RESUMEN
BACKGROUND: Malignant pleural mesothelioma (MPM) a rare neoplasm linked to asbestos exposure is characterized by a poor prognosis. Soluble mesothelin is currently considered the most specific diagnostic biomarker. The aim of the study was to identify novel biomarkers by proteomic analysis of two MPM cell lines secretome. MATERIALS AND METHODS: The protein patterns of MPM cells secretome were examined and compared to a non-malignant mesothelial cell line using two-dimensional gel electrophoresis coupled to mass spectrometry. Serum levels of candidate biomarkers were determined in MPM patients and control subjects. RESULTS: Two up-regulated proteins involved in cancer biology, prosaposin and quiescin Q6 sulfhydryl oxidase 1, were considered candidate biomarkers. Serum levels of both proteins were significantly higher in MPM patients than control subjects. Combining the data of each receiver-operating characteristic analysis predicted a good diagnostic accuracy. CONCLUSION: A panel of the putative biomarkers represents a promising tool for MPM diagnosis.
Asunto(s)
Biomarcadores de Tumor/sangre , Neoplasias Pulmonares/sangre , Mesotelioma/sangre , Neoplasias Pleurales/sangre , Proteoma/metabolismo , Anciano , Estudios de Casos y Controles , Línea Celular Tumoral , Femenino , Proteínas Ligadas a GPI/sangre , Humanos , Neoplasias Pulmonares/patología , Masculino , Mesotelina , Mesotelioma/patología , Mesotelioma Maligno , Persona de Mediana Edad , Oxidorreductasas actuantes sobre Donantes de Grupos Sulfuro/sangre , Neoplasias Pleurales/patología , Curva ROC , Saposinas/sangre , Vías SecretorasRESUMEN
Saposin A is a post-translation product of the prosaposin (PSAP) gene that serves as an activator protein of the galactocerebrosidase (GALC) enzyme, and is necessary for the degradation of certain glycosphingolipids. Deficiency of saposin A leads to a clinical picture identical to that of early-infantile Krabbe disease caused by GALC enzyme deficiency. Galactosylsphingosine, also known as psychosine, is a substrate of the GALC enzyme that is known to be elevated in classic Krabbe disease. We present the case of an 18-month-old male with clinical and radiological findings concerning for Krabbe disease who had preserved GALC enzyme activity and negative GALC gene sequencing, but was found to have a homozygous variant, c.257â¯Tâ¯>â¯A (p.I86N), in the saposin A peptide of PSAP. Psychosine determination on dried blood spot at 18â¯months of age was elevated to 12â¯nmol/L (normal <3â¯nmol/L). We present this case to add to the literature on the rare diagnosis of atypical Krabbe disease due to saposin A deficiency, to report a novel presumed pathogenic variant within PSAP, and to suggest that individuals with saposin A deficiency may have elevated levels of psychosine, similar to children with classic Krabbe disease due to GALC deficiency.
Asunto(s)
Galactosilceramidasa/genética , Homocigoto , Leucodistrofia de Células Globoides/diagnóstico por imagen , Psicosina/sangre , Saposinas/deficiencia , Pruebas con Sangre Seca , Variación Genética , Humanos , Lactante , Leucodistrofia de Células Globoides/sangre , Leucodistrofia de Células Globoides/genética , Imagen por Resonancia Magnética , Masculino , Saposinas/sangre , Saposinas/genéticaRESUMEN
Patients with Gaucher disease (GD) have an increased risk of monoclonal gammopathies for which antigenic targets might play a role in their pathogenesis. Here we report the identification of saposin C (sapC) as high-titre (1:1 000 000) target structure of 7/16 GD-associated paraproteins. Anti-sapC immunoglobulin (Ig) showed identity with the paraprotein Ig type and subclass in each patient that showed anti-sapC immunoreactivity. Absorption and depletion studies completely removed the paraprotein from the sera of GD patients. No immunoreactivity against sapC was detected in healthy donors and in other plasma cell dyscrasias, demonstrating that anti-sapC reactivity is highly restricted to GD. Several uncharacterized forms of post-translational modified sapC were detected but their role in the pathogenesis is not clear. We confirm the frequent presence of low-titre (1:250) anti-lysolipid reactivities in the sera of GD patients but we could show that this immunoreactivity is not mediated by the paraprotein and is not restricted to GD patients.
Asunto(s)
Enfermedad de Gaucher/sangre , Gammopatía Monoclonal de Relevancia Indeterminada/sangre , Mieloma Múltiple/sangre , Paraproteínas/metabolismo , Saposinas/sangre , Femenino , Humanos , MasculinoRESUMEN
BACKGROUND: One major obstacle to schistosomiasis prevention and control is the lack of accurate and sensitive diagnostic approaches, which are essential for planning, targeting, and evaluating disease control efforts. METHODS: Based on bioinformatics analysis, we identified a multigene family of saposin-like protein (SAPLP) in the schistosome genomes. Schistosoma japonicum SAPLPs (SjSAPLPs), including recently reported promising biomarker SjSP-13, were systematically and comparatively assessed as immunodiagnostic antigens for schistosomiasis japonica. RESULTS: Two novel antigens (SjSAPLP4 and SjSAPLP5) could specifically react to serum samples from both S. japonicum-infected laboratory animals and patients. The sensitivities of SjSAPLP4, SjSAPLP5, and SjSP-13 for immunodiagnosis were 98% (95% confidence interval, 88.0%-99.9%), 96% (85.1%-99.3%), and 88% (75.0%-95.0%), respectively, and 100% (91.1%-100%) specificity was observed for the 3 antigens with enzyme-linked immunosorbent assay; there was no cross-reaction with clonorchiosis (0 of 19 patients), echinococcosis (0 of 20 patients), or trichinellosis (0 of 18 patients) for the 3 antigens. Antibodies to the 3 antigens could be detected in the serum samples of rabbits infected with 1000 cercariae as early as 3-4 weeks after infection. CONCLUSIONS: These results suggest that SjSAPLP4 and SjSAPLP5 could serve as novel biomarkers for the immunodiagnosis of schistosomiasis japonica, which will further improve diagnostic sensitivity and specificity.
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Biomarcadores/sangre , Familia de Multigenes/genética , Saposinas/sangre , Saposinas/inmunología , Esquistosomiasis Japónica/diagnóstico , Esquistosomiasis Japónica/inmunología , Animales , Anticuerpos Antihelmínticos/inmunología , Antígenos Helmínticos/inmunología , Femenino , Humanos , Pruebas Inmunológicas/métodos , Masculino , Ratones , Ratones Endogámicos BALB C , Familia de Multigenes/inmunología , Conejos , Schistosoma japonicum/genética , Esquistosomiasis Japónica/sangre , Esquistosomiasis Japónica/parasitología , Sensibilidad y EspecificidadRESUMEN
Prosaposin (PSAP) deficiency is an ultra-rare, fatal infantile lysosomal storage disorder (LSD) caused by variants in the PSAP gene, with seven subjects reported so far. Here, we provide the clinical, biochemical and molecular characterization of two additional PSAP deficiency cases. Lysoplex, a targeted resequencing approach was utilized to identify the variant in the first patient, while quantification of plasma lysosphingolipids (lysoSLs), assessed by liquid chromatography mass spectrometry (LC-MS/MS) and brain magnetic resonance imaging (MRI), followed by Sanger sequencing allowed to attain diagnosis in the second case. Functional studies were carried out on patients' fibroblast lines to explore the functional impact of variants. The two patients were homozygous for two different truncating PSAP mutations (c.895G>T, p.Glu299*; c.834_835delGA, p.Glu278Aspfs*27). Both variants led to a complete lack of processed transcript. LC-MS/MS and brain MRI analyses consistently provided a distinctive profile in the two children. Quantification of specific plasma lysoSLs revealed elevated levels of globotriaosylsphingosine (LysoGb3) and glucosylsphingosine (GlSph), and accumulation of autophagosomes, due to a decreased autophagic flux, was observed. This report documents the successful use of plasma lysoSLs profiling in the PSAP deficiency diagnosis, as a reliable and informative tool to obtain a preliminary information in infantile cases with complex traits displaying severe neurological signs and visceral involvement.
Asunto(s)
Encéfalo/metabolismo , Leucodistrofia Metacromática/genética , Saposinas/deficiencia , Esfingolípidos/sangre , Encéfalo/diagnóstico por imagen , Encéfalo/patología , Cromatografía Liquida , Consanguinidad , Femenino , Humanos , Lactante , Leucodistrofia Metacromática/sangre , Leucodistrofia Metacromática/diagnóstico por imagen , Leucodistrofia Metacromática/patología , Imagen por Resonancia Magnética , Masculino , Mutación , Saposinas/sangre , Saposinas/genéticaRESUMEN
BACKGROUND: We previously cloned prosaposin (PSAP) from metastatic castrate-resistant prostate cancer (mCRPCa) cells and demonstrated its genomic amplification and/or overexpression in metastatic PCa cell lines, xenografts, and lymph node metastases. The clinicohistopathological significance of serum PSAP levels and its tissue expression and association with predictive or prognostic variable in primary or advanced PCa are not known. METHODS: We examined PSAP expression by immunohistochemical staining during early embryogenic development of the prostate and within a large tissue microarray which included 266 benign and malignant prostate tissues. In addition, serum PSAP levels in the age-adjusted normal male population and in 154 normal individuals and patients with primary or mCRPCa were measured by an ELISA assay. RESULTS: Univariate and multivariate analyses revealed a significant and inverse association between PSAP expression and clinical stages II and III tumors, dominant Gleason patterns 3 and 4, and seminal vesicle invasion. In the normal male population, the lowest serum PSAP level was detected before puberty, peaked at the most reproductive age group (20- to 39-year old), and then, decreased to a range between the two groups for men above 40-year old. Regardless of age and when compared with normal individuals, serum PSAP levels significantly decreased in primary organ-confined PCa, but increased in those with mCRPCa. CONCLUSION: Our results show that PSAP has the potential to differentiate between primary and advanced PCa. Additional large-scale studies are needed to define the usefulness of tissue expression or serum PSAP levels as a diagnostic or prognostic marker or as a therapeutic target in PCa.
Asunto(s)
Progresión de la Enfermedad , Neoplasias de la Próstata/sangre , Neoplasias de la Próstata/diagnóstico , Saposinas/sangre , Índice de Severidad de la Enfermedad , Adolescente , Adulto , Anciano , Envejecimiento/metabolismo , Apoptosis , Biomarcadores de Tumor/sangre , Estudios de Casos y Controles , Castración , Niño , Preescolar , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , Próstata/embriología , Próstata/metabolismo , Próstata/patología , Neoplasias de la Próstata/patología , Adulto JovenRESUMEN
OBJECTIVE: To evaluate the use of protein markers using immune-quantification assays and of metabolite markers using tandem mass spectrometry for the identification, at birth, of individuals who have a lysosomal storage disorder. METHODS: A retrospective analysis was conducted of Guthrie cards that were collected from newborns in Denmark during the period 1982-1997. Patients whose lysosomal storage disorder (LSD; 47 representing 12 disorders) was diagnosed in Denmark during the period 1982-1997 were selected, and their Guthrie cards were retrieved from storage. Control cards (227) were retrieved from the same period. Additional control cards (273) were collected from the South Australian Screening Centre (Australia). RESULTS: From 2 protein and 94 metabolite markers, 15 were selected and evaluated for their use in the identification of LSDs. Glycosphingolipid and oligosaccharide markers showed 100% sensitivity and specificity for the identification of Fabry disease, alpha-mannosidosis, mucopolysaccharidosis (MPS) IVA, MPS IIIA, Tay-Sachs disease, and I-cell disease. Lower sensitivities were observed for Gaucher disease and sialidosis. No useful markers were identified for Krabbe disease, MPS II, Pompe disease, and Sandhoff disease. The protein markers LAMP-1 and saposin C were not able to differentiate individuals who had an LSD from the control population. CONCLUSIONS: Newborn screening for selected LSDs is possible with current technology. However, additional development is required to provide a broad coverage of disorders in a single, viable program.
Asunto(s)
Biomarcadores/sangre , Enfermedades por Almacenamiento Lisosomal/diagnóstico , Tamizaje Neonatal/métodos , Anticuerpos Monoclonales , Antígenos CD/sangre , Antígenos CD/inmunología , Glicoesfingolípidos/sangre , Humanos , Recién Nacido , Proteínas de Membrana de los Lisosomas , Espectrometría de Masas/métodos , Oligosacáridos/sangre , Estudios Retrospectivos , Saposinas/sangre , Saposinas/inmunología , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: Fabry disease is an X-linked inborn error of glycosphingolipid catabolism resulting from a deficiency of the lysosomal exoglycohydrolase, alpha-galactosidase. Enzyme replacement therapy is currently available for Fabry disease, but early diagnosis before the onset of irreversible pathology will be mandatory for successful treatment. Presymptomatic detection would be possible through the use of a newborn-screening program. We report on the use of sensitive assays for the measurement of alpha-galactosidase protein and activity and for the protein saposin C, which are diagnostic markers for Fabry disease. METHODS: Two sensitive immunoassays for the measurement of alpha-galactosidase activity and protein were used to determine the concentrations of alpha-galactosidase in dried filter-paper blood spots and plasma samples from control patients and patients with a lysosomal storage disorder (LSD). RESULTS: Fabry hemizygous individuals were clearly identified from control populations by decreases in both alpha-galactosidase activity and protein. Fabry heterozygotes generally fell between the hemizygotes and controls. Including the measurement of saposin C enabled differentiation between Fabry heterozygotes and controls. In blood spots, all Fabry individuals could be distinguished from control blood spots as well as from 16 other LSD patients. CONCLUSIONS: The determination of alpha-galactosidase activity or protein in dried filter-paper blood spots could be used for the diagnosis of Fabry patients. With further validation, these assays could be used for the identification of Fabry patients in newborn-screening programs and may also be suitable for screening high-risk populations.