Diagnosis and treatment of systemic mastocytosis in Brazil: Recommendations of a multidisciplinary expert panel

ABSTRACT Introduction: Systemic Mastocytosis comprises a group of neoplastic diseases characterized by clonal expansion and infiltration of mast cells into several organs. The diagnosis and treatment of this disease may be challenging for non-specialists. Objective: Make suggestions or recommendations in Systemic Mastocytosis based in a panel of Brazilian specialists. Method and results: An online expert panel with 18 multidisciplinary specialists was convened to propose recommendations on the diagnosis and treatment of Systemic Mastocytosis in Brazil. Recommendations were based on discussions of topics and multiple-choice questions and were graded using the Oxford Centre for Evidence-Based Medicine 2011 Levels of Evidence Chart. Conclusion: Twenty-two recommendations or suggestions were proposed based on a literature review and graded according to the findings.

Philadelphia-negative myeloproliferative neoplasms as disorders marked by cytokine modulation

ABSTRACT Background: Cytokines are key immune mediators in physiological and disease processes, whose increased levels have been associated with the physiopathology of hematopoietic malignancies, such as myeloproliferative neoplasms. Methods: This study examined the plasma cytokine profiles of patients with essential thrombocythemia, primary myelofibrosis, polycythemia vera and of healthy subjects, and analyzed correlations with JAK2 V617F status and clinical-hematological parameters. Results: The proinflammatory cytokine levels were increased in myeloproliferative neoplasm patients, and the presence of the JAK2 V617F mutation was associated with high IP-10 levels in primary myelofibrosis patients. Conclusions: Essential thrombocythemia, primary myelofibrosis, and polycythemia vera patients exhibited different patterns of cytokine production, as revealed by cytokine network correlations. Together, these findings suggest that augmented cytokine levels are associated with the physiopathology of myeloproliferative neoplasms.

Insulin Substrate Receptor (IRS) proteins in normal and malignant hematopoiesis

The insulin receptor substrate (IRS) proteins are a family of cytoplasmic proteins that integrate and coordinate the transmission of signals from the extracellular to the intracellular environment via transmembrane receptors, thus regulating cell growth, metabolism, survival and proliferation. The PI3K/AKT/mTOR and MAPK signaling pathways are the best-characterized downstream signaling pathways activated by IRS signaling (canonical pathways). However, novel signaling axes involving IRS proteins (noncanonical pathways) have recently been identified in solid tumor and hematologic neoplasm models. Insulin receptor substrate-1 (IRS1) and insulin receptor substrate-2 (IRS2) are the best-characterized IRS proteins in hematologic-related processes. IRS2 binds to important cellular receptors involved in normal hematopoiesis (EPOR, MPL and IGF1R). Moreover, the identification of IRS1/ABL1 and IRS2/JAK2V617F interactions and their functional consequences has opened a new frontier for investigating the roles of the IRS protein family in malignant hematopoiesis. Insulin receptor substrate-4 (IRS4) is absent in normal hematopoietic tissues but may be expressed under abnormal conditions. Moreover, insulin receptor substrate-5 (DOK4) and insulin receptor substrate-6 (DOK5) are linked to lymphocyte regulation. An improved understanding of the signaling pathways mediated by IRS proteins in hematopoiesis-related processes, along with the increased development of agonists and antagonists of these signaling axes, may generate new therapeutic approaches for hematological diseases. The scope of this review is to recapitulate and review the evidence for the functions of IRS proteins in normal and malignant hematopoiesis.

TET2 is upregulated during erythroid differentiation of CD34+ cells from healthy donors and myelodysplastic syndrome patients

Background: Tet methylcytosine dioxygenase 2 (TET2) is frequently mutated and/or downregulated in myeloid neoplasm, including myelodysplastic syndromes. Despite the extensive studies, the specific contribution of TET2 in disease phenotype of myeloid neoplasms is not fully elucidated. Recent findings have grown attention on the role of TET2 in normal and malignant erythropoiesis. Methods: In the present study, we investigated TET2 mRNA levels by quantitative PCR during erythropoietin-induced erythroid differentiation CD34+ cells from healthy donor and myelodysplastic syndrome patients. Statistical analyses were performed using the ANOVA and Bonferroni post hoc test and a p-value <0.05 was considered statically significant. Results: TET2 expression is upregulated during erythroid differentiation of CD34+ cells from healthy donor and myelodysplastic syndrome patients. Conclusions: Our findings corroborate that TET2 is involved in the erythrocyte differentiation (AU)

IL32 expression in peripheral blood CD3+ cells from myelodysplastic syndromes patients

Background: Myelodysplastic syndromes (MDS) are a heterogeneous group of disorders characterized by ineffective hematopoiesis and risk of leukemia transformation. There is evidence to suggest the participation of immune system deregulation in MDS pathogenesis. Interleukin-32 (IL-32) is a newly described multifunctional cytokine reported as an important mediator in autoimmune and inflammatory disorders. In the present study, we reported the expression of IL32 and IL32 transcript variants (α, ß, γ and δ) in peripheral blood CD3+ cells from healthy controls and MDS patients. Methods: CD3+ cells were isolated by immunomagnetic cell sorting from thirty-nine untreated MDS patients and twenty-nine healthy donors. Gene expression was evaluated by quantitative PCR. For statistical analysis, Mann­Whitney test, Kruskal-Wallis test with Dunns post test and Log-rank (Mantel-Cox) were used, as appropriate. A p value <0.05 was considered statistically significant. Results: IL32 expression and IL32 transcript variants IL32α, IL32ß, IL32γ, and IL32δ, were similar in peripheral blood CD3+ cells from healthy donors and MDS patients. Increased IL-32α expression was an independent predictor for MDS disease progression by univariate and multivariate analysis. Conclusions: We observed that IL32 expression is not differently expressed in CD3+ cells from MDS patients; nevertheless IL32α has a potential role in disease progression (AU)

SIVA, a target of p53, is downregulated in myelodysplastic syndromes

Background: SIVA is a transcriptional target of p53 that plays a potential role in the development and progression of cancer. In this study, we analyzed SIVA1 and SIVA2 expression, and its association with clinical features and TP53 and MDM2 expression in bone marrow cells from healthy donors and myelodysplastic syndrome (MDS) patients. Methods: Fifty-five untreated patients with MDS and 22 healthy donors were included. Gene expression was evaluated by quantitative PCR. For statistical analysis, Mann­Whitney test, Spearman correlation analysis and Log-rank (Mantel-Cox) were used, as appropriate. A p value <0.05 was considered statistically significant. Results: SIVA1 and SIVA2 transcripts were significantly decreased in bone marrow samples from MDS patients compared to healthy donors, and positively correlated with MDM2 and TP53 expression in MDS patients (all p < 0.05). MDM2 expression was also downregulated in bone marrow samples from MDS patients compared to healthy donors (p < 0.05). However, SIVA1, SIVA2, MDM2 and TP53 expressions did not impact on MDS outcomes. Conclusions: SIVA1 and SIVA2 transcripts are downregulated in bone marrow samples from MDS patients (AU)

Myelodysplastic syndrome with synchronous gastric cancer: when the symptoms suggest something else

Although myelodysplastic syndromes have a clear definition in theory, the morphologic dysplasia associated with ineffective hematopoiesis may be subtle and difficult to recognize and can commonly be mimicked by systemic conditions, such as infections, autoimmune disorders, nutritional deficiencies, toxic factors and non-hematological malignancies. However, myelodysplastic syndromes may truly coexist with other systemic diseases, which can be masked when the patient's symptoms are attributed exclusively to myelodysplastic syndromes without further investigation. To better illustrate this, we herein describe two cases associated with synchronous gastric cancers...

PTK2 and PTPN11 expression in myelodysplastic syndromes

OBJECTIVE: The aim of this study was to evaluate the expression of protein tyrosine kinase 2 and protein tyrosine phosphatase non-receptor type 11, which respectively encode focal adhesion kinase protein and src homology 2 domain-containing protein-tyrosine phosphatase 2, in hematopoietic cells from patients with myelodysplastic syndromes. METHODS: Protein tyrosine kinase 2 and tyrosine phosphatase non-receptor type 11 expressions were analyzed by quantitative polymerase chain reaction in bone marrow cells from patients with myelodysplastic syndromes and healthy donors. RESULTS: Protein tyrosine kinase 2 and tyrosine phosphatase non-receptor type 11 expressions did not significantly differ between normal cells and myelodysplastic cells. CONCLUSIONS: Our data suggest that despite the relevance of focal adhesion kinase and src homology 2 domain-containing protein-tyrosine phosphatase 2 in hematopoietic disorders, their mRNA expression do not significantly differ between total bone marrow cells from patients with myelodysplastic syndromes and healthy donors. .

Screening for hotspot mutations in PI3K, JAK2, FLT3 and NPM1 in patients with myelodysplastic syndromes

INTRODUCTION: Myelodysplastic syndromes encompass a heterogeneous group of clonal hematopoietic stem cell disorders characterized by ineffective hematopoiesis, refractory cytopenia and a tendency to progress toward acute myeloid leukemia. The accumulation of genetic alterations is closely associated with the progression of myelodysplastic syndromes toward acute myeloid leukemia. OBJECTIVE: To investigate the presence of mutations in the points most frequent for mutations (hotspot mutations) in phosphatidylinositol-3-kinase (PI3K), Janus kinase 2 (JAK2), FMS-like tyrosine kinase 3 (FLT3) and nucleophosmin (NPM1), which are involved in leukemia and other cancers, in a population of Brazilian MDS patients. METHODS: Fifty-one myelodysplastic syndromes patients were included in the study. According to French-American-British classification, the patients were distributed as follows: 31 with refractory anemia, 8 with refractory anemia with ringed sideroblasts, 7 with refractory anemia with excess blasts, 3 with refractory anemia with excess blasts in transformation and 2 with chronic myelomonocytic leukemia. Bone marrow samples were obtained and screened for the presence of hotspot mutations using analysis based on amplification with the polymerase chain reaction, sequencing, fragment size polymorphisms or restriction enzyme digestion. All patients were screened for mutations at the time of diagnosis, and 5 patients were also screened at the time of disease progression. RESULTS: These results show that hotspot mutations in the PI3K, JAK2, FLT3 and NPM1 genes are not common in MDS patients; nevertheless, JAK2 mutations may be present in myelodysplasia during disease progression. CONCLUSIONS: These results show that hotspot mutations in the PI3K, JAK2, FLT3 and NPM1 genes are not common in MDS patients; nevertheless, JAK2 mutations may be present in myelodysplasia during disease progression.

Low bone mass density is associated with hemolysis in brazilian patients with sickle cell disease

OBJECTIVES: To determine whether kidney disease and hemolysis are associated with bone mass density in a population of adult Brazilian patients with sickle cell disease. INTRODUCTION: Bone involvement is a frequent clinical manifestation of sickle cell disease, and it has multiple causes; however, there are few consistent clinical associations between bone involvement and sickle cell disease. METHODS: Patients over 20 years of age with sickle cell disease who were regularly followed at the Hematology and Hemotherapy Center of Campinas, Brazil, were sorted into three groups, including those with normal bone mass density, those with osteopenia, and those with osteoporosis, according to the World Health Organization criteria. The clinical data of the patients were compared using statistical analyses. RESULTS: In total, 65 patients were included in this study: 12 (18.5 percent) with normal bone mass density, 37 (57 percent) with osteopenia and 16 (24.5 percent) with osteoporosis. Overall, 53 patients (81.5 percent) had bone mass densities below normal standards. Osteopenia and osteoporosis patients had increased lactate dehydrogenase levels and reticulocyte counts compared to patients with normal bone mass density (p<0.05). Osteoporosis patients also had decreased hemoglobin levels (p<0.05). Hemolysis was significantly increased in patients with osteoporosis compared with patients with osteopenia, as indicated by increased lactate dehydrogenase levels and reticulocyte counts as well as decreased hemoglobin levels. Osteoporosis patients were older, with lower glomerular filtration rates than patients with osteopenia. There was no significant difference between the groups with regard to gender, body mass index, serum creatinine levels, estimated creatinine clearance, or microalbuminuria. CONCLUSION: A high prevalence of reduced bone mass density that was associated with hemolysis was found in this population, as indicated by the high lactate dehydrogenase levels, increased reticulocyte counts and low hemoglobin levels.

Deficiência de ferro no paciente submetido à ressecção gástrica ou intestinal: prevalência, causas, repercussões clínicas, abordagem diagnóstica e prevenção: [revisão] / Iron deficiency anemia in patients submitted to gastric or small intestine resection: prevalence, cause, clinical outcome, diagnostic workup and prevention: [review]

Anemia ferropriva é frequente em pacientes submetidos à ressecção gástrica, para o tratamento de úlcera péptica ou câncer gástrico, e à cirurgia bariátrica, para o tratamento de obesidade. As cirurgias bariátricas podem ter como consequência a restrição do estômago, sem necessariamente ressecção gástrica, ou ressecção gástrica associada ou não a ressecção intestinal. As causas da deficiência de ferro nestes pacientes são multifatoriais e incluem: (1) baixa ingestão de ferro oral por intolerância a alimentos ricos em ferro; (2) redução da acidez gástrica pela perda de células parietais, dificultando a conversão do ferro férrico para a forma ferrosa; (3) exclusão do duodeno com consequente exclusão do principal sítio de absorção do ferro e restrição da ação das enzimas pancreáticas na liberação do ferro ligado ao heme. A investigação e a reposição de ferro de forma eficaz e precoce pode evitar essa complicação. Pacientes refratários ao ferro oral podem necessitar de reposição de ferro parenteral e todos os pacientes requerem acompanhamento periódico com hemograma e estoques de ferro por toda a vida.

Iron deficiency anemia is associated with gastrectomy used in the treatment of peptic ulcers and gastric cancer, and also in bariatric surgery for the treatment of obesity. Bariatric surgery involves restrictive and bypass procedures, gastrectomy and small intestine resection. There are several reasons why iron deficiency occurs in these patients including: (1) diminished iron intake due to meat intolerance (2) diminished gastric acid secretion due to loss of parietal cells which impairs the solubilization of ferric iron to form ferrous iron and (3) exclusion of the duodenum with consequent elimination of the main site of iron absorption and restriction in the action of pancreatic enzymes to release iron bound to the heme. Early clinical evaluation and use of iron supplements may be effective to prevent iron deficiency anemia in this population. Patients who remain refractory to oral supplementation may require parenteral iron administration. All patients require periodical and lifelong follow-up of hematological and iron parameters.

Complicações hepáticas na doença falciforme / Hepatic complications in sickle cell disease

Doentes falciformes podem apresentar alterações hepáticas agudas ou crônicas. As agudas são caracterizadas por dor no quadrante superior direito e icterícia. O diagnóstico diferencial inclui crise aguda de falcização hepática, seqüestro hepático, colestase intra-hepática, colelitíase, coledocolitíase, colecistite e hepatite viral aguda. Estas complicações devem ser diagnosticadas precocemente, através de história clínica, testes de função hepática e exames radiológicos, e o tratamento deve ser prontamente iniciado. Transfusão sangüínea é essencial para o tratamento das manifestações agudas causadas pelo processo de vaso-oclusão, como seqüestro hepático e colestase intra-hepática. As alterações hepáticas crônicas são freqüentemente causadas pela hemólise crônica e múltiplas transfusões. Para prevenção, diagnóstico precoce e orientação terapêutica da alteração hepática crônica, os doentes falciformes devem ser submetidos a exames de rotina: testes de função hepática, sorologia para hepatite B e C, dosagem sérica de ferritina e ultra-sonografia de abdômen. A biópsia hepática deve ser realizada em pacientes com hepatite viral e em pacientes com alterações hepáticas crônicas acentuadas e persistentes, afora das manifestações agudas.

Patients with sickle cell disease may present acute or chronic hepatopathy. The acute syndrome is characterized by right upper quadrant abdominal pain and jaundice. The differential diagnoses include acute sickle hepatic crises, hepatic sequestration, sickle cell intrahepatic cholestasis, cholecystitis, choledocholithiasis and acute viral hepatitis. These alterations can be differentiated by a careful history, liver function tests and hepatobiliary imaging studies. The specific treatment must be promptly initiated. Red blood cell transfusion is essential for the treatment of the clinical syndromes caused by the sickling process such as hepatic sequestration and sickle cell intrahepatic cholestasis. Chronic liver disease is frequently caused by chronic hemolysis and multiple transfusions. In an attempt to prevent, early diagnosis and treatment of chronic liver disease, sickle cell disease patients must be routinely submitted to liver function tests, serologic tests for hepatitis B and C, serum ferritin levels and abdominal ultrasound. Liver biopsy may be indicated in patients with virus hepatitis and in patients with persistent and accentuated alterations in liver function tests, out of acute sickle cell hepatic crises.

Indicações de transplante de células-tronco hematopoéticas para pacientes com diagnóstico de síndromes mielodisplásicas / Indications for hematopoietic stem cell therapy for patients with diagnosis of myelodysplastic syndromes

O transplante de células-tronco hematopoéticas (TCTH) é uma opção de cura para pacientes com diagnóstico de síndromes mielodisplásicas (SMD). Porém, diante da alta morbidade e mortalidade relacionada ao procedimento, as indicações e o momento de realizar o TCTH para pacientes com SMD permanecem controversos. Baseado nas evidências disponíveis até o momento, para pacientes com IPSS de baixo risco e intermediário 1 com doador aparentado HLA-idêntico, o TCTH deveria ser adiado por algum período, mas realizado antes do desenvolvimento de LMA, por exemplo, na ocorrência de uma citopenia grave e dependência transfusional, evolução clonal ou evolução do IPSS. Para os grupos de IPSS intermediário 2 e alto risco, TCTH logo após o diagnóstico confere o melhor prognóstico. Para os pacientes sem doador aparentado HLA-idêntico, o TCTH de doadores não aparentados pode ser uma alternativa e deve ser individualizado diante da perspectiva de cura, limitada, entretanto, pela elevada morbidade e mortalidade associada a este procedimento. O papel do TCTH autólogo e do TCTH com condicionamento não mieloablativo para os pacientes com diagnóstico de SMD precisa ser melhor definido.

Hematopoietic cell transplantation (HCT) is a curative therapy for patients with a diagnosis of myelodysplastic syndromes (MDS). The indications and the timing of HCT for MDS have remained controversial due to the high risk of morbidity and mortality related to this procedure. In view of the current evidence, for patients with an HLA-matched related donor with low and intermediate 1 IPSS, the best strategy is to delay transplantation, but to perform it prior to the development of acute myelogenous leukemia. For patients with intermediate 2 and high risk IPSS groups, transplantation soon after diagnosis confers the best prognosis. For patients lacking a HLA-matched related donor, studies involving a large number of patients with a long-term follow-up are needed to define the best option among conventional chemotherapy, autologous HCT or HCT from unrelated donors. For patients who are not candidates for myeloablative conditioning, prospective randomized trials are needed to define if non-myeloablative conditioning reduces the mortality not related to relapse without increasing the probability of relapse.