteína del retículo endoplásmico con diversas funciones a nivel celular como la homeostasis del calcio y la actividad de chaperona. Hasta la fecha se ha identificado un gran número de mutaciones en el gen CALR. La mayoría de ellas son inserciones y deleciones que generan cambios a nivel proteico con implicaciones importantes en el curso clínico y pronóstico de las neoplasias. Debido a su alta frecuencia y fuerte asociación con las NMPC, las mutaciones de CALR se incluyen como criterio mayor para el diagnóstico de estas entidades. Por este motivo, se han desarrollado varias técnicas encaminadas a la detección rápida, eficiente, sensible y especifica de esta mutación como: la secuenciación, el análisis de fragmentos y el análisis de fusión de alta resolución. El conocimiento e implementación de estas técnicas en los laboratorios clínicos constituye un avance importante para el diagnóstico y la evolución de los pacientes(AU)

Chronic myeloproliferative neoplasms (NMPC) are clonal diseases characterized by an increase in the number of mature circulating cells; these diseases include: polycythemia vera (PV), essential thrombocythemia (ET), primary myelofibrosis (MFP) among others. One of the main molecular characteristics of these three entities is the absence of the BCR/ABL fusion gene. The first mutation related to this group of neoplasms was detected in the JAK2 gene; since its discovery, other mutations in thrombopoietin receptor (MPL) and calreticulin (CALR) genes have been strongly related with the presentation of the disease. Calreticulin is an endoplásmic reticulum protein with different functions in the cell such as calcium homeostasis and the chaperone activity. To date, a large number of mutations have been identified in CALR gene most of them are insertions and deletions that generate changes in the protein that generate important implications in the clinical course and prognosis of neoplasms. Due to its high frequency and strong association with NMPC, CALR mutations are included as a major criteria for the diagnosis of these entities. For this reason, several techniques have been developed aimed at the rapid, efficient, sensitive and specific detection of this mutation as: sequencing, fragment analysis and high resolution fusion analysis. The knowledge and implementation of these techniques in clinical laboratories is an important advance for the diagnosis and in the evolution of patients(AU)

Correlation analysis on JAK2V617F mutation and clinical features in patients with myeloproliferative neoplasms and its clinical significance / 吉林大学学报(医学版)

Objective: To investigate the distribution of JAK2V617F mutation in the patients with myeloproliferative neoplasms (MPN), to explore the association between JAK2V617F mutation and the clinical features of MPN, and to clarify its clinical significance in the diagnosis and treatment of the MPN patients. Methods: A total of 170 patients with MPN were selected as the subjects and divided into true polycythemia (PV) group (n=68), primary thrombocytopenia (ET) group (n=88) and primary myelofibrosis (PMF) group (n=14). The JAK2V617F mutations in 170 patients with MPN were detected by allele-specific PCR (AS-PCR). The differences in gender, age, white blood cell count, hemoglobin, platelet count, prothrombin time (PT), activated partial thromboplastin time (APTT), D-dimer, whether complicated with splenomegaly and thromboembolism of the JAK2V617F mutant-type and wild-type patients in each group were analyzed. Results: The total mutation rate of JAK2V617F mutation in 170 patients with MPN was 68. 2% (116/170), and the mutation rates in PV, ET, and PMF groups were 72. 1% (49/68), 68. 1% (60/88), and 50. 0% (7/14), respectively. The JAK2V617F mutation rate in PV group was higher than those in the other two groups, but there were no statistical differences (P = 0. 281). Compared with wild type group, the age of onset, the white blood cell and platelet counts of the PV patients with JAK2V617F mutation were significantly increased (P

Significance of calreticulin gene mutations classification in BCR-ABL1 negative myeloproliferative neoplasms / 白血病·淋巴瘤

Objective@#To investigate calreticulin (CALR) gene mutations classification in BCR-ABL1 negative myeloproliterative neoplasms (MPN), and its relationship with clinical manifestations.@*Methods@#Genomic DNA polymerase chain reaction (PCR) amplification product Sanger sequencing method was used to detect the mutation of exon 9 of CALR gene in 236 patients with BCR-ABL1 negative MPN (excluding polycythemia vera and negative CALR mutations) in Ruijin Hospital of Shanghai Jiao Tong University School of Medicine from November 2015 to November 2018. The mutations were classified into 52 bp deletion (type 1) mutation, 5 bp insertion (type 2) mutation and other mutation types according to PCR sequencing analysis. The clinical characteristics of the carriers with two kinds of mutations in 198 patients with essential thrombocythemia (ET) and 38 primary myelofibrosis (PMF) were compared. For the types of mutations that could not be determined, they were classified according to the α-helix propensity score of the mutant protein peptide chain or the degree of retention of the negatively charged amino acid residues, and the differences between the two classification methods were also compared.@*Results@#Among 236 patients, the CALR gene type 1 or type 2 mutation was detected in 206 cases (87.3%), including 173 ET patients (99 cases of type 1 mutation and 74 cases of type 2 mutation) and 33 PMF patients (28 cases of type 1 mutation and 5 cases of type 2 mutation). The CALR non-type 1 or non-type 2 mutation was detected in 30 cases, including 25 ET patients and 5 PMF patients. Among 173 ET patients with CALR gene mutation, the white blood cell count (WBC) of patients with type 1 mutation was higher than that of patients with type 2 mutation [(8.6±2.7)×109/L vs. (7.6±2.4)×109/L, t = 2.45, P = 0.015]. Among 33 PMF patients with CALR gene mutation, the age of patients with type 1 mutation was older than that of patients with type 2 mutation [(58±13) years old vs. (41±16) years old, t = 2.51, P = 0.018]. According to the α-helix propensity score of mutant protein peptide chain and the degree of retention of the negatively charged amino acid residues, 27 kinds of non-type 1 or non-type 2 mutations were classified by using sequencing method, and there were differences between the two methods. According to the α-helix propensity score of the mutant protein peptide chain, the proportion of type 1/type 1-like mutation in PMF patients was higher than that in ET patients [78.9% (30/38) vs. 56.6% (112/198), P < 0.01]. According to the degree of retention of negatively charged amino acid residues in the mutant protein peptide chain, the isoelectric point (pI) value of the mutant protein peptide chain was higher than that of the wild type sequence. The pI value of the type 1-like mutant protein peptide chain was higher than that of the type 2-like mutation (11.79±0.15 vs. 10.02±0.42, t = 11.51, P < 0.01).@*Conclusions@#Type 1 mutated ET patients may be closely related to the high risk of myelofibrosis transformation. The results of the classification of CALR mutations are different according to the α-helix propensity score of the mutant protein peptide chain and the degree of retention of the negatively charged amino acid residues. Further study is necessary to identify the pathogenesis of MPN caused by CALR mutation, and to determine the relationship between mutation type and prognosis of disease.

Kidney injury related to myeloproliferative neoplasms / 中国实用内科杂志

Myeloproliferative neoplasms related kidney injury is clinically rare.It's clinical, histological manifestations are diverse, This article gives a review on the pathogenesis clinical, histological manifestations and current treatments and efficacy, aiming to lay a foundation for the clinical physicians to have a deep understanding of the diagosis and treatment of disease.

Myeloproliferative neoplasms and coronary artery disease / 中国实用内科杂志

Myeloproliferative neoplasms(MPN) are high risk factors of atherosclerosis and arterial thromboembolism. This article gives a review on the incidence, pathogenesis, clinical features and treatment of MPN with coronary artery disease, emphasizing the importance of knowing MPN as possible causes of acute cardiovascular events in patients, in order to help clinicians to learn about the disease and make a proper diagnosis and treatment.