JAK2, CALR, and MPL Mutation Profiles in Colombian patients with BCR-ABL Negative Myeloproliferative Neoplasms

Background: Among the chronic myeloproliferative neoplasms (MPNs) not associated with BCR-ABL mutations are polycythemia vera, primary myelofibrosis, and essential thrombocythemia. These diseases are caused by mutations in genes, such as the JAK2, MPL, and CALR genes, which participate in regulating the JAK-STAT signaling pathway. Objective: This study aimed to establish the frequencies of mutations in the JAK2, MPL, and CALR genes in a group of Colombian patients with a negative clinical diagnosis of BCR-ABL chronic myeloproliferative neoplasms. Methods: The JAK2 V617F and MPL W515K mutations and deletions or insertions in exon 9 of the CALR gene were analyzed in 52 Colombian patients with polycythemia vera, primary myelofibrosis, and essential thrombocythemia. Results: The JAK2V617F mutation was carried by 51.9% of the patients, the CALR mutation by 23%, and the MPL mutation by 3.8%; 23% were triple-negative for the mutations analyzed. In these neoplasms, 6 mutation types in CALR were identified, one of which has not been previously reported. Additionally, one patient presented a double mutation in both the CALR and JAK2 genes. Regarding the hematological results for the mutations, significant differences were found in the hemoglobin level, hematocrit level, and platelet count among the three neoplasms. Conclusion: Thus, this study demonstrates the importance of the molecular characterization of the JAK2, CALR and MPL mutations in Colombian patients (the genetic context of which remains unclear in the abovementioned neoplasms) to achieve an accurate diagnosis, a good prognosis, adequate management, and patient survival.

Antecedentes: Entre las neoplasias mieloproliferativas crónicas no asociadas con mutaciones BCR-ABL se encuentran la policitemia vera, la mielofibrosis primaria y la trombocitemia esencial. Estas enfermedades están causadas por mutaciones en genes, como los genes JAK2, MPL y CALR, que participan en la regulación de la vía de señalización JAK-STAT. Objetivo: Establecer las frecuencias de mutaciones en los genes JAK2, MPL y CALR en un grupo de pacientes colombianos con diagnóstico clínico negativo de NMP BCR-ABL. Metodos: Se analizaron las mutaciones y deleciones o inserciones JAK2 V617F y MPL W515K en el exón 9 del gen CALR en 52 pacientes colombianos con policitemia vera, mielofibrosis primaria y trombocitemia esencial. Resultados: La mutación JAK2V617F la portaban el 51.9% de los pacientes, la mutación CALR el 23.0% y la mutación MPL el 3.8%; El 23.0% fueron triple negativos para las mutaciones analizadas. En estas neoplasias se identificaron seis tipos de mutación en CALR, uno de los cuales no ha sido reportado previamente. Además, un paciente presentó una doble mutación tanto en el gen CALR como en el JAK2. En cuanto a los resultados hematológicos para las mutaciones, se encontraron diferencias significativas en el nivel de hemoglobina, el nivel de hematocrito y el recuento de plaquetas entre las tres neoplasias. Conclusiones: Así, este estudio demuestra la importancia de la caracterización molecular de las mutaciones JAK2, CALR y MPL en pacientes colombianos (cuyo contexto genético aún no está claro en las neoplasias antes mencionadas) para lograr un diagnóstico certero, un buen pronóstico, un manejo adecuado y una mejoría del paciente. supervivencia.

Molecular profile of patients with myelofibrosis: a 10-year experience

ABSTRACT Objective To analyze the karyotype test and myeloid panel with next-generation sequencing findings in patients with myelofibrosis, and to compare transplant characteristics in patients referred for bone marrow transplantation. Methods Retrospective, single-center study with patients diagnosed with myelofibrosis treated at Hospital Israelita Albert Einstein between 2010 and 2020. Results A total of 104 patients with myelofibrosis were examined. Patients who had not been submitted to tests in our service were excluded. The final sample comprised 69 patients. Of these 69, 56 were submitted to karyotyping and 22 to myeloid panel with next-generation sequencing. Karyotype was normal in 60% of the patients and altered in 40%. The prevalence of high-risk molecular mutations was higher in patients referred for bone marrow transplantation (100% versus 50%). The median follow-up of transplant patients was 2.4 years and the overall survival at 2 years was 80% (95%CI: 62-100%). Conclusion The molecular analysis enables estimating the patient's risk and thus instituting more aggressive treatment such as bone marrow transplant for patients at higher risk, being a relevant tool to guide therapy. Given the significance of molecular analysis for therapeutic decision-making in myelofibrosis, collection and disclosure of data on the prevalence of cytogenetic changes and findings of next-generation sequencing in affected patients is important.

Influencia de la carga alélica del JAK2V617F en pacientes cubanos con trombocitemia esencial y mielofibrosis primaria / Influence of JAK2V617F allelic burden in Cuban patients with essential thrombocythemia and primary myelofibrosis

Introducción: La frecuencia de la mutación JAK2V617F se estima entre el 50 y 60 por ciento en pacientes con trombocitemia esencial y mielofibrosis primaria. El 30 por ciento de los pacientes con policitemia vera y mielofibrosis primaria. Entre 2-4 por ciento de los pacientes con trombocitemia esencial presentan pérdida de heterocigosidad. Objetivos: Evaluar la influencia de la carga alélica de la mutación JAK2V617F y su relación con variables clínico-hematológicas en el diagnóstico de estas enfermedades en pacientes cubanos. Métodos: Se realizó un estudio retrospectivo, descriptivo y longitudinal en el Instituto de Hematología e Inmunología entre 2010 y 2020. Se incluyeron todos los pacientes con sospecha de trombocitemia esencial y mielofibrosis primaria con muestras de ADN válidas. Se les cuantificó la carga alélica de la mutación por PCR en tiempo real. Resultados: Se detectó la mutación en 66,7 por ciento de los diagnosticados con trombocitemia esencial y mielofibrosis primaria. El 62,5 por ciento de los pacientes con mielofibrosis primaria fueron homocigotos a la mutación, mientras que en la trombocitemia esencial solo el 20,8 por ciento. La diferencia de medias de cargas alélicas entre ambas enfermedades fue estadísticamente significativa. No se encontraron diferencias significativas en la comparación de las variables clínicas y hematológicas en estas enfermedades ni asociación con la carga alélica con excepción de las plaquetas en la mielofibrosis primaria. Conclusiones: El estudio estuvo limitado por la escasa muestra de pacientes, pero se corresponde con otras investigaciones que sostienen el concepto de que la presentación fenotípica de las neoplasias mieloproliferativasestá influenciada por la carga mutacional del JAK2V617F(AU)

Introduction: The frequency of the JAK2V617F mutation is estimated to be between 50 percent and 60 percent in patients with essential thrombocythemia and primary myelofibrosis. 30 percent of patients with polycythemia vera and primary myelofibrosis and 2-4 percent of patients with essential thrombocythemia show loss of heterozygosity. Objectives: To evaluate the influence of the allelic load of the JAK2V617F mutation in the diagnosis of these diseases in Cuban patients and its relationship with clinical-hematological variables. Methodology: A retrospective, descriptive and longitudinal study was carried out at the Institute of Hematology and Immunology between 2010 and 2020. All patients with suspected essential thrombocythemia and primary myelofibrosis with valid DNA samples were included. The allelic load of the mutation was quantified by real-time PCR. Results: The mutation was detected in 66.7 percent of those diagnosed with essential thrombocythemia and primary myelofibrosis. 62.5 percent of the patients with primary myelofibrosis were homozygous for the mutation, while in essential thrombocythemia only 20.8 percent. The difference in mean allelic loads between both diseases was statistically significant. No significant differences were found in the comparison of clinical and hematological variables in these diseases or association with allelic load, with the exception of platelets in primary myelofibrosis. Conclusions: The study was limited by the small sample of patients, but it corresponds to other investigations that support the concept that the phenotypic presentation of myeloproliferative neoplasms is influenced by the mutational load of JAK2V617F(AU)

Transformation of primary myelofibrosis to B-cell acute lymphoblastic leukemia: report of 1 case and review of literature / 白血病·淋巴瘤

Objective:To investigate the pathogenesis and prognosis of transformation of primary myelofibrosis (PMF) to B-cell acute lymphoblastic leukemia (B-ALL).Methods:The diagnosis and treatment process of a patient transferred from PMF to B-ALL in Affiliated Tumor Hospital of Zhengzhou University in November 2018 were retrospectively analyzed, and the relevant literature was reviewed.Results:The patient was a 64-year-old female, she was initially diagnosed with PMF, and then she developed B-ALL 17 months later after receiving treatment of prednisone, danazole, levamisole, aspirin, thalidomide and jaktinib. After induction therapy, the patient received 8 months of continuous remission, and then the reexamination showed relapse. There was no remission after reinduction therapy. The patient gave up treatment and was discharged 2 months later. JAK2 V617F gene mutation was positive before and after leukemia transformation.Conclusions:The patients with transformation of PMF to B-ALL have poor clinical prognosis and short survival time. The possible mechanism of its transformation may be related to additional genetic events or certain high-risk genes. However, the specific mechanism is still unclear, and further investigation of the etiology is needed to seek targeted treatment.

Características clínicas y epidemiológicas de las neoplasias mieloproliferativas Philadelphia negativas en el sistema público de salud de Chile

Background: Philadelphia-negative myeloproliferative neoplasms (Ph-MPN) are chronic hematological disorders characterized by the overproduction of one or more mature myeloid blood cell lineages. Classical Ph-MPN are polycythemia vera (PV), essential thrombocytopenia (ET) and primary myelofibrosis (PMF). Aim: To assess the epidemiological, clinical and diagnostic characteristics of Ph-MPN in Chile. Material and Methods: Retrospective review of medical records of all patients referred as MPN from 2012 to 2017. Patients with (9;21) translocation were excluded. Results: Data of 462 cases with a median age of 69 years from 10 public hospitals was reviewed. ET was the most frequently Ph-MNP found. The incidence of Ph-MPN was 1.5 x 100.000 cases. The JAK2 V617F mutation study was performed in 96% of patients and only 30% had a bone marrow biopsy. Thrombotic events were observed in 29% of patients. Bleeding events were observed in 7%. Five-year overall survival was 87%. Conclusions: ET is the most frequent Ph-MPN. The mean incidence was lower than reported in the literature, in part because of a sub diagnosis.

Clinical features and prognosis of primary myelofibrosis with hepatic vascular disease and/or portal hypertension / 临床肝胆病杂志

Objective To investigate the clinical features, liver histopathological features, treatment, and prognosis of primary myelofibrosis (PMF)-associated hepatic vascular disease and portal hypertension. Methods A retrospective analysis was performed for the clinical and pathological features of 68 patients who were diagnosed with PMF in 960 Hospital of the PLA Joint Logistics Support Force and Xijing Hospital of Digestive Diseases, Air Force Medical University, from July 2010 to December 2020, among whom 22 patients had hepatic vascular disease/portal hypertension as the main manifestation. The patients were divided into two groups according to the presence or absence of thrombosis, and treatment and prognosis were summarized. The Kaplan-Meier method was used to plot survival curves, and the log-rank test was used to compare long-term survival rate between the two groups. Results Among the 68 patients with PMF, 22 had hepatic vascular disease and/or portal hypertension, resulting in a prevalence rate of 32.35%, and among these 22 patients, 13 (59.1%) had extrahepatic portal vein occlusion, 1 (4.5%) had Budd-Chiari syndrome, and 8 (36.4%) had portal hypertension. Biopsy was performed for 7 patients, and pathological results showed extramedullary hematopoiesis in the liver and varying degrees of infiltration of lymphocytes, plasma cells, and eosinophils at the lobular and portal areas, but the lobular structure was normal. A total of 7 patients died during follow-up, among whom 5 died of complications associated with thrombosis or portal hypertension. The overall median survival time was 57.99 months for all patients; the median survival time was 45.33 months in patients with thrombosis and 64.00 months in patients without thrombosis, and although there was no significant difference between the two groups ( χ 2 =3.035, P =0.081), the non-thrombosis group tended to have better survival and prognosis than the thrombosis group. Conclusion The possibility of PMF as the primary disease should be considered for patients with hepatic vascular disease and portal hypertension. Patients with PMF should be screened for hepatic vascular disease, and early intervention should be given. The patients without thrombosis tend to have better survival and prognosis than those with thrombosis.

Meta-analysis of the frequency of JAK2 in primary myelofibrosis according to the detection method used / Metaanálisis de la frecuencia de JAK2 en mielofibrosis primaria según la técnica de detección

Abstract Background and objective: in the global scientific literature, the frequency of JAK2 is highly heterogenous in chronic myeloproliferative neoplasms. The objective of this study was to analyze the prevalence of the JAK2 mutation in primary myelofibrosis (PMF) and compare it according to the detection method used, from 2007-2018. Materials and methods: a systematic review with meta-analysis, using 21 searches in three multidisciplinary databases. The PRISMA guideline phases of identification, screening, selection and inclusion were applied. Reproducibility and evaluation of the methodological quality were ensured. The analyses were based on frequencies and meta-analysis for the prevalence of the mutation with its 95% confidence interval. Results: twenty-nine studies with 744 patients were included, mainly from Korea, Brazil and China. The most commonly used technique was AS-PCR, and the prevalence of JAK2 with this technique ranged from 33.3 to 71.4%; with real-time PCR ranging from 42.9 to 77.3%, sequencing from 14.3-57.4%, and ARMS from 36.4-83.3%. The prevalence of JAK2 showed no statistically significant differences according to the type of diagnostic test used. Conclusion: high frequencies of the JAK2V617F mutation are seen in PMF, which shows that this entity should not be diagnosed solely based on clinical and hematological characteristics, but also on the patients' genetic screening.

Resumen Antecedentes y objetivo: en la literatura científica mundial la frecuencia de JAK2 presenta una alta heterogeneidad en las neoplasias mieloproliferativas crónicas. El objetivo de esta investigación fue analizar la prevalencia de la mutación JAK2 en mielofibrosis Primaria (MFP) y compararla según la técnica de detección en los años 2007-2018. Material y métodos: revisión sistemática con metaanálisis, usando 21 búsquedas en tres bases de datos multidisciplinarias. Se aplicaron las fases de identificación, tamización, elección e inclusión de la guía PRISMA. Se garantizó reproducibilidad y evaluación de la calidad metodológica. Los análisis se basaron en frecuencias y metaanálisis para la prevalencia de la mutación con su intervalo de confianza de 95%. Resultados: se incluyeron 29 estudios con 744 pacientes, los cuales provienen principalmente de Corea, Brasil y China. La técnica más empleada fue AS-PCR, las prevalencias de JAK2 con esta técnica oscilaron entre 33.3 y 71.4%; con PCR en tiempo real entre 42.9 y 77.3%, con secuenciación de 14.3-57.4% y en ARMS de 36.4-83.3%. La prevalencia de JAK2 no presentó diferencias estadísticamente significativas según el tipo de prueba diagnóstica utilizada. Conclusión: se evidencian altas frecuencias de la mutación JAK2V617F en MFP, lo que evidencia que el diagnóstico de esta entidad no debe realizarse únicamente por características clínicas y hematológicas sino también por la tamización genética de los pacientes.

Clinical and molecular profile of a Brazilian cohort of patients with classical BCR-ABL1-negative myeloproliferative neoplasms

ABSTRACT Background: The classical BCR-ABL1-negative myeloproliferative neoplasms (MPNs) are Polycythemia Vera (PV), Essential Thrombocythemia (ET) and Primary Myelofibrosis (PMF). In developing countries, there are few reports that truly reveal the clinical setting of these patients. Therefore, we aimed to characterize a single center MPN population with a special focus on the correct diagnosis based on the recent review of the WHO criteria for the diagnosis of myeloid neoplasms. Methods: This retrospective study analyzed data from medical records of patients with classical BCR-ABL1-negative MPNs diagnosed from January 1997 to October 2017 and followed at the University Hospital of Ribeirão Preto Medical School. Results: A total of 162 patients were assessed, 61 with PV, 50 with ET, and 51 with PMF. The mutational status analysis revealed that 113 (69.3%) harbored the JAK2V617F mutation, 23 (14.1%), the CALR mutation, and 12 (7.4%) had a triple-negative status. None of the patients were found to have mutations on the thrombopoietin receptor gene (MPL), including some ET and PMF patients who were not tested. Among the PV patients, 57 (93.5%) were positive for the JAK2V617F mutation, one (1.6%) presented an in-frame deletion JAK2 exon 12 mutation and one (1.6%) presented a missense JAK2 exon 9 mutation, not previously described. The overall survival was lower in the triple-negative patients with PMF, when compared to the JAK2V617F or CALR-mutated (p= 0.002). Conclusion: The frequency of somatic mutations and survival in our cohort, stratified according to the respective disease, was consistent with the literature data, despite some limitations. Further prospective epidemiological studies of MPN cohorts are encouraged in developing countries.

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).

Ruxolitinib combined with prednisone, thalidomide and danazol for treatment of myelofibrosis: a pilot study / 中华血液学杂志

Objective@#To evaluate the efficacy and tolerability of ruxolitinib combined with prednisone, thalidomide and danazol for treatment of in myelofibrosis (MF).@*Methods@#Patients of MF according to the WHO 2016 criteria, received ruxolitinib (RUX) combined with prednisone, thalidomide and danazol (PTD). The response, changes of blood counts and adverse events were evaluated.@*Results@#Six PMF and one post-ET MF patients were enrolled. Four patients presented JAK2V617F mutation, one CALR mutation, one MPL mutation, one triple-negative. Responses per IWG-MRT criteria were clinical improvement in 5 patients, stable disease in 2 ones, spleen response in 6 ones. All of 7 patients were symptomatic responses, four patients achieved at least 50% improvement from baseline on MPN-SAF TSS. Three patients initially treated with RUX alone, all of 3 patients experienced treatment-associated anemia and thrombocytopenia. Then these 3 patients received RUX combined with PTD, both hemoglobin and platelet increased significantly. Four patients initially treated with RUX combined with PTD. Increased levels of hemoglobin and platelet were seen in all of 7 patients received RUX combined with PTD with maximum increased hemoglobin of 30(18-54) g/L and maximum increased platelets of 116(13-369)×109/L, respectively from baseline. The treatment dose of RUX increased due to improved platelet count in 3 patients. The frequent non-hematologic adverse events grade 1-2 were constipation, abdominal distension, crura edema and increased ALT.@*Conclusions@#RUX combined with PTD for treatment of MF may modulate initial hematologic toxicity observed when RUX alone, and may increase response due to improved levels of hemoglobin or platelet.

A study of clinical characteristics and prognosis of primary myelofibrosis patients with thrombocytopenia in varied degrees / 中华血液学杂志

Objective@#To evaluate clinical characteristics and prognosis of primary myelofibrosis (PMF) patients with thrombocytopenia in varied degrees.@*Methods@#Clinical features and survival data of 1 305 Chinese patients with PMF were retrospectively analyzed. The prognostic value of thrombocytopenia in patients with PMF was evaluated.@*Results@#320 subjects (47%) presented severe thrombocytopenia (PLT<50×109/L), 198 ones (15.2%) mild thrombocytopenia [PLT (50-99)×109/L] and 787 ones (60.3%) without thrombocytopenia (PLT ≥ 100×109/L). The more severe the thrombocytopenia, the higher the proportions of HGB<100 g/L, WBC<4×109/L, circulating blasts ≥ 3%, abnormal karyotype and unfavourable cytogenetics (P<0.001, P<0.001, P=0.004, P<0.001 and P<0.001, respectively) were observed in this cohort of patients. The more severe the thrombocytopenia, the lower the proportion of JAK2V617F positive (P<0.001) was also noticed. Platelet count was positively correlated with splenomegaly, HGB and WBC (P<0.001, correlation coefficients were 0.131, 0.445 and 0.156, respectively). Platelet count was negative correlated with constitutional symptoms and circulating blasts (P=0.009, P=0.045, respectively; correlation coefficients were -0.096 and -0.056, respectively). The median survival of patients with severe thrombocytopenia, mild thrombocytopenia and without thrombocytopenia were 32, 67 and 89 months, respectively (P<0.001). Multivariate analysis identified thrombocytopenia in varied degrees (HR=1.693, 95%CI 1.320-2.173, P<0.001) and Dynamic Internation Prognostic Scoring System(DIPSS) prognostic model (HR=2.051, 95%CI 1.511-2.784, P<0.001) as independent risk factors for survival.@*Conclusion@#PMF patients with severe thrombocytopenia frequently displayed anemia, leucopenia, circulating blasts and short survival, so active treatment measures should be taken especially in these patients.

Symptom load assessment of Ph chromosome/BCR-ABL fusion gene negative myeloproliferative tumor patients at different efficacy evaluation periods / 国际肿瘤学杂志

Objective@#To observe the disease type and the changes of symptom load during treatment of patients with Ph chromosome/BCR-ABL fusion gene negative myeloproliferative neoplasm (MPN).@*Methods@#A total of 84 patients with MPN diagnosed from May 2017 to January 2019 in People′s Hospital of Longhua District of Shenzhen were selected, and were divided into polycythemia vera (PV) group, essential thrombocyhemia (ET) group, and myelofibrosis (PMF) group according to their subtypes, with 28 cases in each group. The scores of MPN-SAF-TSS were compared among the three groups. Besides, the scores of the scale (myeloproliferative neoplasm symptom assessment form total symptom score, MPN-SAF-TSS) in different treatment periods (at the time of the visit, when the disease progressed, when the disease was stable, when the clinical improvement was made, when the partial remission was completed, at the time of remission and recurrence) were also compared.@*Results@#At the time of initial diagnosis, there were significant differences in the incidences of symptom burdens among the three groups of MPN patients with abdominal fullness (χ2=6.095, P=0.047), abdominal discomfort (χ2=7.342, P=0.025), poor mobility (χ2=13.029, P=0.001), inattention (χ2=6.099, P=0.047), pruritus (χ2=6.956, P=0.031), bone pain (χ2=7.807, P=0.020), fever (χ2=8.000, P=0.018) and weight loss (χ2=27.340, P<0.001). The incidences of poor mobility (85.71%, 24/28), inattention (67.86%, 19/28) and weight loss (82.14%, 23/28) in PMF group were significantly higher than those in PV group [42.86% (12/28), 39.29% (11/28), 35.71% (10/28)] and ET group [46.43% (13/28), 39.29% (11/28), 14.29% (4/28)] (all P<0.05). The incidences of abdominal discomfort (75.00%, 21/28) and bone pain (60.71%, 17/28) in PMF group were higher than those in PV group [39.29% (11/28), 25.00% (7/28)] (both P<0.05). The incidences of abdominal fullness (89.29%, 25/28) and fever (42.86%, 12/28) in PMF group were higher than those in ET group [60.71% (17/28), 10.71% (3/28)] (both P<0.05). The incidence of pruritus in PV group (71.43%, 20/28) was higher than that in ET group (42.86%, 12/28) and PMF group (39.29%, 11/28) (both P<0.05). Symptom load scores of patients with fatigue (χ2=368.594, P<0.001), abdominal fullness (χ2=261.312, P<0.001), abdominal discomfort (χ2=195.629, P<0.001), poor mobility (χ2=217.862, P<0.001), lack of concentration (χ2=280.664, P<0.001), night sweats (χ2=239.650, P<0.001), pruritus (χ2=254.418, P<0.001), bone pain (χ2=180.291, P<0.001), fever (χ2=231.613, P<0.001) and weight loss (χ2=227.831, P<0.001) were significantly different during different therapeutic periods. The fatigue symptom load score was higher when the disease progressed than that at the time of the visit (P<0.05), and the symptom score of abdominal fullness was lower than that at the time of visit (P<0.05). Symptom load scores of weakness and pruritus when the condition was stable was lower than those when the disease progressed (both P<0.05). When the clinical improvement was made, symptom load scores of weakness, abdominal discomfort, inattention, night sweats, weight loss were lower than those when the disease was stable (all P<0.05). Symptom load scores of abdominal fullness, poor mobility, inattention, night sweats and pruritus in partial remission period decreased compared to temporary improvement period (all P<0.05). Compared to the partial remission period, the symptom load scores of weakness, abdominal fullness, night sweats, pruritus, bone pain and weight loss in complete remission period were lower (all P<0.05). At last, symptom load scores of weakness, abdominal fullness, abdominal discomfort, poor mobility, inattention, night sweats, pruritus, bone pain, fever and weight loss in recurrence period were higher than those in complete remission period (all P<0.05).@*Conclusion@#There are several differences in the main clinical symptoms among patients with different MPN subtypes, and there are significant changes in the main clinical symptoms as the disease progresses or turns around.

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.

Symptom load assessment of Ph chromosome/ BCR-ABL fusion gene negative myeloproliferative tumor patients at different efficacy evaluation periods / 国际肿瘤学杂志

Objective To observe the disease type and the changes of symptom load during treatment of patients with Ph chromosome/ BCR-ABL fusion gene negative myeloproliferative neoplasm (MPN). Methods A total of 84 patients with MPN diagnosed from May 2017 to January 2019 in People's Hospital of Longhua Dis-trict of Shenzhen were selected,and were divided into polycythemia vera (PV)group,essential thrombocy-hemia (ET)group,and myelofibrosis (PMF)group according to their subtypes,with 28 cases in each group. The scores of MPN-SAF-TSS were compared among the three groups. Besides,the scores of the scale (myelo-proliferative neoplasm symptom assessment form total symptom score,MPN-SAF-TSS)in different treatment periods (at the time of the visit,when the disease progressed,when the disease was stable,when the clinical improvement was made,when the partial remission was completed,at the time of remission and recurrence) were also compared. Results At the time of initial diagnosis,there were significant differences in the inci-dences of symptom burdens among the three groups of MPN patients with abdominal fullness (χ2 = 6. 095,P =0. 047),abdominal discomfort (χ2 = 7. 342,P = 0. 025),poor mobility (χ2 = 13. 029,P = 0. 001),inatten-tion (χ2 = 6. 099,P = 0. 047),pruritus (χ2 = 6. 956,P = 0. 031),bone pain (χ2 = 7. 807,P = 0. 020),fever (χ2 = 8. 000,P = 0. 018)and weight loss (χ2 = 27. 340,P < 0. 001). The incidences of poor mobility (85. 71％,24 / 28),inattention (67. 86％,19 / 28)and weight loss (82. 14％,23 / 28)in PMF group were significantly higher than those in PV group [42. 86％ (12 / 28),39. 29％ (11 / 28),35. 71％ (10 / 28)]and ET group [46. 43％ (13 / 28),39. 29％ (11 / 28),14. 29％ (4 / 28)](all P < 0. 05). The incidences of abdominal discomfort (75. 00％,21 / 28)and bone pain (60. 71％,17 / 28)in PMF group were higher than those in PV group [39. 29％ (11 / 28),25. 00％ (7 / 28)](both P < 0. 05). The incidences of abdominal fullness (89. 29％,25 / 28)and fever (42. 86％,12 / 28)in PMF group were higher than those in ET group [60. 71％ (17 / 28),10. 71％ (3 / 28)](both P < 0. 05). The incidence of pruritus in PV group (71. 43％, 20 / 28)was higher than that in ET group (42. 86％,12 / 28)and PMF group (39. 29％,11 / 28)(both P <0. 05). Symptom load scores of patients with fatigue (χ2 = 368. 594,P < 0. 001),abdominal fullness (χ2 =261. 312,P < 0. 001),abdominal discomfort (χ2 = 195. 629,P < 0. 001),poor mobility (χ2 = 217. 862,P <0. 001),lack of concentration (χ2 = 280. 664,P < 0. 001),night sweats (χ2 = 239. 650,P < 0. 001),pruri-tus (χ2 = 254. 418,P < 0. 001),bone pain (χ2 = 180. 291,P < 0. 001),fever (χ2 = 231. 613,P < 0. 001) and weight loss (χ2 = 227. 831,P < 0. 001)were significantly different during different therapeutic periods. The fatigue symptom load score was higher when the disease progressed than that at the time of the visit (P <0. 05),and the symptom score of abdominal fullness was lower than that at the time of visit (P < 0. 05). Symp-tom load scores of weakness and pruritus when the condition was stable was lower than those when the disease progressed (both P < 0. 05). When the clinical improvement was made,symptom load scores of weakness, abdominal discomfort,inattention,night sweats,weight loss were lower than those when the disease was stable (all P < 0. 05). Symptom load scores of abdominal fullness,poor mobility,inattention,night sweats and pruri-tus in partial remission period decreased compared to temporary improvement period (all P < 0. 05). Compared to the partial remission period,the symptom load scores of weakness,abdominal fullness,night sweats,pruri-tus,bone pain and weight loss in complete remission period were lower (all P < 0. 05). At last,symptom load scores of weakness,abdominal fullness,abdominal discomfort,poor mobility,inattention,night sweats,pruri-tus,bone pain,fever and weight loss in recurrence period were higher than those in complete remission period (all P < 0. 05). Conclusion There are several differences in the main clinical symptoms among patients with different MPN subtypes,and there are significant changes in the main clinical symptoms as the disease progresses or turns around.

CD47 expression is decreased in hematopoietic progenitor cells in patients with myelofibrosis

Myelofibrosis (MF) is characterized by increased circulating hematopoietic progenitor cells (HPCs), abnormal cytokine levels, and the survival advantage of neoplastic progenitors over their normal counterparts, which leads to progressive disappearance of polyclonal hematopoiesis. CD47 is a surface glycoprotein with many functions, such as acting as a phagocytosis inhibitor of the expressing cell, that is increased in normal hematopoietic stem and progenitor cells mobilized into the blood and several human cancer-initiating cells, such as in acute myeloid leukemia. We compared CD47 expression in hematopoietic stem and progenitor cells of patients with MF and controls and found it to be decreased in progenitors of MF. Exposure of control HPCs to the cytokines transforming growth factor β and stromal-derived factor 1, which are important regulators of hematopoietic stem cell cycling and are overexpressed in patients with MF, did not modulate CD47 expression.

Male with myelofibrosis and ulceronecrotic lesions

SUMMARY Granulocytic sarcoma also called myeloid sarcoma is an extramedullary tumour of immature granulocytic cells. It is a rare entity, and mostly accompanied by acute myeloid leukaemia. It is observed during the course of myeloproliferative disorders especially in chronic myeloid leukaemia and myelodysplastic syndromes. Here, we report a case of a 60-year-old male with past history of myelofibrosis admitted to the emergency room due ulceronecrotic lesions, fever and dysphagia. We emphasize the importance of recognizing this entity and its severity.

RESUMO O sarcoma granulocítico, também chamado de sarcoma mieloide, é um tumor extramedular de células granulocíticas ¡maturas. É uma entidade rara, e principalmente acompanhada de leucemia mieloide aguda. É observado durante o transtorno mieloproliferativo, especialmente na leucemia mieloide crónica e síndromes mielodisplásicas. Aqui, relatamos um caso de um homem de 60 anos com antecedente de mielofibrose admitida na sala de emergência devido a lesões ulceronecróticas, febre e disfagia. Enfatizamos a importância de reconhecer essa entidade e sua gravidade.

Mielofibrosis primaria manifestada como fiebre de origen desconocido / Primary myelofibrosis manifested as fever of unknown origin

Resumen Se comunica el caso de un paciente de 55 años de edad que inició con fiebre como síntoma principal de su padecimiento. Después de un protocolo de estudio sistematizado y orientado se llegó al diagnóstico de fiebre de origen desconocido como síndrome, cuyo diagnóstico etiológico fue mielofibrosis primaria, enfermedad hematológica poco frecuente que se distingue por fibrosis de la médula ósea, esplenomegalia y anemia. Se revisa el abordaje de la fiebre y sus etapas de estudio siguiendo las recomendaciones actuales, técnicas y científicas, pero adaptadas a nuestro medio y recursos.

Abstract This paper reports the case of a 55 year-old man who presented with fever as a primary symptom of his condition. A systematic protocol oriented study led to the diagnosis of fever of unknown origin as a syndrome whose etiologic diagnosis was primary myelofibrosis; rare hematologic disease characterized by fibrosis of the bone marrow, anemia and splenomegaly. A review of the approach of fever and stages of study is done according to current recommendations, technical and scientific, but adapted to our environment and resources.

Perfil dos pacientes com Neoplasia Mieloproliferativa Cromossomo Philadelfia Negativo na Unidade de Alta Complexidade do Hospital São José em Criciúma/SC no período de 2008 a 2015 / Profile of patients with Philadelfia Negative Myeloproliferative neoplasm in São José Hospital's High Complexity Oncologic Unit in Criciúma/SC in the 2008 to 2015 period

O presente artigo tem como objetivo conhecer o perfil dos pacientes portadores de Neoplasias Mieloproliferativas Crônicas (NMPC) cromossomo Philadelfia (Ph) negativo clássicas que compreendem as entidades Policitemia Vera (PV), Trombocitose Essencial (TE) e Mielofibrose Primária (MP) na Unidade de Alta Complexidade Oncológica no período de 2008 a 2015. Trata-se de um estudo observacional, descritivo, retrospectivo e de abordagem quantitativa em que foram avaliados 77 prontuários de pacientes com diagnóstico de Policitemia Vera, Trombocitose Essencial e Mielofibrose Primária. Os resultados demonstraram que 54,5% (n=42) tinham diagnóstico de Trombocitose Essencial, 27,3% (n=21) de Policitemia Vera e 18,2% (n=14) de Mielofibrose Primária. A idade média ao diagnóstico girou em torno da sexta e sétima década de vida. As principais alterações laboratoriais encontradas na Policitemia Vera e na Mielofibrose Primária foram a hemoglobina, a qual tem como média 16,05(±2,89) e 9,98(±3,31) respectivamente, e o hematócrito com média de 51,48(±8,69) e 30,70 (±10,14) nesta ordem; já na Trombocitose Essencial as plaquetas foram a principal alteração, com média de 1.728.000(±447136,4). Em relação aos eventos tromboembólicos, 28,6% dos pacientes com PV, 21,4% com TE e 14,3% com MI apresentaram algum tipo de evento (IAM, TVP ou AVC).A mutação JAK2 foi positiva principalmente em portadores de Policitemia Vera, 76,2% (n=16). Dos pacientes portadores de Policitemia Vera 85,7% (n=18), 95,2% (n=40) de Trombocitose Essencial e 57,1% (n=8) de MP permanecem vivos e em tratamento. Concluise que o sexo feminino foi o mais acometido, a idade média ao diagnóstico foi entre a sexta e sétima década de vida, sendo a Trombocitose Essencial a doença mais incidente.

The present article aims to know the profile of patients with classic Philadelphia-negative Chronic Myeloproliferative Neoplasms (CMPNs) who comprehend the entities Polycythemia Vera (PV), Essential Thrombocytosis (ET) and Primary Myelofibrosis (PM) at the High Complexity Oncology Unit between 2008 and 2015. This is an observational, descriptive, retrospective and of quantitative approach study in which 77 diagnosed with Polycythemia Vera, Essential Thrombocytosis and Primary Myelofibrosis patients' charts were analyzed. Results demonstrated that 54,5% (n=42) were diagnosed with Essential Thrombocytosis, 27,3% (n=21) with Polycythemia Vera and 18,2% (n=14) with Primay Myelofibrosis. The mean age at diagnosis was around the sixth and seventh decade of life. The main laboratorial alterations found in Polycythemia Vera and Primary Myelofibrosis were hemoglobin, which has mean 16,05(±2,89) and 9,98(±3,31), respectively, and hematocrit, with mean 51,48(±8,69) and 30,70 (±10,14), in that order; in Essential Thrombocytosis, platelets were the main alteration, with mean 1.728.000(±447136,4). The frequency of thrombotic event was 64,3%, most of which had the diagnosis of Essential Thrombocytosis. The JAK2 mutation was positive mainly on patients with Polycythemia Vera, 76,2% (n=16). 85,7% (n=18) Polycythemia Vera, 95,2% (n=40) Essential Thrombocytosis and 57,1% (n=8) Primary Myelofibrosis patients remain alive and on treatment. It was concluded that females were the most affected, that the mean age at diagnosis was between the sixth and seventh decade of life and that Essential Thrombocytosis was the most incident disease.

Progress of treatment of primary myelofibrosis / 白血病·淋巴瘤

Primary myelofibrosis (PMF) is a type of myeloproliferative neoplasms (MPN) with the worst prognosis in MPN. Immunomodulators, erythropoiesis-stimulating agents, cytoreductive drugs, splenectomy, etc. can significantly reduce the symptoms of patients, but can not improve the prognosis of patients. Allogeneic hematopoietic stem cell transplantation remains the only method that may cure PMF. The marketed JAK kinase inhibitor ruxolitinib and the hypomethylating agents, anti-fibrosis agents and telomerase inhibitorprovide new ideas for the treatment of PMF, made it possible to reverse the natural course of myelofibrosis. This paper reviews the progress of PMF therapy.

Primary Myelofibrosis with MPL S505N Mutation: The First Case Reported in Korea

MPL mutation is an important molecular marker in myeloproliferative neoplasms (MPN). Although MPL W515 is a hot spot for missense mutations in MPN, MPL S505 mutations have been reported in both familial and non-familial MPN. A 72-year-old male visited the hospital, complaining mainly of dizziness and epistaxis. Leukocytosis, anemia, thrombocytopenia, tear drop cells, nucleated RBCs, and myeloblasts were observed in both complete blood cell counts and peripheral blood smears. Bone marrow aspiration failed due to dilution with peripheral blood. BM biopsy indicated hypercellular marrow, megakaryocytic proliferation with atypia, and grade 3 reticulin fibrosis. Conventional cytogenetics results were as follows: 46,XY,del(13)(q12q22)[19]/46,XY[1]. Molecular studies did not detect JAK2 V617F, BCR/ABL translocation, JAK2 exon 12, and CALR exon 9 mutations. The MPL S505N mutation was verified by colony PCR and Sanger sequencing following gene cloning. Based on the above findings, a diagnosis of overt primary myelofibrosis (PMF) was indicated. Mutation studies of buccal and T cells were not conducted. Further, family members were not subjected to mutation studies. Therefore, we were unable to determine whether this mutation was familial or non-familial. Six months after the first visit to the hospital, the patient died due to pneumonia and sepsis. Thrombotic symptoms or major bleeding events did not develop during the survival period following diagnosis of PMF. To the best of our knowledge, this may be the first reported case of PMF with the MPL S505N mutation in Korea.