RESUMEN
BACKGROUND: Aldehyde dehydrogenase (ALDH) is highly active in physiological stem cells as well as in tumor-initiating cells of some malignancies including multiple myeloma (MM). Finding higher activity of ALDH in some cell subsets in monoclonal gammopathies (MG) could identify potential source of myeloma-initiating cells (MICs). METHODS: Bone marrow of 12 MM, 9 monoclonal gammopathy of undetermined significance (MGUS), and 10 healthy donors (HD) were analyzed by flow cytometry. ALDH activity of B-cells and plasma cells (PC) was analyzed using Aldefluor. RESULTS: Similar changes of ALDH activity were found during B-cell development in HD and MG. Decreasing of ALDH activity from immature to naïve B-cells was found. In postgerminal stages, the activity started to increase, and in PCs, the ALDH activity was the same as in immature B-cells. Increased ALDH activity of all PC subsets compared to naïve B-cells was found in MM as well as in HD, while in MGUS, only CD19- PCs have higher ALDH activity. In HD, ALDH activity was higher in CD19+ PCs compared with MG. CONCLUSIONS: Our results indicate that changes of ALDH activity are the natural phenomenon in B-cell development; thus, high ALDH activity as a single marker is not appropriate for MICs identification.
Asunto(s)
Aldehído Deshidrogenasa/metabolismo , Linfocitos B/enzimología , Paraproteinemias/diagnóstico , Paraproteinemias/enzimología , Células Plasmáticas/enzimología , Adulto , Anciano , Anciano de 80 o más Años , Antígenos de Superficie/metabolismo , Linfocitos B/patología , Biomarcadores , Médula Ósea/patología , Estudios de Casos y Controles , Activación Enzimática , Femenino , Humanos , Inmunofenotipificación , Masculino , Persona de Mediana Edad , Gammopatía Monoclonal de Relevancia Indeterminada/diagnóstico , Gammopatía Monoclonal de Relevancia Indeterminada/enzimología , Mieloma Múltiple/diagnóstico , Mieloma Múltiple/enzimología , Estadificación de Neoplasias , Células Plasmáticas/patologíaRESUMEN
AIMS: The study aimed at comparing two methods for evaluating thymidinekinase TK in serum - an older RIA method and novel DiviTum - in patients with MM and MGUS, and also comparing them with biochemical markers and degree of activity evaluated by imaging methods 99mTc-MIBI scintigraphy and 18F-FDG PET/CT. METHODS: Serum thymidinekinase TK levels were evaluated by DiviTum and an RIA method (TK REA kit by Immunotech);The study analyzed correlation of TK activity in serum with biochemical markers reflecting activity of MM: ß2-m, LDH, the ratio of kappa to lambda (κ/λ) free light chains and percentage of bone marrow plasma cells (BMPC). 99mTc-MIBI scintigraphy and 18F-FDG PET/CT were performed at the time of diagnosis. The degree of activity was expressed semiquantitatively. Scans were classified as 0 (normal activity), 1 (diffuse positivity) or 2 (focal positivity). RESULTS: We found a strong positive correlation between TK in serum evaluated by DiviTum and by TK REA.. The DiviTum analytic method extended the detection range and was able to detect higher levels of TK than the RIA method. DiviTum technique found positive correlation with ß2-m (r = 0.497) and LDH (r = 0.502) and moderate positive correlation with BMPC (r = 0.368). Significantly higher TK values measured by TK REA and DiviTum in the group of patients with MM (stages I, II or III) than in those with MGUS. Increased TK levels were observed in MIBI- or PET/CT-positive patients. Analysis of repeated measurements of TK in serum during treatment of MM patients found a correlation between change in TK measured by DiviTum and LDH during treatment. CONCLUSIONS: Analysis revealed a significant correlation between TK in serum and LDH, ß2-m and BMPC. Increased levels of TK in serum were observed in MIBI- or PET/CT-positive patients. Combination of positivity of imaging methods which can localize active tumor lesions and increased levels of TK in serum can have an impact on decision-making and optimization of the therapeutic approach.
Asunto(s)
Fluorodesoxiglucosa F18/farmacología , Gammopatía Monoclonal de Relevancia Indeterminada/enzimología , Mieloma Múltiple/enzimología , Tomografía de Emisión de Positrones , Timidina Quinasa/sangre , Tomografía Computarizada por Rayos X , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Gammopatía Monoclonal de Relevancia Indeterminada/diagnóstico por imagen , Mieloma Múltiple/diagnóstico por imagen , Pronóstico , Prohibitinas , Radioinmunoensayo , Radiofármacos/farmacologíaRESUMEN
BACKGROUND: Glutathione S-transferase P1 (GSTP1) is an important phase II enzyme involved in detoxification of carcinogens. GSTP1 gene overexpression has been observed in a variety of human cancers but there are no studies in plasma cell disorders. The aim of this study was to examine GSTP1 mRNA expression level in multiple myeloma (MM) and monoclonal gammopathy of undetermined significance (MGUS). In addition, we have determined GSTP1 polymorphic variants in order to estimate MM risk and their relationship with the expression level. Results were also correlated with laboratory parameters and clinical outcome. METHODS: Bone marrow mononuclear cells from 125 patients with plasma cell disorders were studied. Peripheral blood samples of 110 age and sex matched healthy controls were also evaluated. Real-Time Quantitative RT-PCR and PCR-RFLP assays were used. RESULTS: Upregulation of GSTP1 was observed in 37.7% MM and in 22.6% MGUS patients. A significant increase of GSTP1 expression in MM with respect to MGUS was detected (p=0.0427). Most MM patients that achieved complete remission had low transcription levels (77.8%) compared to those who did not reach this condition (44.4%) (p=0.0347). GSTP1 heterozygous carriers showed reduced expression compared to those with homozygous wild type genotype (p=0.0135). CONCLUSION: Our findings suggest, for the first time, a role for GSTP1 expression in development and/or progression of plasma cell disorders, and a probable influence of functional capacity of the enzyme on clinical outcome. These results and those of the literature support GSTP1 as an interesting tumor marker and a potential therapeutic target.
Asunto(s)
Gutatión-S-Transferasa pi/genética , Gammopatía Monoclonal de Relevancia Indeterminada/enzimología , Gammopatía Monoclonal de Relevancia Indeterminada/genética , Mieloma Múltiple/enzimología , Mieloma Múltiple/genética , ARN Mensajero/biosíntesis , Adulto , Anciano , Anciano de 80 o más Años , Alelos , Femenino , Regulación Enzimológica de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Genotipo , Gutatión-S-Transferasa pi/biosíntesis , Humanos , Masculino , Persona de Mediana Edad , ARN Mensajero/genética , Regulación hacia Arriba , Adulto JovenAsunto(s)
Terapia de Reemplazo Enzimático/métodos , Enfermedad de Gaucher/tratamiento farmacológico , Glucosilceramidasa/administración & dosificación , Linfoma de Células B/tratamiento farmacológico , Gammopatía Monoclonal de Relevancia Indeterminada/tratamiento farmacológico , 1-Desoxinojirimicina/administración & dosificación , 1-Desoxinojirimicina/análogos & derivados , 1-Desoxinojirimicina/uso terapéutico , Femenino , Enfermedad de Gaucher/complicaciones , Enfermedad de Gaucher/enzimología , Glucosilceramidasa/uso terapéutico , Humanos , Linfoma de Células B/complicaciones , Linfoma de Células B/enzimología , Persona de Mediana Edad , Gammopatía Monoclonal de Relevancia Indeterminada/complicaciones , Gammopatía Monoclonal de Relevancia Indeterminada/enzimología , EsplenectomíaRESUMEN
EGLN1 and EGLN3 are members of the egg-laying-defective 9 (EglN) prolyl-hydroxylases which during normoxia catalyse hydroxylation of the hypoxia-inducible factor (HIF)-1alpha, thereby promoting its ubiquitination by a complex containing the von Hippel-Lindau (VHL) tumour suppressor. EGLN3 also has pro-apoptotic activity in some cell types. Analyses of a well-characterised series of cases of plasma cell dyscrasias, including multiple myeloma (MM), Waldenström's macroglobulinaemia (WM) and monoclonal gammopathy of undetermined significance (MGUS) surprisingly demonstrated that the CpG island of EGLN3, and not EGLN1, is frequently methylated in these disorders. Multiple myeloma patients with a methylated EGLN3 promoter showed trends towards an increased risk of death, bone lytic lesions, anaemia, advanced stage of disease and the presence of extramedullary disease. Those individuals with methylation in the EGLN3 CpG island also had significantly lower albumin levels. These data suggest that the prolyl-hydroxylases may be a novel class of potential tumour suppressors in plasma cell neoplasia that warrant further investigation with regard to their potential utility as biomarkers. Moreover, we observed that EGLN3 is also methylated at high frequency in B-cell lymphoma subtypes, implying that loss of EGLN3 is an important epigenetic event not only in plasma cell neoplasias but also in B-cell neoplasias.