AACR Annual Meeting-- Apr 14-18, 2007; Los Angeles, CA 3029 The selectins are a family of interce... more AACR Annual Meeting-- Apr 14-18, 2007; Los Angeles, CA 3029 The selectins are a family of intercellular adhesion molecules with three members: E-, P- and L-selectin. E-selectin (ELAM-1, CD62E) is a type Ia transmembrane protein containing lectin-like and endothelial growth factor-like domains, followed by short cysteine-rich repeats. Absent in resting epithelium, E-selectin is rapidly expressed in the vascular lining during inflammation. Indeed, activation of endothelial cells by proinflammatory cytokines results in the cell surface appearance of E-selectin, facilitating leukocyte rolling and extravasation through activated endothelium. During a screening of gene expression in several human colon cancer cell lines we surprisingly identified E-selectin, as a molecule that was modulated in response to TNF-α. The up-regulation of E-selectin was also confirmed at the protein level with a mAb against human E-selectin. In order to better understand the role of E-selectin in the expression of tumor phenotype, we transfected T84 human colon cancer cells with the full length E-selectin cDNA and cDNA coding for the E-selectin shedded domain. We selected clones with a variable expression of the proteins and tested them for their ability to form aggregates, migrate, establish colonies in soft agar as well as to adhere to extracellular matrix substrates. The results we obtained strongly support the hypothesis that E-selectin plays an active role in metastatic cascade not only determining the extravasation site for tumor cells but also modulating growth rate and survival. Acknowledgments This work was supported by Italian Association for Cancer Research (AIRC)
Fabry disease (FD) is an X-linked progressive multisystem disease due to mutations in the gene en... more Fabry disease (FD) is an X-linked progressive multisystem disease due to mutations in the gene encoding the lysosomal enzyme α-galactosidase A (α-GalA). The deficiency in α-GalA activity leads to an intra-lysosomal accumulation of neutral glycosphingolipids, mainly globotriaosylceramide (Gb3), in various organs and systems. Enzyme replacement therapy is available and alternative therapeutic approaches are being explored. No diagnostic test, other than sequencing of the α-galactosidase A gene, is available, no biomarker has been proven useful to screen for and predict the disease, and underlying mechanisms are still elusive. The aim of this study is to identify FD specific biomarkers and to better understand the pathophysiological changes that occur over time in FD. We compared peripheral blood mononuclear cells (PBMC) from FD patients (n = 8) with control PBMC from healthy individuals (n = 6), by two-dimensional electrophoresis (2DE) and the detected differentially expressed proteins were then subjected to matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS). In FD patients we identified, among the down-regulated proteins, Calnexin, Rho GDP-dissociation inhibitor 2, Rho GDP-dissociation inhibitor 1, Chloride intracellular channel protein 1; on the other hand γ-enolase, 14-3-3 protein theta, 14-3-3 protein zeta/delta, and galectin-1 were identified as up-regulated proteins. Calnexin and Rho GDP-dissociation inhibitor-1,2 are related to protein folding, signal transduction and cell proliferation. This is the first time that γ-enolase and galectin-1 are described to be up-regulated in Fabry patients. Levels of γ-enolase increase dramatically in cardiovascular accidents and cerebral trauma, whereas galectins are regulators of acute and chronic inflammation. These findings may improve our understanding of the molecular mechanisms underlying the pathology and provide new insight and knowledge for future studies in this field.
Pompe disease (PD), also defined as acid maltase deficiency, is a rare autosomal recessive diseas... more Pompe disease (PD), also defined as acid maltase deficiency, is a rare autosomal recessive disease that causes glycogen accumulation due to a deficiency of the lysosomal enzyme acid α-glucosidase. An excessive amount of undisposed glycogen causes progressive muscle weakness throughout the body. It particularly affects skeletal muscles and the nervous system, especially in the late-onset phase. Here, we present a clinical case of late-onset PD (LOPD) with normal CK (creatinine kinase) values treated after a misdiagnosis of demyelinating motor polyneuropathy and chronic inflammatory neuropathy. The suspicion of possible fibromyalgia induced the patient to seek a rheumatology consultation, and the investigations performed led to the diagnosis of PD. The patient was investigated for genetic and enzymatic studies. PD was diagnosed using the α-glucosidase assay on DBS. In LOPD, clinical manifestations, such as muscle weakness, exercise intolerance, myalgia, or even high hyperCKemia, often...
AACR Annual Meeting-- Apr 14-18, 2007; Los Angeles, CA 3029 The selectins are a family of interce... more AACR Annual Meeting-- Apr 14-18, 2007; Los Angeles, CA 3029 The selectins are a family of intercellular adhesion molecules with three members: E-, P- and L-selectin. E-selectin (ELAM-1, CD62E) is a type Ia transmembrane protein containing lectin-like and endothelial growth factor-like domains, followed by short cysteine-rich repeats. Absent in resting epithelium, E-selectin is rapidly expressed in the vascular lining during inflammation. Indeed, activation of endothelial cells by proinflammatory cytokines results in the cell surface appearance of E-selectin, facilitating leukocyte rolling and extravasation through activated endothelium. During a screening of gene expression in several human colon cancer cell lines we surprisingly identified E-selectin, as a molecule that was modulated in response to TNF-α. The up-regulation of E-selectin was also confirmed at the protein level with a mAb against human E-selectin. In order to better understand the role of E-selectin in the expression of tumor phenotype, we transfected T84 human colon cancer cells with the full length E-selectin cDNA and cDNA coding for the E-selectin shedded domain. We selected clones with a variable expression of the proteins and tested them for their ability to form aggregates, migrate, establish colonies in soft agar as well as to adhere to extracellular matrix substrates. The results we obtained strongly support the hypothesis that E-selectin plays an active role in metastatic cascade not only determining the extravasation site for tumor cells but also modulating growth rate and survival. Acknowledgments This work was supported by Italian Association for Cancer Research (AIRC)
Fabry disease (FD) is an X-linked progressive multisystem disease due to mutations in the gene en... more Fabry disease (FD) is an X-linked progressive multisystem disease due to mutations in the gene encoding the lysosomal enzyme α-galactosidase A (α-GalA). The deficiency in α-GalA activity leads to an intra-lysosomal accumulation of neutral glycosphingolipids, mainly globotriaosylceramide (Gb3), in various organs and systems. Enzyme replacement therapy is available and alternative therapeutic approaches are being explored. No diagnostic test, other than sequencing of the α-galactosidase A gene, is available, no biomarker has been proven useful to screen for and predict the disease, and underlying mechanisms are still elusive. The aim of this study is to identify FD specific biomarkers and to better understand the pathophysiological changes that occur over time in FD. We compared peripheral blood mononuclear cells (PBMC) from FD patients (n = 8) with control PBMC from healthy individuals (n = 6), by two-dimensional electrophoresis (2DE) and the detected differentially expressed proteins were then subjected to matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS). In FD patients we identified, among the down-regulated proteins, Calnexin, Rho GDP-dissociation inhibitor 2, Rho GDP-dissociation inhibitor 1, Chloride intracellular channel protein 1; on the other hand γ-enolase, 14-3-3 protein theta, 14-3-3 protein zeta/delta, and galectin-1 were identified as up-regulated proteins. Calnexin and Rho GDP-dissociation inhibitor-1,2 are related to protein folding, signal transduction and cell proliferation. This is the first time that γ-enolase and galectin-1 are described to be up-regulated in Fabry patients. Levels of γ-enolase increase dramatically in cardiovascular accidents and cerebral trauma, whereas galectins are regulators of acute and chronic inflammation. These findings may improve our understanding of the molecular mechanisms underlying the pathology and provide new insight and knowledge for future studies in this field.
Pompe disease (PD), also defined as acid maltase deficiency, is a rare autosomal recessive diseas... more Pompe disease (PD), also defined as acid maltase deficiency, is a rare autosomal recessive disease that causes glycogen accumulation due to a deficiency of the lysosomal enzyme acid α-glucosidase. An excessive amount of undisposed glycogen causes progressive muscle weakness throughout the body. It particularly affects skeletal muscles and the nervous system, especially in the late-onset phase. Here, we present a clinical case of late-onset PD (LOPD) with normal CK (creatinine kinase) values treated after a misdiagnosis of demyelinating motor polyneuropathy and chronic inflammatory neuropathy. The suspicion of possible fibromyalgia induced the patient to seek a rheumatology consultation, and the investigations performed led to the diagnosis of PD. The patient was investigated for genetic and enzymatic studies. PD was diagnosed using the α-glucosidase assay on DBS. In LOPD, clinical manifestations, such as muscle weakness, exercise intolerance, myalgia, or even high hyperCKemia, often...
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