scholarly journals Survival in Essential Thrombocythemia and Prefibrotic Primary Myelofibrosis - Correlation of Clinical Phenotype with Histomorphological Diagnosis

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 1932-1932
Author(s):  
Georg Jeryczynski ◽  
Albert Woelfler ◽  
Bettina Gisslinger ◽  
Martin Schalling ◽  
Sonja Burgstaller ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Overall Survival ◽  
Clinical Features ◽  
Correct Diagnosis ◽  
Molecular Genetic ◽  
Myeloproliferative Neoplasm ◽  
Primary Myelofibrosis ◽  
Diagnostic Systems ◽  
Bone Marrow Histology ◽  
Survival Patterns

Abstract Introduction The correct diagnosis of myeloproliferative neoplasm (MPN), especially the correct classification of early or prodromal stages, remains a matter of debate. While the 2008 WHOcriteriarecognize the existence of an early, prefibrotic primary myelofibrosis, other diagnostic systems make no such distinction between ET and prePMF. According to WHO criteria the accurate diagnosis is based mainly onhistomorphologicassessment of bone marrow (BM) biopsy samples. However, there is an ongoing debate if ET, prePMF, PV and overt PMF are, dependent from their molecular genetic phenotype, a continuum or rather independent entities. Methods From a multicenter database, we selected 251 patients with ET and 185 patients with prePMF, in which the diagnosis had been established by BM biopsy and in which clinical data at time of diagnosis and for follow up were available. We compared overall survival in patients from both entities in which common clinical features of prePMF were present at time of bone marrow biopsy. These were anemia (male <13 g/dL, female <12 g/dL),leukocyte counts ³ 11 G/L, elevated lactate dehydrogenase levels (LDH) as well as splenomegaly. Results Survival was substantially impaired in WHO-defined prePMF patients when compared to the ET cohort (Fig. 1). Median overall survival was significantly worse in patients with leukocytosis (9.3 vs. 14.9 years, 95% CI 6.9-11.6 vs. 9.1-20.7, p<0.001), elevated LDH levels (10.6 vs. 21.7 years, 95% CI 7.7-13.4 vs. 11.5-31.9, p<0.001) and splenomegaly (6.8 years vs. median not reached, 95% CI 6.8-10.8 vs.n.a.,p<0.001). Significance was narrowly missed in anemic patients (7.2 vs. 13.1 years, 95% CI 4.7-9.8 vs. 9.2-16.9, p=0.089). Discussion Our data illustrate, that the distinction of prePMF and ET patients based onhistomorphologicalcriteria by the WHO 2008 diagnostic criteria translates into different overall survival patterns in patients that may share some similar clinical features at diagnosis. This suggests that the underlying diseases are two different entities, with different underlying biology, that can both present with the same adverse clinical parameters. Further, these two entities can only be accurately differentiated by bone marrow histology. Therefore the concept that ET, WHO-defined prePMF and overt PMF form a continuum of the same disease, with prePMF just being an advanced stage of ET, may need revision in face of this study. Figure 1 Figure 1. Disclosures Burgstaller: Novartis: Consultancy, Honoraria. Geissler:Novartis: Honoraria. Gisslinger:AOP Orphan: Consultancy, Honoraria; Novartis: Consultancy, Honoraria; Baxalta: Consultancy, Honoraria.

scholarly journals Focus on Osteosclerotic Progression in Primary Myelofibrosis

Biomolecules ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 122
Author(s):  
Mariarita Spampinato ◽  
Cesarina Giallongo ◽  
Alessandra Romano ◽  
Lucia Longhitano ◽  
Enrico La Spina ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Marrow Cell ◽  
Myeloproliferative Neoplasm ◽  
Primary Myelofibrosis ◽  
Hematopoietic Stem ◽  
Clonal Proliferation ◽  
Cytokines And Chemokines ◽  
Bone Damage ◽  
Underlying Mechanisms ◽  
Modulating Functions

Primary myelofibrosis (PMF) is a myeloproliferative neoplasm characterized by hematopoietic stem-cell-derived clonal proliferation, leading to bone marrow (BM) fibrosis. Hematopoiesis alterations are closely associated with modifications of the BM microenvironment, characterized by defective interactions between vascular and endosteal niches. As such, neoangiogenesis, megakaryocytes hyperplasia and extensive bone marrow fibrosis, followed by osteosclerosis and bone damage, are the most relevant consequences of PMF. Moreover, bone tissue deposition, together with progressive fibrosis, represents crucial mechanisms of disabilities in patients. Although the underlying mechanisms of bone damage observed in PMF are still unclear, the involvement of cytokines, growth factors and bone marrow microenvironment resident cells have been linked to disease progression. Herein, we focused on the role of megakaryocytes and their alterations, associated with cytokines and chemokines release, in modulating functions of most of the bone marrow cell populations and in creating a complex network where impaired signaling strongly contributes to progression and disabilities.

Clinical Features of Chinese Refractory Cytopenia of Childhood: Lower Overall Survival and Higher Possibillity of Clonal Evolution Than Nsaa Patients

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4947-4947
Author(s):  
Wenbin An ◽  
Zhanqi Li ◽  
Shuchun Wang ◽  
Xiaojuan Chen ◽  
Wenyu Yang ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Overall Survival ◽  
Survival Time ◽  
Clinical Features ◽  
Bone Marrow Biopsy ◽  
Lymphocyte Count ◽  
Clonal Evolution ◽  
Scoring Model ◽  
Erythroblastic Island

Abstract Abstract 4947 Objective: To analyze the clinical features and survival time in patients with refractory cytopenia of childhood (RCC), and to develop a new diagnostic scoring system for RCC. Methods: The clinical information, laboratory findings at primary diagnosis and survival time in 97 non-severe aplastic anemia (NSAA) children diagnosed in August, 2000 to June, 2006 were analyzed retrospectively. According to the criteria for RCC proposed in the 2008 edition of the World Health Organization classification of hematopoietic and lymphoid tissues, diagnosis of these NSAA patients was reassessed. Results: 37/97 cases (38. 1%) were reassessed as RCC, 60/97 cases (61. 9%) were still diagnosed as NSAA. There was no significant differences in distribution of age, gender and chromosomal abnormalities, absolute neutrophil count, PLT, MCV, absolute reticulocyte count, the number of cytopenias and duration from onset to starting treatmeat between RCC and NSAA. All patients received the treatment of CsA and androgen. There was no significant differences in distribution of treatment response. The overall survival (OS) of RCC patients was significantly lower than NSAA patients (P=0. 041) in long-term follow-up. The former had 91. 5% of 6-year prospective survival (PS) and the latter had 96. 5% of 6-year PS. At a median follow-up of 93 months (range 6–138), 2 patients reassessed as RCC progressed into AML-M5 and PNH respectively, while no NSAA patient progressed into clonal diseases. There was significant differences in the distribution of HGB<11g/dl at onset and lymphocyte count<3×109/L, dysplastic megakarycytes≥10% (CD41 monoclonal antibogy immunohistochemical staining) and periodic acid schiff (PAS) staining (+) in bone marrow aspirate between RCC and NSAA (P=0. 034, 0. 025, 0. 032, 0. 029). Compared with NSAA, there was significant differences in the distribution of bone marrow cellularity down to 10% of the normal age matched value, dysplastic erythroblastic island, number of erythroblastic mitoses and micromegakaryocytes in bone marrow biopsy of RCC(P< 0. 0001, 0. 0001, 0. 0001, 0. 0001). Using Logistic regression, a new diagnostic scoring model including increased dysplastic erythroblastic island, having micromegakaryocytes in bone marrow biopsy, and lymphocyte count<3×109/L in peripheral blood was developed. By ROC curve analysis, score≥2 points was identified as a cut-off value with sensitivity of 70. 3% and specificity of 95%. Conclusion: RCC patients had similar clinical features with NSAA patients but had lower OS and higher possibillity of clonal evolution. This new diagnostic scoring model had significance to differentiate diagnose RCC and NSAA. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Investigating Patient Characteristics and Outcomes of Myelofibrosis Patients with Hyperleukocytosis

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 3041-3041
Author(s):  
Savan Shah ◽  
Chetasi Talati ◽  
Najla Al Ali ◽  
Eric Padron ◽  
David A Sallman ◽  
...  
Keyword(s):  
Overall Survival ◽  
Research Funding ◽  
Management Strategies ◽  
Myeloproliferative Neoplasm ◽  
Primary Myelofibrosis ◽  
Disease Diagnosis ◽  
Extramedullary Hematopoiesis ◽  
Myelogenous Leukemia ◽  
Similar Proportion ◽  
Increased Risk

Abstract Background: Primary myelofibrosis (PMF) is a myeloproliferative neoplasm that is marked by bone marrow fibrosis, cytopenias, extramedullary hematopoiesis, and a increased risk of acute leukemia. Leukocytosis is frequently seen and a white blood cell count (WBC) > 25,000/uL has been shown to convey an inferior prognosis. Rarely, patients develop severe leukocytosis (defined as WBC > 50,000/uL) either at presentation or during the course of their disease. Genomic characterization, management, and outcomes of these patients are not well defined. In this study, we aimed to further characterize PMF patients with severe leukocytosis and assess their clinical outcomes. Methods: We retrospectively reviewed PMF patients who presented to our institution between 2001 and 2018. We included patients who developed a WBC count > 50K at any time during their clinical course. Patients who developed acute myelogenous leukemia (AML) were censored at the time of disease diagnosis. Overall survival (OS) defined from first date that WBC documented as > 50K. Results: Among 493 PMF patients treated at our institution, 71 (14.4%) developed severe leukocytosis during the course of their disease. Ten (14%) had severe leukocytosis at the time of diagnosis and 30 (42%) developed it within 1 year of diagnosis and 40 (56%) developed it more than 1 year after diagnosis. Eight (11.3%) patients demonstrated peripheral blast percentage > 10% blasts at the time severe leukocytosis first documented. Compared to patients who did not develop severe leukocytosis, those with severe leukocytosis had an increased frequency of EZH2 (p < 0.001), RAS (p < 0.001), and KIT (p = 0.04) mutations. ASXL1 mutations were seen in a similar proportion of patients (p = 0.41). A similar proportion of patients were high risk by GIPSS (26% v 24%) prognostic model. From the time development of severe leukocytosis, the median overall survival (mOS) was 13.3 months. Median OS from diagnosis was significantly shorter for patients who developed severe leukocytosis (35.4 mo v 63.5 mo; p = 0.02) compared to those that did not. Eleven (15%) patients with severe leukocytosis developed AML compared to 38 (9%) patients who did not develop severe leukocytosis (p = 0.13). Median time to development of AML was 13.3 months from time of severe leukocytosis. At development of severe leukocytosis, 12 patients (17%) received hydrea (mOS 14.0 months) and 9 patients (13%) received ruxolitinib (mOS 21.2 months). The remaining cohort which either received no documented treatment, an alternative treatment or unknown treatment had a mOS of 5.3 months. This was not statistically significant. Ten patients (14%) ultimately underwent allogeneic stem cell transplantation after the development of hyperleukocytosis. Conclusion: Severe leukocytosis occurs rarely in primary myelofibrosis. RAS, KIT, and EZH2 mutations are enriched in these patients. Management strategies in these patients is varied and outcomes are poor. Further studies assessing the benefit of cytoreductive therapies in this population should be performed. Figure Figure. Disclosures Sallman: Celgene: Research Funding, Speakers Bureau. Sweet:BMS: Honoraria; Astellas: Consultancy; Jazz: Speakers Bureau; Novartis: Consultancy, Honoraria, Speakers Bureau; Jazz: Speakers Bureau; Phizer: Consultancy; BMS: Honoraria; Celgene: Honoraria, Speakers Bureau; Astellas: Consultancy; Agios: Consultancy; Phizer: Consultancy; Celgene: Honoraria, Speakers Bureau; Novartis: Consultancy, Honoraria, Speakers Bureau; Agios: Consultancy. List:Celgene: Research Funding. Komrokji:Novartis: Honoraria, Speakers Bureau; Celgene: Honoraria, Research Funding; Celgene: Honoraria, Research Funding; Novartis: Honoraria, Speakers Bureau; Novartis: Honoraria, Speakers Bureau; Novartis: Honoraria, Speakers Bureau. Kuykendall:Janssen: Consultancy; Celgene: Honoraria.

scholarly journals Plasma EDA Fibronectin in Primary Myelofibrosis Correlates with Bone Marrow Fibrosis and Predicts Splenomegaly

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 1688-1688
Author(s):  
Alessandro Malara ◽  
Cristian Gruppi ◽  
Margherita Massa ◽  
Vittorio Rosti ◽  
Giovanni Barosi ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Conflicts Of Interest ◽  
Philadelphia Chromosome ◽  
Myeloproliferative Neoplasm ◽  
Primary Myelofibrosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Costal Margin ◽  
Cell Transplant ◽  
Healthy Controls ◽  
Hematopoietic Stem

Introduction: Primary myelofibrosis (PMF) is a Philadelphia chromosome negative myeloproliferative neoplasm with adverse prognosis characterized by bone marrow (BM) fibrosis and extramedullary hematopoiesis. Fibronectin (FN) is an extracellular matrix glycoprotein that plays vital roles during tissue repair and regeneration. It exists in different forms. Plasma FN is synthesized by hepatocytes and secreted into the blood plasma, where circulates at a concentration of 300-600 μg/ml in a soluble, compact form. Differently, cellular FN is synthesized by several cell types, such as fibroblasts, endothelial cells, chondrocytes and myocytes. The alternative splicing of EDA and EDB and more complex splicing of the V domain, during transcription of FN1 gene, allows different isoforms of FN to be expressed in a tissue-dependent and temporally regulated manner. Very low levels (1.3-3 μg/ml) of FN containing EDA and/or EDB are present in plasma. Although its function is not well understood, EDA containing FN (EDA-FN) is known to agonize Toll like receptor 4 (TLR4), resulting in NF-κβ-dependent cytokine release; to induce myofibroblast differentiation during wound healing; and to increase agonist-induced platelet aggregation and thrombus formation in vivo. We previously showed that EDA-FN levels are increased in plasma and BM biopsies of PMF patients. Mechanistically, BM EDA-FN sustains megakaryocyte proliferation through TLR4 binding and confer a pro-inflammatory phenotype to cell niches promoting fibrosis progression in Romiplostim-treated mice. In this work we measured the plasma levels of EDA-FN in 104 well characterized patients with PMF to determine whether elevated levels of EDA-FN predict the occurrence of disease-related events. Methods: Plasma circulating EDA FN was measured with an enzyme linked immunosorbent assay developed at the University of Pavia, by our group. We obtained plasma EDA-FN concentration values and health care data of persons with PMF from the data-base of the Centre for the Study of Myelofibrosis at the IRCCS Policlinico S. Matteo Foundation in Pavia. We sequentially excluded persons treated with disease-modifying drugs at any time before or on the date of base-cohort entry, and those who had been splenectomized or had received a stem cell transplant. We also excluded persons with acute inflammatory diseases, autoimmune diseases, other neoplasms, and severe liver or renal dysfunction. For this study we selected everyone giving written informed consent and the study was approved by the local Ethic Committee. Immunofluorescence was performed on spleen sections from PMF patients who underwent splenectomy either because of anemia or symptomatic splenomegaly, or both; and healthy controls that were splenectomized following traumatic lesion of the spleen. Data were analyzed using STATISTICA software. Results: A homozygous JAK2V617F genotype was the major determinant of elevated plasma EDA-FN. Elevated EDA-FN levels were associated with anemia, increased levels of high-sensitivity C-reactive protein, BM fibrosis and splanchnic vein thrombosis at diagnosis. We interpreted these associations as reflecting the role EDA-FN plays in tissue remodeling, inflammation and vascular injury. Interestingly, EDA-FN levels resulted also associated with spleen size, and elevated levels of EDA-FN at diagnosis predicted large splenomegaly (more than 10 cm from the left costal margin) outcome. The evidence that plasma EDA-FN levels were not associated with the CD34+ hematopoietic stem cells mobilization, drove us to hypothesize that EDA-FN could reflect spleen endothelial cell activation and/or neoangiogenesis. Immunofluorescence analysis of spleen specimens from PMF patients and healthy controls revealed that high levels of EDA-FN were present in pathological spleens in strong association with endothelial neoangiogenesis. Conclusions: Quantification of EDA-FN level in PMF strongly correlates with BM fibrosis and may be the first marker of an altered spleen microvasculature that contributes to splenomegaly. Understanding the role of this FN isoform in PMF would be useful for testing new mechanisms of disease progression and new hypotheses about the treatment of splenomegaly in PMF. Disclosures No relevant conflicts of interest to declare.

scholarly journals Endogenous TGF-beta1 Mediated Osteogenic Impairment of Medullar Mesenchymal Stromal Cells in Primary Myelofibrosis

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 1873-1873
Author(s):  
Christophe Martinaud ◽  
Christophe Desterke ◽  
Johanna Konopacki ◽  
Lisa Pieri ◽  
Rachel Golub ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Conflicts Of Interest ◽  
Myeloproliferative Neoplasm ◽  
Hematopoietic Cells ◽  
Primary Myelofibrosis ◽  
Osteogenic Potential ◽  
Healthy Donors

Abstract Primary myelofibrosis (PMF) is myeloproliferative neoplasm characterized by clonal myeloproliferation, dysmegakaryopoiesis, extramedullary hematopoiesis associated with myelofibrosis and altered stroma in bone marrow and spleen. Mesenchymal stromal cells (MSCs) are reported to play a pivotal role in fibrosis and stromal changes are considered as a reactive counterpart of the cytokine production by clonal hematopoietic cells. The present study shows that MSCs from patients demonstrate functional abnormalities that are unexpectedly maintained ex-vivo, in culture. Material and Methods: we studied MSCs and bone marrow sections from PMF patients (n=12) as compared to healthy donors (HDs) (n=6). We tested their proliferation, immunophenotype, hematopoiesis supporting capacities, differentiation abilities, in-vivo osteogenic assays, and performed secretome and transcriptome analysis. Results: We found that PMF-MSCs exhibit similar proliferative capacity and long-term hematopoiesis supporting abilities as compare to healthy donors. They overproduce interleukin 6, VEGF, RANTES, PDGF, BMP-2 and surprisingly TGF-beta1. MSCs from fibrotic PMF patients express high levels of glycosaminoglycans. Adipocytes and chondrocytes differentiation abilities were not different as compared to HDs but PMF-MSCs exhibit an increased in vitro potential. Implementation on scaffold in nude mice confirmed, in vivo, this increased osteogenic potential. We then looked into gene expression and discovered that PMF-MSCs show an original transcriptome signature related to osteogenic lineage and TGF-beta1. Indeed, osteogenic genes such as Runx2, Dlx5, Twist1, Noggin, Sclerostin, GDF5 and Serpine1 are deregulated and suggest a potential osteoprogenitor priming of PMF-MSCs. These molecular results also advocated for a TGF-beta1 impregnation that prompted us to study its impact on PMF-MSCs osteogenic differentiation. First, we then showed that Smad2 is intrinsically over-activated in PMF-MSC and that stimulation by TGF-beta1 is associated with an increase phospho-Smad2 level and an enhancement of bone master gene regulator Runx2 expression. Then, we inhibited TGF-beta1 pathway by by SB-431542 and evidenced a specific behavior of osteogenic MSCs differentiation in patients, suggesting involvement of TGF-beta1 in osteogenic impairment. Conclusion: Altogether, our results identify a signature of PMF-MSCs and suggest that they participate in PMF osteogenic dysregulation independently from in vivo local stimulation by clonal hematopoietic cells Disclosures No relevant conflicts of interest to declare.

scholarly journals Macrophage frequency in the bone marrow correlates with morphologic subtype of myeloproliferative neoplasm

2020 ◽  
Vol 100 (1) ◽  
pp. 97-104
Author(s):  
David C. A. Molitor ◽  
Peter Boor ◽  
Andreas Buness ◽  
Rebekka K. Schneider ◽  
Lino L. Teichmann ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Myeloid Leukemia ◽  
Clinical Course ◽  
Myeloproliferative Neoplasms ◽  
Philadelphia Chromosome ◽  
Myeloproliferative Neoplasm ◽  
Primary Myelofibrosis ◽  
Neoplastic Processes ◽  
And Control

AbstractBone marrow (BM) fibrosis in myeloproliferative neoplasms (MPNs) is associated with a poor prognosis. The development of myelofibrosis and differentiation of mesenchymal stromal cells to profibrotic myofibroblasts depends on macrophages. Here, we compared macrophage frequencies in BM biopsies of MPN patients and controls (patients with non-neoplastic processes), including primary myelofibrosis (PMF, n = 18), essential thrombocythemia (ET, n = 14), polycythemia vera (PV, n = 12), and Philadelphia chromosome–positive chronic myeloid leukemia (CML, n = 9). In PMF, CD68-positive macrophages were greatly increased compared to CML (p = 0.017) and control BM (p < 0.001). Similar findings were observed by CD163 staining (PMF vs. CML: p = 0.017; PMF vs. control: p < 0.001). Moreover, CD68-positive macrophages were increased in PV compared with ET (p = 0.009) and reactive cases (p < 0.001). PMF had higher frequencies of macrophages than PV (CD68: p < 0.001; CD163: p < 0.001) and ET (CD68: p < 0.001; CD163: p < 0.001). CD163 and CD68 were often co-expressed in macrophages with stellate morphology in Philadelphia chromosome–negative MPN, resulting in a sponge-like reticular network that may be a key regulator of unbalanced hematopoiesis in the BM space and may explain differences in cellularity and clinical course.

scholarly journals Prefibrotic Primary Myelofibrosis As an Entity Distinct from Other Philadelphia-Chromosome Negative Myeloproliferative Neoplasms - Data from the Austrian Reclassification Project

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 4252-4252 ◽  
Author(s):  
Georg Jeryczynski ◽  
Bettina Gisslinger ◽  
Martin Schalling ◽  
Maria Theresa Krauth ◽  
Leonhard Muellauer ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Myeloproliferative Neoplasms ◽  
Survival Rates ◽  
Philadelphia Chromosome ◽  
Myeloproliferative Neoplasm ◽  
Primary Myelofibrosis ◽  
Distinct Entity ◽  
Bone Marrow Biopsies ◽  
Cumulative Rate ◽  
Night Sweats

Abstract Introduction: The distinction between essentialthrombocythemia(ET) and early, prefibrotic primary myelofibrosis (prePMF) by strictly applying the criteria of the WHO classification results in a newly defined subgroup of myeloproliferative neoplasm (MPN) in which the frequency and severity of clinical features are currently poorly characterized. Since prePMF was usually summarized under the subgroup of ET in previous classifications, little is known about the frequency of prePMFamong MPN. The importance of accurate diagnosis, however, is underlined by various publications that could demonstrate a significant impact on management and outcome of these patients. Aims: The aim of this study was to describe the clinical characteristics and symptoms at diagnosis of prePMF as a distinct entity and to evaluate the course of the disease in regard to survival, transformation into overt myelofibrosis and acute leukemia. Methods: All patients with representative bone marrow biopsies at presentation and complete clinical data at time of diagnosis and follow up were included in this study. All patients included were recruited from the MPN cohort of the Medical University of Vienna. Results: In total, 807 MPN patients diagnosed according to the WHO 2008 criteria were analyzed. The relative frequency of prePMF in our cohort was 17.6% (n=142) as opposed to 27.4% (221) in ET (Fig. 1). The median age for prePMF patients was 63.4 years (range 26.9-88.1) and 55.1% were female. The majority showed an elevated platelet count (median 770 G/l, range 78-2869), hemoglobin levels were on the lower end of the normal range in both genders (median 13.9, range 8.1-18.4 and 13.2, range 7.9-16.2 in men and women respectively). Leukocyte counts in the upper range of normal were common (median 9.87, range 4.0-46.0). The lactate dehydrogenase levels (LDH) were markedly elevated (median 303, range 153-729 with an institutional cut-off of 250 U/L). Therewasalso a considerate proportion of patients with splenomegaly (39.8%). Further, 22.6% of patients reported constitutional symptoms such as night sweats and weight loss. Fiber grade of 1 of a three-graded score in the bone marrow biopsy was reported in 26.2% of cases. 27.0% of patients presented withleftshiftingwith a few peripheral blasts. JAK2 positivity was found in 57.1%, CALR in 32.7 and MPL in 3.3% of cases. Only 5.9% were triple negative. 5-, 10- and 20-year survival rates were 87.6%, 67.0% and 28.8% respectively (38.2% of patients followed to death). Cumulative rate of progression into overt fibrosis was 34.2% after 10 and 58.8% after 20 years (Fig. 2). Discussion: Our data highlight important features of prePMF. Firstly, compared to ET, it has a later onset. More importantly, thrombocytosis is not a feature limited to ET, but is also frequently seen in prePMF and is therefore not suitable to accurately characterize MPN at time of diagnosis. However, leukocytosis and elevated LDH levels are features uncommon for ET as is fiber grade > 0 that is almost never seen in the WHO-defined ET, but relatively common in prePMF. Splenomegaly and presence of a few blasts are generally signs of beginningextramedullaryhematopoiesis are therefore more commonly associated with early stages primary myelofibrosis. Lastly, overall survival and fibrosis-free survival is substantially impaired in prePMF in comparison to data previously published for ET and similar to polycythemiavera. In conclusion, prePMF is a distinct entity, that while sharing some features of ET, most notably thrombocytosis, shows several striking features that should be regarded when investigating newly diagnosed MPN patients. Figure Figure. Figure Figure. Disclosures Gisslinger: Baxalta: Consultancy, Honoraria; Novartis: Consultancy, Honoraria; AOP Orphan: Consultancy, Honoraria.

Point Mutations In Myelodysplastic Syndromes Are Associated with Clinical Features and Are Independent Predictors of Overall Survival

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 300-300
Author(s):  
Rafael Bejar ◽  
Kristen Stevenson ◽  
Omar Abdel-Wahab ◽  
McConkey Marie ◽  
Katherine Lin ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Overall Survival ◽  
Clinical Practice ◽  
Clinical Features ◽  
Point Mutations ◽  
Scoring Systems ◽  
Bone Marrow Blast ◽  
Oncogenic Mutations ◽  
Normal Cytogenetics ◽  
Prognostic Scoring Systems

Abstract Abstract 300 Myelodysplastic syndromes (MDS) are clonal hematopoietic disorders with highly variable clinical features. Much of this heterogeneity is likely driven by the diverse set of genetic lesions associated with MDS. Prognostic scoring systems help stratify patients into risk groups based on clinical measures, bone marrow blast counts, and common cytogenetic abnormalities. However, the presence or absence of point mutations is not considered by the prognostic scoring systems used in clinical practice. Since more than half of MDS cases have a normal karyotype, genetic abnormalities do not contribute to prognostic risk in the majority of patients. In order to better understand the frequency, overlap, and clinical impact of point mutations in MDS, we examined samples from a cohort of 438 patients for mutations in cancers genes. First we screened 191 samples for the presence of 1233 known oncogenic mutations in over 130 cancer-related genes using a high-throughput, mass spectroscopic genotyping platform (OncoMap). Somatic mutations were validated in 7 genes. Known oncogenic mutations in these genes were then sought in an expanded cohort of 438 MDS patient samples. Additional mutated samples were identified for 6 of these 7 genes, including NRAS, KRAS, BRAF, and JAK2. Our second approach utilized next-generation 454-pyrosequencing of several tumor suppressor genes and MDS-related genes not covered with OncoMap, including TET2, RUNX1, TP53, CBL, NPM1, PTEN and CDNK2A. This was complemented by Sanger sequencing of additional genes including IDH1, IDH2, ASXL1, and KDM6A. In aggregate, 50.9% of samples were found to carry at least one mutation, including 50.8% of samples with normal cytogenetics. The most frequently mutated genes were TET2 (18%), ASXL1 (14%), RUNX1 (8%), and TP53 (7%). Recurring mutations in NRAS, JAK2, IDH1, IDH2, CBL, NPM1, KDM6A, and KRAS were identified at lower frequency. TP53 mutations were largely exclusive of all other mutations except for those in TET2 and TP53 mutations were highly associated with complex cytogenetics and abnormalities of chromosome 17. TET2 mutations were overrepresented in cases of normal cytogenetics but not predictive of survival, even after stratification by mutant allele burden. Mutations of RUNX1, NRAS, and TP53 were each associated with a lower platelet count (p<0.001 for each comparison). Mutations in these genes and CBL were also associated with a higher bone marrow blast count (p≤0.01 for each comparison). Several genes, including NRAS, RUNX1, TP53, CBL, IDH2, and ASXL1 were associated with decreased overall survival (p=0.01, p<0.001, p<0.001, p=0.02, p=0.03, p=0.01, respectively). In a multivariable model including age, sex, and International Prognostic Scoring System risk group, RUNX1 (hazard ratio [HR], 1.61; 95% confidence interval [CI], 1.10–2.35), TP53 (HR, 2.34; 95% CI, 1.50–3.64), and ASXL1 (HR, 1.42; 95%, CI1.05-1.92) were independent predictors of decreased overall survival. In summary, we performed a broad survey for gene mutations associated with myeloid neoplasms or other cancers. In our cohort of 438 clinically annotated MDS patient samples, we identified point mutations in over 15 genes with more than 50% of samples harboring at least one mutation. Mutations in several genes were associated with clinical features of MDS including thrombocytopenia and elevated blast counts. Mutations of RUNX1, TP53, and ASXL1 (present in 26.3% of samples) are independent predictors of decreased survival, demonstrating that incorporation of point mutations adds information to the risk stratification systems used in clinical practice. Disclosures: No relevant conflicts of interest to declare.

Molecular Genetic Analysis of Myelodysplastic Syndromes (MDS) Patients with Ring Sideroblasts (RS); Independent Confirmation of Association of SF3B1 Mutations with Better Prognosis

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 3237-3237
Author(s):  
Abdullah Mahmood Ali ◽  
Nicholas Iverson ◽  
Alexander Penson ◽  
Allison Scott ◽  
Tabish Riaz ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Overall Survival ◽  
Genetic Analysis ◽  
Myelodysplastic Syndromes ◽  
Molecular Genetic ◽  
Jak2 V617f ◽  
Molecular Genetic Analysis ◽  
Kaplan Meier ◽  
Ring Sideroblasts ◽  
Calr Mutations

Abstract Background: Ring sideroblasts (RS) are erythroblasts with iron-loaded mitochondria, appears as blue granules when stained with Prussian blue, and are characteristic of myelodysplastic syndromes (MDS) subgroups refractory anemia with ring sideroblasts (RARS), refractory cytopenia with multilineage dysplasia and ring sideroblasts (RCMD-RS), and RARS with thrombocytosis (RARS-T) but are also infrequently found in other MDS groups. The etiology of RS remains unclear. Molecular genetic analysis of MDS patients with RS (MDS-RS) using exome and targeted sequencing identified mutations in several genes including SF3B1. While the RS phenotype was shown to be strongly associated with SF3B1 mutations, the association of SF3B1 mutations with better prognosis remained controversial. Also, there remained several patients who do not show any mutations in SF3B1 suggesting there are additional genes associated with RS phenotype. In addition to SF3B1 mutations, RARS-T patients show mutations in JAK2 (V617F and Exon 12), MPL and CALR genes, frequently mutated in Essential Thrombocytopenia (ET). But there remained a high number of patients who do not show any mutation in JAK2, MPL and CALR genes suggesting additional genes are associated with thrombocytosis. In order to gain further insight into the molecular genetics of MDS-RS patients and to elucidate their clinical significance we screened a large cohort of patients with RS (RARS/RCMD-RS) and a subgroup with thrombocytosis (RARS-T). Methods: This study is approved by the Institutional Review Board of Columbia University and informed consent was obtained from all the individuals participated in the study. Bone marrow mono nuclear cells were isolated from bone marrow aspirate and DNA was extracted using DNeasy Blood and Tissue kit. To screen for most frequent mutations in SF3B1 (exon 14 and 15), MPL (exon 10), JAK2 (exon 12), and CALR (exon 9) genes, primers were designed to amplify the exons and the exon-intron junctions using PCR and the amplified and purified PCR products were sequenced using Sanger sequencing on both strands. To screen for mutations V617F in JAK2, primers were designed to carry out allele specific PCR. Demographic and clinical data is collected from patient’s charts/reports. Statistical analysis including survival curve (Kaplan–Meier method) was performed using GraphPad Prism Software. Results: A total of 209 patients with RS phenotype were screened for SF3B1 mutations. Mutations in SF3B1 gene were detected in 62% (130/209) of RS patients, including 85% (23/27) of RARS-T and 59% (107/182) of RARS/RCMD-RS patients. Among all the SF3B1 mutations, K700E was the most frequent mutation (60%) followed by mutations at H662. Clinical significance of SF3B1 mutations on overall survival, using Kaplan-Meier method, showed SF3B1 mutations were associated with better prognosis (Fig 1). The median survival of patients with SF3B1 mutation is 110 months compared to those without mutations, 70 months (p value < 0.034). Studies to screen SF3B1 negative patients for additional mutations are being carried out using exome sequencing to identify genes associated with RS phenotype. In addition to SF3B1, RARS-T patients were also screened for JAK2 V617F and exon 12 mutations. JAK2 V617F mutations were detected in 11% of RARS-T patients and no mutations were found in JAK2 exon 12. Patients negative for JAK2 were screened for mutations in MPL and CALR genes. No mutations were found in MPL but 8% (2/23) of those negative for JAK2 were found to have CALR mutations. Both CALR mutations were frameshift mutations that alter the C-terminus thus abolishing the KDEL sequence required for CALR function. There are still 80% RARS-T patients in our cohort who do not show any mutations in JAK2, MPL, CALR genes. Conclusions: The association of SF3B1 mutation with prognosis is controversial with some studies suggesting a better prognosis while other reports no effect. Our study, using a large cohort of well-characterized RS patients provides an Independent verification of the observation that SF3B1 mutations are associated with better overall survival. There remains a large group of patients (40% in RARS/RCMD-RS groups) without SF3B1 mutation but with a RS phenotype suggesting yet other gene(s) is associated with RS phenotype. Similarly, there remain patients who do not show any mutations in JAK2, MPL and CALR suggesting other genes are involved in the etiology of thrombocytopenia. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

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