RESUMEN
PURPOSE: An efficient framework to identify disease-associated genes is needed to evaluate genomic data for both individuals with an unknown disease etiology and those undergoing genomic screening. Here, we propose a framework for gene selection used in genomic analyses, including applications limited to genes with strong or established evidence levels and applications including genes with less or emerging evidence of disease association. METHODS: We extracted genes with evidence for gene-disease association from the Human Gene Mutation Database, OMIM, and ClinVar to build a comprehensive gene list of 6,145 genes. Next, we applied stringent filters in conjunction with computationally curated evidence (DisGeNET) to create a restrictive list limited to 3,929 genes with stronger disease associations. RESULTS: When compared to manual gene curation efforts, including the Clinical Genome Resource, genes with strong or definitive disease associations are included in both gene lists at high percentages, while genes with limited evidence are largely removed. We further confirmed the utility of this approach in identifying pathogenic and likely pathogenic variants in 45 genomes. CONCLUSION: Our approach efficiently creates highly sensitive gene lists for genomic applications, while remaining dynamic and updatable, enabling time savings in genomic applications.
Asunto(s)
Genómica , Bases de Datos Factuales , Humanos , MutaciónRESUMEN
We have recently identified a small phosphoprotein, P20, as a common intracellular target for insulin and several of its antagonists, including amylin, epinephrine, and calcitonin gene-related peptide. These hormones elicit phosphorylation of P20 at its different sites, producing three phosphorylated isoforms: S1 with an isoelectric point (pI) value of 6.0, S2 with a pI value of 5.9, and S3 with a pI value of 5.6 (FEBS Letters 457:149-152 and 462:25-30, 1999). In the current study, we showed that P20 is one of the most abundant phosphoproteins in rat extensor digitorum longus (EDL) muscle. Insulin and amylin antagonize each other's actions in the phosphorylation of this protein in rat EDL muscle. Insulin inhibits amylin-evoked phosphorylation of S2 and S3, whereas amylin decreases insulin-induced phosphorylation of S1. In rats made insulin resistant by dexamethasone treatment, levels of the phosphoisoforms S2 and S3, which were barely detectable in healthy rats in the absence of hormone stimulation, were significantly increased. Moreover, the ability of insulin to inhibit amylin-evoked phosphorylation of these two isoforms was greatly attenuated. These results suggested that alterations in the phosphorylation of P20 might be associated with insulin resistance and that P20 could serve as a useful marker to dissect the cellular mechanisms of this disease.
Asunto(s)
Resistencia a la Insulina/fisiología , Proteínas Musculares/metabolismo , Músculo Esquelético/metabolismo , Fosfoproteínas/metabolismo , Tejido Adiposo/metabolismo , Amiloide/farmacología , Animales , Aorta/metabolismo , Péptido Relacionado con Gen de Calcitonina/farmacología , Línea Celular , Dexametasona/farmacología , Grasas de la Dieta/farmacología , Electroforesis en Gel Bidimensional , Electroforesis en Gel de Poliacrilamida , Epinefrina/farmacología , Proteínas del Choque Térmico HSP20 , Proteínas de Choque Térmico/metabolismo , Técnicas In Vitro , Insulina/farmacología , Polipéptido Amiloide de los Islotes Pancreáticos , Hígado/metabolismo , Masculino , Modelos Animales , Proteínas Musculares/genética , Proteínas Musculares/aislamiento & purificación , Músculo Esquelético/efectos de los fármacos , Músculo Liso Vascular/metabolismo , Miocardio/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/aislamiento & purificación , Fosforilación , Ratas , Ratas Wistar , Proteínas Recombinantes/metabolismo , Valores de Referencia , TransfecciónRESUMEN
Amylin, a 37-amino acid peptide, is cosecreted with insulin from the beta-cells of the pancreatic islets in normal response to physiological stimuli. It is the major protein of islet amyloid, which is usually present in the pancreases of people with non-insulin-dependent (type II) diabetes mellitus. Amylin elicits potent effects on carbohydrate metabolism in rodent tissues, causing insulin resistance in skeletal muscle and liver. A close structural relationship exists between amylin and the 2 calcitonin gene-related peptides, which are widely distributed neuropeptides and potent vasodilators. These exert biological effects similar to those of amylin on the organs primarily responsible for the regulation of carbohydrate metabolism. All 3 peptides are thought to cause their biological actions by binding to similar cell surface receptors. This article reviews the field of amylin and its role in the physiological regulation of carbohydrate metabolism, and in disease mechanisms associated with insulin resistance in diabetes mellitus, impaired glucose tolerance and essential hypertension. Potential therapeutic applications are also discussed.
Asunto(s)
Amiloide/farmacología , Amiloide/fisiología , Glucagón/metabolismo , Amiloide/antagonistas & inhibidores , Amiloide/metabolismo , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/fisiopatología , Humanos , Polipéptido Amiloide de los Islotes PancreáticosRESUMEN
Several reports have indicated that proteinase inhibitors can act as both stimulators and inhibitors of the growth of cultured cells. To confirm and extend these reports, we have purified the trypsin inhibitor from human urine (UTI). We have demonstrated a biphasic effect of this protein on the proliferation of human fibroblasts, and have identified two types of cellular binding site which may be responsible for the stimulatory and inhibitory aspects of this effect. Both aspects of UTI action have also been observed in human tumour cell cultures, but they are not consistently associated in these cells.
Asunto(s)
División Celular/efectos de los fármacos , Glicoproteínas/farmacología , Inhibidores de Tripsina/farmacología , Secuencia de Aminoácidos , Animales , Sitios de Unión , Bovinos , Células Cultivadas , ADN/biosíntesis , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Glicoproteínas/química , Glicoproteínas/aislamiento & purificación , Humanos , Mitógenos/farmacología , Datos de Secuencia Molecular , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/metabolismoRESUMEN
Inhibition of a cell-surface proteinase can inhibit the growth of many normal human cell types in culture. Some tumour cells are also sensitive to proteinase inhibitors, but others are resistant, and continue to grow in the presence of these inhibitors. Here we describe two human tumour cell lines which convert from the sensitive to the resistant state. In one case, the conversion occurs during routine passaging, but, in the other, it is determined by growth conditions, and is reversible.
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Proteínas de Unión al Calcio , Neoplasias del Colon/patología , Melanoma/patología , Proteínas de Transporte de Monosacáridos , Neoplasias/metabolismo , Proteínas de Unión Periplasmáticas , Células Tumorales Cultivadas/efectos de los fármacos , alfa 1-Antitripsina/farmacología , Proteínas Portadoras/farmacología , División Celular/efectos de los fármacos , Neoplasias del Colon/enzimología , Medios de Cultivo , Resistencia a Medicamentos , Endopeptidasas/metabolismo , Humanos , Hidrocortisona/farmacología , Insulina/farmacología , Melanoma/enzimología , Osteosarcoma/enzimología , Osteosarcoma/patología , Células Tumorales Cultivadas/citología , Células Tumorales Cultivadas/enzimologíaRESUMEN
The possibility that a growth-related proteinase may act by degrading a negative growth regulatory protein has been investigated. Proteinase inhibitors which inhibit the enzyme also enhance the accumulation of the growth regulator by human fibroblasts. The negative growth regulator shows a similar specificity of inhibition of cellular growth to inhibitors of the growth-related proteinase.
Asunto(s)
Proteínas de Unión al Calcio , Galactósidos/metabolismo , Sustancias de Crecimiento/aislamiento & purificación , Proteínas de Transporte de Monosacáridos , Proteínas de Unión Periplasmáticas , Inhibidores de Proteasas/farmacología , Animales , Sitios de Unión , Proteínas Portadoras/análisis , Bovinos , División Celular/efectos de los fármacos , Células Cultivadas , Cromatografía de Afinidad , Endotelio Corneal , Fibroblastos , Células HeLa , Humanos , Células Tumorales CultivadasRESUMEN
The proliferation of normal cells, and of early-passage cells from tumours, is inhibited as a consequence of inhibition of a cell-surface proteinase activity. In contrast, established tumour cell lines are usually resistant to these effects. It has previously been suggested that the change from sensitivity to resistance may be a consequence of tumour progression, but experiments with an additional three human tumour cell cultures show that this change is not inevitable.
Asunto(s)
Endopeptidasas/metabolismo , Secuencia de Aminoácidos , Carcinoma/enzimología , División Celular , Neoplasias del Colon/enzimología , Humanos , Datos de Secuencia Molecular , Células Tumorales Cultivadas , alfa 1-Antitripsina/farmacologíaRESUMEN
Human pancreatic secretory trypsin inhibitor inhibited cell-surface proteolytic activity in human fibroblasts. In the range of concentrations which caused proteinase inhibition, fibroblast proliferation was also inhibited by this reagent and by the ovine equivalent. At lower concentrations, there was some evidence for a mitogenic effect, and this was confirmed by obvious stimulation of DNA synthesis at these concentrations. Human alpha 1-proteinase inhibitor, previously demonstrated to be an inhibitor of fibroblast proliferation, was also mitogenic at concentrations lower than those which inhibited proteolytic activity and cell proliferation. Human pancreatic secretory trypsin inhibitor and epidermal growth factor apparently work through independent mechanisms, since their mitogenic effects are additive.
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ADN/biosíntesis , Mitógenos , Inhibidor de Tripsina Pancreática de Kazal/farmacología , Inhibidores de Tripsina/farmacología , alfa 1-Antitripsina/farmacología , Animales , División Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Factor de Crecimiento Epidérmico/farmacología , Fibroblastos/efectos de los fármacos , Humanos , OvinosRESUMEN
Previous experiments have shown that cultured human fibroblasts possess a cell-surface proteinase (the growth-related proteinase; GRP) which is essential to cell proliferation. In the present work, proteinase inhibition in defined and complex serum-free media and in pre-conditioned normal medium, still resulted in a corresponding inhibition of cell proliferation. Proteinase inhibition also blocked the action of a range of peptide growth factors and of a phorbol ester. Elevated extracellular calcium concentrations were still mitogenic in the presence of proteinase inhibitors. Proteinase inhibition did not affect the mobilisation of intracellular calcium, nor the metabolism of inositol phosphate derivatives in response to a mitogenic stimulus.
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Fibroblastos/citología , Péptido Hidrolasas/metabolismo , Calcio/metabolismo , División Celular/efectos de los fármacos , Membrana Celular/enzimología , Células Cultivadas , Medios de Cultivo , Fibroblastos/efectos de los fármacos , Fibroblastos/enzimología , Sustancias de Crecimiento/farmacología , Humanos , Fosfatos de Inositol/metabolismo , Cinética , alfa 1-Antitripsina/farmacologíaRESUMEN
Human alpha 1-proteinase inhibitor (alpha 1-antitrypsin) and bovine pancreatic trypsin inhibitor both inhibit cell-surface proteolytic activity in human fibroblasts. At the inhibitory concentrations, these proteins also inhibit cellular proliferation, as assayed by a microassay based on spectrophotometry of fixed and stained cells. Despite testing a wide range of proteinase inhibitor concentrations, no evidence was found for mitogenic or growth-stimulatory activity. ovomucoid was inactive against cell-surface proteinase activity and did not inhibit cellular proliferation.
Asunto(s)
División Celular/efectos de los fármacos , Inhibidores de Proteasas/farmacología , Ensayo de Inmunoadsorción Enzimática , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Humanos , Coloración y EtiquetadoRESUMEN
A report is made of serum zinc levels in 518 children with an age range of nine days to 17 years. There were significant differences in normal levels between fasting and non-fasting children, Europeans and Maoris, Europeans and Polynesians, and Indians and Polynesians. Lower levels were also found in those under the age of 13 but may be due to variation in serum proteins in these children who were all anaesthetised.