Navegando por Palavras-chave "Multiple myeloma"

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    Analysis of polymorphism at site -174 G/C of interleukin-6 promoter region in multiple myeloma
    (Associação Brasileira de Divulgação Científica, 2007-02-01) Duch, Cibele Repele [UNIFESP]; Figueiredo, Maria Stella [UNIFESP]; Ribas, Christian [UNIFESP]; Almeida, Manuella de Souza Sampaio [UNIFESP]; Colleoni, Gisele Wally Braga [UNIFESP]; Bordin, Jose Orlando [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    It is well established that interleukin-6 (IL-6) is an essential growth factor for multiple myeloma (MM) and patients with increased IL-6 levels have a poor prognosis. In healthy subjects, the presence of the C allele at a polymorphic site (-174 G/C) of the IL-6 gene is related to low IL-6 levels. In view of the potential association of this particular polymorphism with IL-6 concentration, and the relevance of IL-6 in MM pathogenesis, the objective of the present study was to investigate the prevalence of IL-6 (-174 G/C) promoter polymorphism and its association with development of MM in Brazilian individuals. We investigated the prevalence of these alleles in 52 patients and 60 healthy subjects (matched by age, sex, and race) of a Brazilian population. Thirty patients were male (42.4%), 24 (46.2%) were white and the median age at diagnosis was 58.5 years (range: 28 to 84 years). To determine the IL-6 (-174 G/C) polymorphism, molecular analysis was performed by polymerase chain reaction followed by endonuclease restriction digestion. The genotype distributions observed in the group of patients were 4% CC, 42% GC and 54% GG. The C allele frequency was 0.25. These results were similar to the control group, suggesting no impact of this polymorphism on the susceptibility to MM.
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    ANKHD1 regulates cell cycle progression and proliferation in multiple myeloma cells
    (Elsevier B.V., 2012-12-14) Dhyani, Anamika; Duarte, Adriana S. S.; Machado-Neto, Joao A.; Favaro, Patricia [UNIFESP]; Ortega, Manoela Marques; Saad, Sara T. Olalla; Universidade Estadual de Campinas (UNICAMP); Universidade Federal de São Paulo (UNIFESP)
    ANKHD1 is a multiple ankyrin repeat containing protein, highly expressed in cancers, such as acute leukemia. the present study was undertaken to determine the expression and functional significance of ANKHD1 in human Multiple Myeloma (MM). We found that ANKHD1 is highly expressed in MM patient cells and cell lines. in vitro, lentiviral mediated ANKHD1-shRNA inhibited proliferation and delayed S to G2M cell cycle progression in glucocorticoid resistant (U266) and sensitive (MM1S) MM cells. Further ANKHD1 silencing resulted in upregulation of cyclin dependent kinase inhibitor p21 irrespective of the p53 status of the MM cell lines. These data suggest that ANKHD1 might have a role in MM cell proliferation and cell cycle progression by regulating expression of p21. (C) 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
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    ANKHD1 represses p21 (WAF1/CIP1) promoter and promotes multiple myeloma cell growth
    (Elsevier B.V., 2015-01-01) Dhyani, Anamika; Machado-Neto, Joao A.; Favaro, Patricia [UNIFESP]; Olalla Saad, Sara T.; Universidade Estadual de Campinas (UNICAMP); Universidade Federal de São Paulo (UNIFESP)
    ANKHD1 (Ankyrin repeat and KH domain-containing protein 1) is highly expressed and plays an important role in the proliferation and cell cycle progression of multiple myeloma (MM) cells. ANKHD1 downregulation modulates cell cycle gene expression and upregulates p21 irrespective of the TP53 mutational status of MM cell lines. the present study was aimed to investigate the role of ANKHD1 in MM in vitro clonogenicity and in vivo tumourigenicity, as well as the role of ANKHD1 in p21 transcriptional regulation. ANKHD1 silencing in MM cells resulted in significantly low no. of colonies formed and in slow migration as compared to control cells (p < 0.05). Furthermore, in xenograft MM mice models, tumour growth was visibly suppressed in mice injected with ANKHD1 silenced cells compared to the control group. There was a significant decrease in tumour volume (p = 0.006) as well as in weight (p = 0.02) in the group injected with silenced cells compared to those of the control group. Co-immunoprecipitation and chromatin immunoprecipitation (ChIP) assays confirmed the interaction between p21 and ANKHD1. Moreover, overexpression of ANKHD1 downregulated the activity of a p21 promoter in luciferase assays. Decrease in luciferase activity suggests a direct role of ANKHD1 in p21 transcriptional regulation. in addition confocal analysis after U266 cells were treated with Leptomycin B (LMB) for 24 h showed accumulation of ANKHD1 inside the nucleus as compared to untreated cells where ANKHD1 was found to be predominantly in cytoplasm. This suggests ANKHD1 might be shuttling between cytoplasm and nucleus. in conclusion, ANKHD1 promotes MM growth by repressing p21 a potent cell cycle regulator. (C) 2014 Elsevier B.V. All rights reserved.
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    Anti-myeloma effects of ruxolitinib combined with bortezomib and lenalidomide: A rationale for JAK/STAT pathway inhibition in myeloma patients
    (Elsevier Ireland Ltd, 2017) de Oliveira, Mariana B. [UNIFESP]; Fook-Alves, Veruska L. [UNIFESP]; Eugenio, Angela I. P. [UNIFESP]; Fernando, Rodrigo C. [UNIFESP]; Sanson, Luiz Felipe G. [UNIFESP]; de Carvalho, Mariana F. [UNIFESP]; Braga, Walter M. T. [UNIFESP]; Davies, Faith E.; Colleoni, Gisele W. B. [UNIFESP]
    JAK proteins have been linked with survival and proliferation of multiple myeloma (MM) cells
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    Caracterização imunofenotípica das células plasmáticas em pacientes portadores de mieloma múltiplo
    (Sociedade Brasileira de Patologia ClínicaSociedade Brasileira de PatologiaSociedade Brasileira de Citopatologia, 2010-08-01) Leite, Luiz Arthur Calheiros [UNIFESP]; Almeida, Manuela S.; Kimura, Elisa S. [UNIFESP]; Bigonha, Jandey G. [UNIFESP]; Colleoni, Gisele Wally Braga [UNIFESP]; Chauffaille, Maria de Lourdes Lopes Ferrari [UNIFESP]; Yamamoto, Mihoko [UNIFESP]; Universidade Federal de Pernambuco; Centro de Estudos Superiores de Maceió Faculdade de Ciências Biológicas e da Saúde; Universidade Federal de São Paulo (UNIFESP)
    INTRODUCTION AND OBJECTIVE: Multiple myeloma is an incurable malignancy characterized by the proliferation of a single clone of plasma cells in bone marrow. The aim of this study was to evaluate the frequency and prognostic value of the expression of aberrant phenotypes in patients with multiple myeloma by multiparametric flow cytometry. METHODS: The study was carried out at Department of Hematology and Hemotherapy of Federal University of São Paulo and 30 patients with MM were analyzed prospectively. In an attempt to identify myeloma cells by flow cytometry (FACSCalibur, BD), specific monoclonal antibodies anti-CD138, anti-CD38 and anti-CD45 were used for the selection of plasma cells. The control group comprised four healthy bone marrow donors. RESULTS: All myeloma plasma cells expressed at least one aberrant phenotype and CD56+++, CD117++, CD33++, CD13++ and CD28++ markers were more frequently observed in 88% of patients. Lymphoid markers were found in cases with a higher number of aberrant phenotypes. DISCUSSION: CD56+++ and CD28++ antigens showed high levels of β2-microglobulin, which are associated with more aggressive stages of the disease and larger tumor mass. The absence of adhesion molecule CD56 was associated with high levels of β2M and calcium ion, showing that this finding may have prognostic value. CONCLUSION: From this study it was concluded that the aberrant phenotypes are present in most cases of MM, and immunophenotyping by multiparametric flow cytometry is a useful tool to distinguish normal plasma cells from myeloma plasma cells.
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    Caracterização imunofenotípica, do perfil de expressão gênica e funcional de células-tronco mesenquimais derivadas de medula óssea de pacientes com mieloma múltiplo
    (Universidade Federal de São Paulo (UNIFESP), 2017-11-21) Fernando, Rodrigo Carlini [UNIFESP]; Colleoni, Gisele Wally Braga [UNIFESP]; http://lattes.cnpq.br/2918635854077904; http://lattes.cnpq.br/0799332858520582; Universidade Federal de São Paulo (UNIFESP)
    Introdução: Evidências crescentes sugerem que o microambiente tumoral possui papel chave na manutenção e progressão de vários tipos de câncer, incluindo tumores sólidos e neoplasias malignas hematológicas. No mieloma múltiplo (MM), um câncer de células plasmáticas, a interação entre as células de MM e o ambiente circundante da medula óssea (MO), composto por elementos celulares e não celulares, promove a sobrevivência, o crescimento e a resistência a drogas das células tumorais. Entre os elementos celulares, as células-tronco mesenquimais da MO (do inglês, mesenchymal stem cells, MSC) merecem maior atenção devido ao seu papel emblemático na progressão da doença. Vários estudos tem mostrado diferenças entre as MSC de pacientes com MM (MM-MSC) e suas correspondentes normais (do inglês, normal donors MSC, ND-MSC), compreendendo desde alterações no perfil de expressão gênica e proteica, até alterações genéticas e funcionais. Objetivos: Os objetivos desse estudo foram caracterizar as diferenças nos perfis imunofenotípico, de expressão gênica e funcional entre as MM-MSC e as ND-MSC, com intuito de agregar novas informações à patogênese da doença e identificar potenciais alvos terapêuticos. Casuística e Métodos: Foram incluídos no estudo 19 casos de MM recém-diagnosticados, sem tratamento prévio, e sete doadores saudáveis de MO para transplante alogênico (não pareados por idade ou gênero). Os aspirados de MO foram coletados e processados para isolar as MSC, por separação magnética manual (CD105+). As MSC de ambos os grupos foram expandidas in vitro até atingir a quantidade de células necessária para os demais experimentos. As MSC foram caracterizadas por citometria de fluxo e por avaliação do potencial de diferenciação em osteoblastos. A análise do perfil de expressão gênica foi realizada por microarray, utilizando a plataforma GeneChip Human Exon 1.0 ST Array (Affymetrix). Após o pré-processamento dos dados brutos de microarray e identificação de genes diferencialmente expressos (GDE), foram realizadas análises de rede de co-expressão gênica e de enriquecimento funcional, utilizando ferramentas de bioinformática, a fim de identificar funções e vias biológicas enriquecidas. Por fim, com intuito de validar funcionalmente os achados das análises de bioinformática dos GDE, foi realizada a avaliação do ciclo celular, por citometria de fluxo, e a quantificação do comprimento telomérico dessas células por PCR em tempo real. Resultados: Não foram observadas diferenças imunofenotípicas, nem na capacidade de diferenciação em osteoblastos entre as células tumorais e normais. A análise de expressão gênica, por sua vez, mostrou 485 GDE nas MM-MSC, sendo 280 com expressão aumentada e 205 com expressão diminuída. As análises de bioinformática revelaram que as principais funções e vias biológicas enriquecidas, entre os genes com expressão diminuída, estavam relacionadas à progressão do ciclo celular e à ativação da resposta imune. Entretanto, não foram observadas diferenças estatisticamente significantes entre as distribuições dessas células nas diferentes fases do ciclo celular, nem no tamanho do comprimento telomérico das mesmas. Conclusão: Nossos resultados sugerem que as MM-MSC possuem perfil de expressão gênica distinto das ND-MSC, confirmando estudos anteriores. Dentre as funções desreguladas nas MM-MSC em decorrência da doença, destacam-se as alterações encontradas em genes responsáveis pela progressão do ciclo celular e pela ativação do sistema imune, os quais sugerem, respectivamente, que as MM-MSC são permanentemente dependentes das células de MM e que elas colaboram para a evasão do sistema imune por parte das células tumorais. Novas abordagens visando o último aspecto podem ser interessantes no tratamento do MM.
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    Efficacy and safety of bortezomib, thalidomide, and lenalidomide in multiple myeloma: An overview of systematic reviews with meta-analyses
    (Elsevier Science Inc, 2017) Aguiar, Patricia Melo; Lima, Tacio de Mendonca; Braga Colleoni, Gisele Wally [UNIFESP]; Storpirtis, Silvia
    This overview summarizes evidence for the efficacy and safety of bortezomib, thalidomide, and lenalidomide in patients with multiple myeloma. We searched the Medline, Scopus, and LILACS databases through August 2016, including systematic reviews with meta-analyses of randomized controlled trials assessing the efficacy and/or safety of bortezomib, thalidomide, or lenalidomide in patients with multiple myeloma. Two authors performed study selection, data extraction, and quality assessment using AMSTAR and GRADE instruments. Twenty-nine studies satisfied the inclusion criteria. All three drugs significantly improved overall response and progression-free survival
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    Elevada incidência de anormalidades cromossômicas numéricas detectadas por FISH multicentromérico em pacientes com mieloma múltiplo
    (Sociedade Brasileira de Patologia ClínicaSociedade Brasileira de PatologiaSociedade Brasileira de Citopatologia, 2007-02-01) Chauffaille, Maria de Lourdes Lopes Ferrari [UNIFESP]; Ribeiro Jr., Aníbal [UNIFESP]; Yamamoto, Mihoko [UNIFESP]; Rodrigues, Maria Madalena; Almeida, Manuella S. S. [UNIFESP]; Ribas, Christian [UNIFESP]; Calheiros, Luis A. [UNIFESP]; Colleoni, Gisele Wally Braga [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    This study aimed to characterize genetic alterations by interphase multicentromeric FISH focusing on chromosomal numerical abnormalities and using some locus specific probes for the most frequent aberrations found in the disease, in a homogeneous cohort of 34 advanced stage, but recently diagnosed MM patients; 97% had numerical chromosomal abnormalities detected by FISH, being 75% hyperdiploid, 18% hypodiploid and 3% tri/tetraploid. Using locus specific probes, we found 13q deletion in 30% and IGH rearrangement in 25% of cases. Grouping hypodiploid patients together with del13q (unfavorable group) and comparing them to the remaining cases (non unfavorable group) we found a trend towards younger patients presenting more unfavorable abnormalities (p = 0.06) and significant lower hemoglobin level (Hb < 8.5 mg/dl, p = 0.03).
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    Expression of adhesion molecules on CD34+ cells from steady-state bone marrow before and after mobilization and their association with the yield of CD34+ cells
    (Korean Soc Hematology, 2018) Cecyn, Karin Zattar [UNIFESP]; Kimura, Eliza Yuriko Sugano [UNIFESP]; Lima, Dulce Marta S. M. [UNIFESP]; Yamamoto, Miyoko [UNIFESP]; Bordin, Jose Orlando [UNIFESP]; Oliveira, José Salvador Rodrigues de [UNIFESP]
    Background Cell adhesion molecules (CAMs) expressed on hematopoietic progenitor cells (HPCs), endothelial cells, and stromal cells play a pivotal role in the mobilization of CD34+ cells. Herein, we conducted a non-randomized peripheral blood stem cell (PBSC) mobilization study aimed to compare the potential differences in the expressions of several CAMs and chemokines on CD34+ cells obtained from bone marrow aspirate before and after HPC mobilization from patients with hematologic malignancies and healthy donors. Methods Three-color cytofluorometric analysis was used to compare the expressions of CAMs and chemokines in the bone marrow before and after mobilization. Results For all studied groups, CAM expression among those with good and poor yields of CD34+ cells was significantly correlated with VCAM-1 (P=0.007), CD44 (P=0.027), and VLA-4 (P=0.014) expressions. VCAM-1 (P=0.001), FLT-3 (P=0.001), CD44 (P=0.011), VLA-4 (P=0.001), and LFA-1 (P=0,001) expressions were higher before HPC mobilization than after HPC mobilization. By contrast, the expression of CXCR4 significantly varied before and after mobilization only among those with successful PBSC mobilization (P=0.002). Conclusion We attempted to identify particular aspects of CAMs involved in CD34+ cell mobilization, which is a highly complex mechanism that involves adhesion molecules and matrix metalloproteases. The mechanism by which CD34+ cell mobilization is activated through proteolytic enzymes is not fully understood. We believe that CXCR4, VLA-4, CD44, and VCAM-1 are the most important molecules implicated in HPC mobilization, particularly because they show a correlation with the yield of CD34+ cells collected via large volume leukapheresis.
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    Expression of eight genes of nuclear factor-kappa B pathway in multiple myeloma using bone marrow aspirates obtained at diagnosis
    (F Hernandez, 2009-08-01) Almeida, Manuella de Souza Sampaio [UNIFESP]; Vettore, Andre Luiz [UNIFESP]; Yamamoto, Mihoko [UNIFESP]; Chauffaille, Maria de Lourdes Lopes Ferrari [UNIFESP]; Zago, Marco Antonio; Colleoni, Gisele Wally Braga [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Ludwig Inst Canc Res; Universidade de São Paulo (USP)
    Purpose: To evaluate the expression of NF-kappa B pathway genes in total bone marrow samples obtained from MM at diagnosis using real-time quantitative PCR and to evaluate its possible correlation with disease clinical features and survival. Material and methods: Expression of eight genes related to NF-kappa B pathway (NFKB1, IKB, RANK, RANKL, OPG, IL6, VCAM1 and ICAM1) were studied in 53 bone marrow samples from newly diagnosed MM patients and in seven normal controls, using the Taqman system. Genes were considered overexpressed when tumor expression level was at least four times higher than that observed in normal samples. Results: The percentages of overexpression of the eight genes were: NFKB1 0%, IKB 22.6%, RANK 15.1%, RANKL 31.3%, OPG 7.5%, IL6 39.6%, VCAM1 10% and ICAM1 26%. We found association between IL6 expression level and International Staging System (ISS) (p = 0.01), meaning that MM patients with high ISS scores have more chance of overexpression of IL6. The mean value of ICAM1 relative expression was also associated with the ISS score (p = 0.02). Regarding OS, cases with IL6 overexpression present worse evolution than cases with IL6 normal expression (p = 0.04). Conclusion: We demonstrated that total bone marrow aspirates can be used as a source of material for gene expression studies in MM. In this context, we confirmed that IL6 overexpression was significantly associated with worse survival and we described that it is associated with high ISS scores. Also, ICAM1 was overexpressed in 26% of cases and its level was associated with ISS scores.
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    FOXP3 and CTLA4 overexpression in multiple myeloma bone marrow as a sign of accumulation of CD4(+) T regulatory cells
    (Springer, 2014-11-01) Tobias Braga, Walter Moises [UNIFESP]; Silva, Bruna Raphaeli da [UNIFESP]; Carvalho, Ana Carolina de [UNIFESP]; Maekawa, Yumi Hasegawa; Bortoluzzo, Adriana Bruscato; Rizzatti, Edgar Gil; Atanackovic, Djordje; Braga Colleoni, Gisele Wally [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Fleury Med & Saude; Insper Inst Educ & Res; Univ Med Ctr Hamburg Eppendorf
    Multiple myeloma (MM) development involves a series of genetic abnormalities and changes in the bone marrow (BM) microenvironment, favoring the growth of the tumor and failure of local immune control. T regulatory (Treg) cells play an important role in dampening anti-tumor immune responses while T-helper-17 (Th17) cells seem to be critical for the eradication of malignant cells. the aim of our study was to characterize the expression of Treg- and Th17-related genes in total myeloma BM samples to assess their role as biomarkers, prognostic factors, and possible therapeutic targets in this incurable disease.Expression of markers for Treg (FOXP3, CTLA4) and Th17 cells (ROR gamma t) was determined by quantitative real-time PCR in BM aspirates of 46 MM patients, four patients with monoclonal gammopathy of undetermined significance, five solitary plasmacytomas, and five healthy BM donors. Gene expression was evaluated regarding an influence on the patients' overall survival (OS).FOXP3 and CTLA4 presented a sixfold (p = 0.02) and 30-fold higher expression (p = 0.03), respectively, in MM patients than in controls. ROR gamma t expression was similar in MM patients and controls. Median OS of MM patients was 16.8 (range 4.5-29.1) months, and international staging system was the only independent prognostic factor for patients survival.Overexpression of FOXP3 and CTLA4 in total BM samples suggests a local accumulation of immunosuppressive Tregs, the MM tumor environment, possibly dampening anti-tumor host immune responses. Therapeutic approaches targeting Treg cells and restoring local anti-tumor immunity may provide new treatment strategies for this incurable malignancy.
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    Inibição da via JAK/STAT em linhagens celulares de mieloma múltiplo como potencial alvo terapêutico
    (Universidade Federal de São Paulo (UNIFESP), 2017-04-11) Oliveira, Mariana Bleker de [UNIFESP]; Colleoni, Gisele Wally Braga [UNIFESP]; http://lattes.cnpq.br/2918635854077904; http://lattes.cnpq.br/7224557267378627; Universidade Federal de São Paulo (UNIFESP)
    Introdução: A ativação constitutiva das proteínas JAK em mieloma múltiplo (MM) promove a sobrevivência e proliferação de células tumorais. O bloqueio das proteínas JAK pode contribuir para reduzir os efeitos auxiliares da sinalização aberrante nas células de mieloma e a inibição farmacológica das JAKs pode, portanto, ser uma estratégia terapêutica para o tratamento do mieloma. Objetivos: 1) Identificar, em linhagens celulares de MM, a existência de expressão relevante dos genes da via JAK/STAT JAK1 e JAK2 e validar os achados em amostras de pacientes recém-diagnosticados com MM; 2) Realizar estudos funcionais in vitro com as linhagens celulares de MM previamente selecionadas, em monocultura e co-cultura com células estromais normais, tratadas com um inibidor da via JAK/STAT (ruxolitinibe), associado a outras drogas utilizadas atualmente no tratamento de primeira linha do MM; 3) Avaliar a expressão global dos genes da via JAK/STAT e de proteínas de interesse em linhagens celulares de MM previamente selecionadas, selvagens e tratadas in vitro com o inibidor da via JAK/STAT. Métodos: A expressão dos genes JAK1 e JAK2 foi analisada por PCR em tempo real (qPCR). Para os experimentos in vitro, com ruxolitinibe, bortezomibe, e a combinação das duas drogas, as concentrações foram determinadas após verificação de qual dose é capaz de induzir pelo menos 50% de morte celular. A apoptose (marcação com anexina V e iodeto de propídeo) e o ciclo celular (marcação com iodeto de propídeo) foram analisados por citometria de fluxo. O PCR array para a via JAK/STAT foi avaliado em duplicatas com células tratadas com a combinação de ruxolitinibe e bortezomibe e não tratadas e o valor do fold change (2-ΔΔCt) foi calculado para todos os genes. As proteínas Beclina-1 (autofagia) e XBP-1s (UPR) foram escolhidas para avaliação por Western Blot. Resultados: A linhagem RPMI-8226 apresentou menor expressão do gene JAK1, quando comparada à linhagem U266, e não houve diferença estatisticamente significante para a expressão do gene JAK2. Dos 30 pacientes recém-diagnosticados com MM, 57% apresentaram hiperexpressão de JAK2 e 21%, de JAK1. Porém, a expressão de JAK1 e JAK2 não apresentou correlação com a sobrevida ou com as características clínico-laboratoriais dos pacientes. As concentrações utilizadas para a combinação das drogas foram: ruxolitinibe (30 μM para RPMI-8226 e 40 μM para U266) e bortezomibe (10 nM). Após 72 horas de tratamento com a combinação de ruxolitinibe e bortezomibe (R+B), as linhagens RPMI-8226 e U266 apresentaram 50% de células em apoptose tardia. A morte celular nas duas linhagens foi acompanhada por diminuição de expressão dos genes anti-apoptóticos após tratamento com R+B. A linhagem RPMI-8226 apresentou maior número de células na fase SubG0 (p<0,001) após tratamento com R+B e redução de células nas fases S (p<0,01) e G2/M (p<0,001). A linhagem U266 não apresentou alterações no ciclo celular, exceto por um pequeno aumento de células na fase SubG0 (p<0,05). A co-cultura com células estromais protegeu as células RPMI-8226 tratadas com combinação de R+B da apoptose, já que a porcentagem de células em apoptose caiu para 32% (p<0,001), em relação à monocultura. A adição da droga imunomoduladora lenalidomida reverteu o efeito protetor conferido pelas células estromais, já que a combinação de ruxolitinibe, bortezomibe e lenalidomida causou 62% de morte celular nas células tumorais (p<0,05). O tratamento em co-cultura com ruxolitinibe, bortezomibe e lenalidomida foi equivalente ao tratamento com bortezomibe, lenalidomida e dexametasona, combinação utilizada na prática clínica. O perfil de expressão dos genes da via JAK/STAT nas células tratadas com R+B, quando comparado ao perfil das respectivas linhagens selvagens, mostrou diminuição de expressão de genes das vias JAK/STAT, Ras/Raf/MAPK e PI3K/Akt/mTOR, principalmente na linhagem RPMI-8226, com mudanças insignificantes no padrão de expressão da linhagem U266. Após tratamento com R+B, houve aumento na expressão proteica de XBP-1s. Conclusão: Neste trabalho, demonstramos que existe aumento de expressão do gene JAK2 em quase 60% dos pacientes com MM, além dos efeitos do inibidor de JAK1/2, ruxolitinibe, na apoptose tardia, ciclo celular e expressão de genes da via JAK/STAT, em pelo menos uma das linhagens celulares de MM. Nesse cenário, a via JAK/STAT pode configurar como um alvo terapêutico a ser explorado, já que se encontra constitutivamente ativa em uma porcentagem relevante de pacientes e contribui para a sobrevivência de células tumorais do MM.
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    Multiple myeloma with cells typically seen in storage diseases
    (Associação Brasileira de Hematologia e Hemoterapia e Terapia Celular, 2011-02-01) Boturão-Neto, Edmir [UNIFESP]; Yamamoto, Mihoko [UNIFESP]; Menezes, Yara; Bordin, Jose Orlando [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Hospital Alemão Oswaldo Cruz CICAP Laboratory
    We report on a rare case of multiple myeloma with atypically large cells containing a great amount of azurophilic inclusions usually seen in storage diseases.
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    Number of expressed cancer/testis antigens identifies focal adhesion pathway genes as possible targets for multiple myeloma therapy
    (Taylor & Francis Ltd, 2010-08-01) Andrade, Valeria C. C.; Vettore, Andre L. [UNIFESP]; Panepucci, Rodrigo A.; Almeida, Manuella S. S.; Yamamoto, Mihoko; De Carvalho, Fabricio; Caballero, Otavia L.; Zago, Marco Antonio; Colleoni, Gisele W. B. [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Universidade de São Paulo (USP); Ludwig Inst Canc Res
    Considering that the importance of cancer/testis (CT) antigens in multiple myeloma (MM) biology is still under investigation, the present study aimed to: (1) identify genes differentially expressed in MM using microarray analysis of plasma cell samples, separated according to the number of expressed CTs; (2) examine possible pathways related to MM pathogenesis; (3) validate the expression of candidate genes by quantitative real-time PCR (RQ-PCR). Three samples predominantly positive (>6 expressed), including the U266 cell line, and three samples predominantly negative (0 or 1 expressed CT for the 13 analyzed CT antigens), were submitted for microarray analysis. Validation by RQ-PCR from 24 MM samples showed that the ITGAS gene was downregulated in predominantly positive (>6 expressed CTs, p = 0.0030) and in tumor versus normal plasma cells (p = 0.0182). the RhoD gene was overexpressed in tumor plasma cells when compared to normal plasma cells (p = 0.0339). Results of the microarray analysis corroborate the hypothesis that MM could be separated into predominantly positive and predominantly negative expression. the differential expression of ITGA5 and RhoD suggests disruption of the focal adhesion pathway in MM and offers a new target field to be explored in this disease.
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    Papel da família da heat shock protein 70 (HSP70) como alvo terapêutico em mieloma múltiplo através de análises in vitro e in vivo
    (Universidade Federal de São Paulo (UNIFESP), 2017-03-07) Eugênio, Angela Isabel Pereira [UNIFESP]; Colleoni, Gisele Wally Braga [UNIFESP]; http://lattes.cnpq.br/2918635854077904; http://lattes.cnpq.br/1240012495336970; Universidade Federal de São Paulo (UNIFESP)
    Introduction: HSP70 has integrative role in protein degradation due to the interaction with many pathways, such as ubiquitin proteasome, unfolded protein response and autophagy. Objectives: To explore the role of HSP70 as a therapeutic target for multiple myeloma (MM) through in vitro and in vivo analyses. Methods: Bioluminescent cell lines RPMI8226-LUC-PURO and U266-LUC-PURO were treated with HSP70 inhibitor (VER155008) and proteasome inhibitor (bortezomib) for evaluation of apoptosis by flow cytometry. HSP70 members, unfolded protein response and autophagy genes were evaluated by qPCR in the same cell lines. Immunodeficient mice were used for subcutaneous xenograft model in two different approaches: RPMI8226-LUC-PURO cells into the right flank to induce a plasmocytoma. When the tumors became palpable mice were randomised to receive bortezomib, VER155008 or bortezomib plus VER155008 or no intervention. Histologic and protein expression analysis were performed. In a second model, each mouse was inoculated at the same moment, with RPMI8226-LUC-PURO cells into the left and U266-LUC-PURO into the righ flank. Mice were randomised into four groups of treatment and received intravenously proteassome and HSP70 inhitors immediately after xenotransplantation of the cell lines. Bioluminescence (BLI) was measured once a day for seven days. Results: RPMI8226- LUC-PURO and U266-LUC-PURO cell lines express HSP70 family genes, XBP-1 and BECLINA-1. Both cell lines treated with bortezomib, VER155008 (50μM and 80μM), isolated or combined, responded with increased expression of HSPA1A/HSPA1B. RPMI8226-LUC-PURO also showed increased expression of HSPA5 and XBP-1 genes after treatment with VER155008 (50μM). RPMI8226-LUC-PURO almost 60% of late apoptosis after treatment with bortezomib (100nM) alone. Treatment with VER155008, isolated or combined with bortezomib, did not add benefit to bortezomib treatment in 128 this cell line. However, U266-LUC-PURO cell line showed over 60% of cell death after treatment with VER155008 (80μM) alone and also with VER155008 (80μM) plus bortezomib, 48 hours after treatmet. In vivo data showed tumor growth reduction by bioluminescence in the group with RPMI8226-LUC-PURO plasmocytoma after treatment with bortezomib, VER155008 or combination of drugs compared to the control group. Nevertheless, the expression of HSP70 proteins, XBP-1s and BECLINA-1 had no statistically significant changes, except for reduction of XBP-1s protein relative expression in to tumors treated with VER155008. For the group of mice that had no prior induction of tumor, treatment with bortezomib, isolated or combined with VER155008 showed inhibition of tumor growth (or incipient growth) assessed by bioluminescence after one week for both cell lines when compared with the control group. Conclusion: Our study shows that the combination of proteasome and HSP70 inhibitors induced cell death in tumor cells in vitro and in vivo (late apoptosis induction and inhibition of tumor growth). Since HSP70 connects multiple signaling pathways that work synergistically to protect tumor cells from death by proteotoxic stress, it can represent a key role to establish a new approach for the treatment of MM (after achievement of the best response or as maintanence therapy) mostly in patients with deletion of 17p (like U266 cell line).
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    Plasmablastic multiple myeloma is associated with increased vascular endothelial growth factor immunoexpression
    (Associação Brasileira de Divulgação Científica, 2005-11-01) Ribas, C. [UNIFESP]; Colleoni, Gisele Wally Braga [UNIFESP]; Almeida, M.s.s. [UNIFESP]; Aguiar, K.c.c. [UNIFESP]; Silva, M.r.r. [UNIFESP]; Bordin, Jose Orlando [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    The biologic basis of the negative prognosis of plasmablastic myeloma is not fully understood. To determine whether histologically aggressive multiple myeloma (MM) is associated with a more angiogenic marrow environment, bone marrow samples from 50 recently diagnosed MM patients were evaluated. Twelve percent (6/50) of patients presented plasmablastic MM, and this feature correlated with moderate/strong intensity of vascular endothelial growth factor staining of plasma cells (P = 0.036). Although plasmablastic MM was not associated with increasing of microvessel density, this new evidence of increased expression of vascular endothelial growth factor on plasmablasts suggests that the adverse prognosis conferred by plasmablastic disease may be due, at least in part, to secretion of this angiogenic cytokine, also suggesting that the subset of MM patients with plasmablastic features may derive particular benefit from antiangiogenic therapies.
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    Primary systemic amyloidosis associated with multiple myeloma
    (Sociedade Brasileira de Dermatologia, 2012-02-01) Oliveira, Ederson Valei Lopes De; Pozetti, Ana Carolina Garcia; Pozetti, Eurides Maria De Oliveira; Antonio, João Roberto; Michalany, Nilceo Schwery [UNIFESP]; Faculdade de Medicina de São José do Rio Preto; Universidade Federal de São Paulo (UNIFESP)
    This case report is about a 48-year-old female patient with systemic amyloidosis and multiple myeloma simultaneously. Amyloid cutaneous infiltrative lesions like papules, nodules, or plaques with a serous-hemorrhagic aspect were found in the eyelids, neck and retroauricular region, among others. She had presented intermittent papular lesions on the upper eyelids one year before, which worsened following local trauma. A local skin biopsy showed amorphous and eosinophilic substance in the dermis. Congo red staining confirmed the amyloid deposits. Abnormal exams: proteinuria (570mg/24h), Bence-Jones proteinuria and clonal plasma cells (70%) found in myelogram. Following the diagnosis of multiple myeloma based on amyloid skin lesions, the patient was referred to the Hematology service and died 5 months after the diagnosis.
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    The renal and hepatic distribution of Bence Jones proteins depends on glycosylation: A scintigraphic study in rats
    (Assoc Bras Divulg Cientifica, 1997-07-01) Prado, Maria José Brandão de Almeida [UNIFESP]; Nicastri, Ana Lucia; Costa, P. L. A.; Rockman, T.; Tersariol, Ivarne Luis dos Santos [UNIFESP]; Nader, Helena Bonciani [UNIFESP]; Barros, Rubens Toledo; Prado, Euthymia Brandão de Almeida; Universidade de São Paulo (USP); Universidade Federal de São Paulo (UNIFESP)
    The aim of the present study was to evaluate renal and liver distribution of two monoclonal immunoglobulin light chains. the chains were purified individually from the urine of patients with multiple myeloma and characterized as lambda light chains with a molecular mass of 28 kDa. They were named BJg (high amount of galactose residues exposed) and BJs (sialic acid residues exposed) on the basis of carbohydrate content. A scintigraphic study was performed on male Wistar rats weighing 250 g for 60 min after iv administration of 1 mg of each protein (7.4 MBq), as the intact proteins and also after carbohydrate oxidation. Images were obtained with a Siemens gamma camera with a high-resolution collimator and processed with a MicroDelta system. Hepatic and renal distribution were established and are reported as percent of injected dose. Liver uptake of BJg was significantly higher than liver uptake of BJs (94.3 vs 81.4%) (P<0.05). This contributed to its greater removal from the intravascular compartment, and consequently lower kidney accumulation of BJg in comparison to BJs (5.7 vs 18.6%) (P<0.05). After carbohydrate oxidation, there was a decrease in hepatic accumulation of both proteins and consequently a higher renal overload. the tissue distribution of periodate-treated BJg was similar to that of native BJs: 82.7 vs 81.4% in the liver and 17.3 vs 18.6% in the kidneys. These observations indicate the important role of sugar residues of Bence Jones proteins for their recognition by specific membrane receptors, which leads to differential tissue accumulation and possible toxicity.
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    SAGE analysis highlights the importance of p53csv, ddx5, mapkapk2 and ranbp2 to multiple myeloma tumorigenesis
    (Elsevier B.V., 2009-06-08) Felix, Roberta S. [UNIFESP]; Colleoni, Gisele W. B. [UNIFESP]; Caballero, Otavia L.; Yamamoto, Mihoko [UNIFESP]; Almeida, Manuella S. S. [UNIFESP]; Andrade, Valeria C. C. [UNIFESP]; Chauffaille, Maria de Lourdes L. F. [UNIFESP]; Silva, Wilson A. da; Begnami, Maria Dirlei; Soares, Fernando Augusto; Simpson, Andrew J.; Zago, Marco Antonio; Vettore, Andre L. [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Ludwig Inst Canc Res; Universidade de São Paulo (USP); Fundacao Antonio Prudente
    Serial analysis of gene expression (SAGE) allows a comprehensive profiling of gene expression within a given tissue and also an assessment of transcript abundance. We generated SAGE libraries from normal and neoplastic plasma cells to identify genes differentially expressed in multiple myeloma (MM). Normal plasma cells were obtained from palatine tonsils and MM SAGE library was generated from bone marrow plasma cells of MM patients. We obtained 29,918 SAGE tags from normal and 10,340 tags from tumor libraries. Computer-gene rated genomic analysis identified 46 upregulated genes in the MM library. Ten upregulated genes were selected for further investigation. Differential expression was validated by quantitative real-time PCR in purified plasma cells of 31 patients and three controls. P53CSV, DDX5, MAPKAPK2 and RANBP2 were found to be upregulated in at least 50% of the MM cases tested. All of them were also found upregulated in MM when compared to normal plasma cells in a meta-analysis using ONCOMINE microarray database. Antibodies specific to DDX5, RANBP2 and MAPKAPK2 were used in a TMA containing 57 MM cases and confirmed the expression of these proteins in 74%, 96%, and 21% of the MM samples, respectively. Analysis of differential expression using SAGE could identify genes important for myeloma tumorigenesis (P53CSV, DDX5, MAPKPK2 and RANBP2) and that could potentially be useful as therapeutic targets. (c) 2008 Elsevier Ireland Ltd. All rights reserved.
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    TGF beta R2 aberrant methylation is a potential prognostic marker and therapeutic target in multiple myeloma
    (Wiley-Blackwell, 2009-10-15) Carvalho, Fabricio de [UNIFESP]; Colleoni, Gisele Wally Braga [UNIFESP]; Almeida, Manuella de Souza Sampaio [UNIFESP]; Carvalho, Andre L.; Vettore, Andre Luiz [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Barretos Canc Hosp
    Multiple myeloma (MM) is an incurable hematological malignancy. Different studies demonstrated the occurrence of genetic and epigenetic alterations in MM. the aberrant methylation is one of the most frequent epigenetic alterations in human genome. This study evaluated the aberrant methylation status or 20 genes in 51 MM samples by quantitative methylation-specific PCR (QMSP) and compared the methylation profile with clinicopathological characteristics of the patients. the QMSP analyses showed that PTGS2 (100.0%), SFN (100.0%), CDKN2B (90.2%), CDH1 (88.2%), ESR1 (72.5%), HIC1 (70.5%), CCND2 (62.7%), DCC (45.1%) and TGF beta R2 (39.2%) are frequently hypermethylated in MM while aberrant methviation of RAR beta (16.6%), MGMT (12.5%), AIM1 (12.5%), CDKN2A (8.3%), SOCS1 (8.3%), CCNA1 (8.3%) and THBS1 (4.1%) are rare events. There was no methylation of GSTP1, MINT31, p14ARF and RB1 in the samples tested. Hypermethylation of ESR1 was correlated positively with isotype IgA, while aberrant methylation of THBS1 correlated negatively with isotype IgG. Furthermore, hypermethylation of DCC and TGF beta R2 were correlated with poor survival. the multivariate analysis showed ISS and TGF beta R2 hypermethylation strongly correlated with poor outcome. This study represents the first quantitative evaluation of promoter methylation in MM and our data provide evidence that TGF beta R2 hypermethylation, besides ISS, may be useful as prognostic indicator in this disease. (C) 2009 UICC