Paola Perez - Academia.edu (original) (raw)

Papers by Paola Perez

Annals of The Rheumatic Diseases, 2009

Objectives: In salivary glands from patients with Sjögren syndrome, overexpression of laminins 1 ... more Objectives: In salivary glands from patients with Sjögren syndrome, overexpression of laminins 1 and 5 and disorganisation of the acinar basal lamina have been reported. Laminin 5 mediates association of the basal lamina with epithelial cells by forming adhesion complexes upon interaction with a6b4 integrin. In the present work, mRNA and protein levels of a6b4 integrin were determined and its localisation in salivary glands evaluated in patients with Sjögren syndrome. Methods: Salivary glands of 12 patients with Sjögren syndrome and 8 controls were studied. The mRNA and protein levels of a6b4 were determined by semiquantitative reverse transcriptase (RT)-PCR and western blot analysis, respectively. The subcellular localisation of a6b4 and laminin were evaluated by confocal microscopy. Results: In patients, no significant differences in a6 and b4 mRNA levels were detected. However, b4 integrin protein levels were significantly lower, whereas, changes in a6, were highly variable. In controls, a6b4 was detected in the basolateral and basal surface of serous and mucous acini, respectively. In patients, alterations in a6b4 distribution were particularly dramatic for acini with strong basal lamina disorganisation. a6b4 was also detected in the cytoplasm and lateral plasma membrane in serous and mucous acini. Conclusion: Mild alterations in the basal lamina correlated with lateral redistribution of a6b4 integrin and the formation of new cell-cell adhesions that help maintain acinar organisation and promote cell survival. Conversely, in cases with severe basal lamina alterations, lateral a6b4 redistribution was no longer sufficient to maintain acinar cell survival. Thus, maintenance of equilibrium between cell-cell and cell-basal lamina attachment is required to sustain gland cell survival.

Annals of The Rheumatic Diseases, 2009

Objectives: In salivary glands from patients with Sjögren syndrome, overexpression of laminins 1 ... more Objectives: In salivary glands from patients with Sjögren syndrome, overexpression of laminins 1 and 5 and disorganisation of the acinar basal lamina have been reported. Laminin 5 mediates association of the basal lamina with epithelial cells by forming adhesion complexes upon interaction with a6b4 integrin. In the present work, mRNA and protein levels of a6b4 integrin were determined and its localisation in salivary glands evaluated in patients with Sjögren syndrome. Methods: Salivary glands of 12 patients with Sjögren syndrome and 8 controls were studied. The mRNA and protein levels of a6b4 were determined by semiquantitative reverse transcriptase (RT)-PCR and western blot analysis, respectively. The subcellular localisation of a6b4 and laminin were evaluated by confocal microscopy. Results: In patients, no significant differences in a6 and b4 mRNA levels were detected. However, b4 integrin protein levels were significantly lower, whereas, changes in a6, were highly variable. In controls, a6b4 was detected in the basolateral and basal surface of serous and mucous acini, respectively. In patients, alterations in a6b4 distribution were particularly dramatic for acini with strong basal lamina disorganisation. a6b4 was also detected in the cytoplasm and lateral plasma membrane in serous and mucous acini. Conclusion: Mild alterations in the basal lamina correlated with lateral redistribution of a6b4 integrin and the formation of new cell-cell adhesions that help maintain acinar organisation and promote cell survival. Conversely, in cases with severe basal lamina alterations, lateral a6b4 redistribution was no longer sufficient to maintain acinar cell survival. Thus, maintenance of equilibrium between cell-cell and cell-basal lamina attachment is required to sustain gland cell survival.

Radiotherapy and Oncology, 1996

Radiotherapy and Oncology, 1996

Rheumatology, 2010

Objectives. To analyse whether the alterations in the structure and organization of microvilli in... more Objectives. To analyse whether the alterations in the structure and organization of microvilli in salivary acinar cells from SS patients are linked to changes in the expression and/or cellular localization of ezrin.

Rheumatology, 2010

Objectives. To analyse whether the alterations in the structure and organization of microvilli in... more Objectives. To analyse whether the alterations in the structure and organization of microvilli in salivary acinar cells from SS patients are linked to changes in the expression and/or cellular localization of ezrin.

Biochemical and Biophysical Research Communications, 2010

Polarized salivary epithelial cells can sort secretory proteins towards either the basolateral or... more Polarized salivary epithelial cells can sort secretory proteins towards either the basolateral or apical pole. Transgenic human parathyroid hormone (hPTH) exclusively sorts apically in rat submandibular glands. To help understand this specific process we modified the hPTH cDNA sequence and delivered the cDNAs to glands in vivo using adenoviral (Ad) vectors. The Ad vectors encoded: 1) the native form of hPTH (Ad.pre-pro-hPTH1-84), 2) the native sequence, but with the pro segment deleted (Ad.pre-hPTH1-84), and 3) a sequence containing the pre segment followed by the first 34 amino acids of hPTH (Ad.pre-hPTH1-34). hPTH production and sorting were studied after two days. All constructs were effectively transcribed in targeted glands. However, the pre-hPTH1-84 modification led to reduced hPTH secretion and production, while no immunoreactive hPTH resulted from pre-hPTH1-34 cDNA infusion. The pre-hPTH1-84 modification had no effect on apical sorting. These in vivo results show that the signal responsible for hPTH's apical sorting does not reside in the pro segment and that deleting both the pro segment and the carboxyl-terminal region severely impairs post-translational processing of hPTH.

Human Gene Therapy, 2008

Gene transfer to salivary glands leads to abundant secretion of transgenic protein into either sa... more Gene transfer to salivary glands leads to abundant secretion of transgenic protein into either saliva or the bloodstream. This indicates significant clinical potential, depending on the route of sorting. The aim of this study was to probe the sorting characteristics of human parathyroid hormone (hPTH) in two animal models for salivary gland gene transfer. PTH is a key hormone regulating calcium levels in the blood. A recombinant serotype 5 adenoviral vector carrying the hPTH cDNA was administered to the submandibular glands of mice and rats. Two days after delivery, high levels of hPTH were found in the serum of mice, leading to elevated serum calcium levels. Only low amounts of hPTH were found in the saliva. Two days after vector infusion into rats, a massive secretion of hPTH was measured in saliva, with little secretion into serum. Confocal microscopy showed hPTH in the glands, localized basolaterally in mice and apically in rats. Submandibular gland transduction was effective and the produced hPTH was biologically active in vivo. Whereas hPTH sorted toward the basolateral side in mice, in rats hPTH was secreted mainly at the apical side. These results indicate that the interaction between hPTH and the cell sorting machinery is different between mouse and rat salivary glands. Detailed studies in these two species should result in a better understanding of cellular control of transgenic secretory protein sorting in this tissue.

International Journal of Biochemistry & Cell Biology, 2010

Salivary glands are classical exocrine glands whose external secretions result in the production ... more Salivary glands are classical exocrine glands whose external secretions result in the production of saliva. However, in addition to the secretion of exocrine proteins, salivary epithelial cells are also capable of secreting proteins internally, into the bloodstream. This brief review examines the potential for using salivary epithelial cells as a target site for in situ gene transfer, with an ultimate goal of producing therapeutic proteins for treating both systemic and upper gastrointestinal tract disorders. The review discusses the protein secretory pathways reported to be present in salivary epithelial cells, the viral gene transfer vectors shown useful for transducing these cells, model transgenic secretory proteins examined, and some clinical conditions that might benefit from such salivary gland gene transfer.

Biochemical and Biophysical Research Communications, 2010

Polarized salivary epithelial cells can sort secretory proteins towards either the basolateral or... more Polarized salivary epithelial cells can sort secretory proteins towards either the basolateral or apical pole. Transgenic human parathyroid hormone (hPTH) exclusively sorts apically in rat submandibular glands. To help understand this specific process we modified the hPTH cDNA sequence and delivered the cDNAs to glands in vivo using adenoviral (Ad) vectors. The Ad vectors encoded: 1) the native form of hPTH (Ad.pre-pro-hPTH1-84), 2) the native sequence, but with the pro segment deleted (Ad.pre-hPTH1-84), and 3) a sequence containing the pre segment followed by the first 34 amino acids of hPTH (Ad.pre-hPTH1-34). hPTH production and sorting were studied after two days. All constructs were effectively transcribed in targeted glands. However, the pre-hPTH1-84 modification led to reduced hPTH secretion and production, while no immunoreactive hPTH resulted from pre-hPTH1-34 cDNA infusion. The pre-hPTH1-84 modification had no effect on apical sorting. These in vivo results show that the signal responsible for hPTH's apical sorting does not reside in the pro segment and that deleting both the pro segment and the carboxyl-terminal region severely impairs post-translational processing of hPTH.

Human Gene Therapy, 2008

Gene transfer to salivary glands leads to abundant secretion of transgenic protein into either sa... more Gene transfer to salivary glands leads to abundant secretion of transgenic protein into either saliva or the bloodstream. This indicates significant clinical potential, depending on the route of sorting. The aim of this study was to probe the sorting characteristics of human parathyroid hormone (hPTH) in two animal models for salivary gland gene transfer. PTH is a key hormone regulating calcium levels in the blood. A recombinant serotype 5 adenoviral vector carrying the hPTH cDNA was administered to the submandibular glands of mice and rats. Two days after delivery, high levels of hPTH were found in the serum of mice, leading to elevated serum calcium levels. Only low amounts of hPTH were found in the saliva. Two days after vector infusion into rats, a massive secretion of hPTH was measured in saliva, with little secretion into serum. Confocal microscopy showed hPTH in the glands, localized basolaterally in mice and apically in rats. Submandibular gland transduction was effective and the produced hPTH was biologically active in vivo. Whereas hPTH sorted toward the basolateral side in mice, in rats hPTH was secreted mainly at the apical side. These results indicate that the interaction between hPTH and the cell sorting machinery is different between mouse and rat salivary glands. Detailed studies in these two species should result in a better understanding of cellular control of transgenic secretory protein sorting in this tissue.

International Journal of Biochemistry & Cell Biology, 2010

Salivary glands are classical exocrine glands whose external secretions result in the production ... more Salivary glands are classical exocrine glands whose external secretions result in the production of saliva. However, in addition to the secretion of exocrine proteins, salivary epithelial cells are also capable of secreting proteins internally, into the bloodstream. This brief review examines the potential for using salivary epithelial cells as a target site for in situ gene transfer, with an ultimate goal of producing therapeutic proteins for treating both systemic and upper gastrointestinal tract disorders. The review discusses the protein secretory pathways reported to be present in salivary epithelial cells, the viral gene transfer vectors shown useful for transducing these cells, model transgenic secretory proteins examined, and some clinical conditions that might benefit from such salivary gland gene transfer.

Annals of The Rheumatic Diseases, 2009

Objectives: In salivary glands from patients with Sjögren syndrome, overexpression of laminins 1 ... more Objectives: In salivary glands from patients with Sjögren syndrome, overexpression of laminins 1 and 5 and disorganisation of the acinar basal lamina have been reported. Laminin 5 mediates association of the basal lamina with epithelial cells by forming adhesion complexes upon interaction with a6b4 integrin. In the present work, mRNA and protein levels of a6b4 integrin were determined and its localisation in salivary glands evaluated in patients with Sjögren syndrome. Methods: Salivary glands of 12 patients with Sjögren syndrome and 8 controls were studied. The mRNA and protein levels of a6b4 were determined by semiquantitative reverse transcriptase (RT)-PCR and western blot analysis, respectively. The subcellular localisation of a6b4 and laminin were evaluated by confocal microscopy. Results: In patients, no significant differences in a6 and b4 mRNA levels were detected. However, b4 integrin protein levels were significantly lower, whereas, changes in a6, were highly variable. In controls, a6b4 was detected in the basolateral and basal surface of serous and mucous acini, respectively. In patients, alterations in a6b4 distribution were particularly dramatic for acini with strong basal lamina disorganisation. a6b4 was also detected in the cytoplasm and lateral plasma membrane in serous and mucous acini. Conclusion: Mild alterations in the basal lamina correlated with lateral redistribution of a6b4 integrin and the formation of new cell-cell adhesions that help maintain acinar organisation and promote cell survival. Conversely, in cases with severe basal lamina alterations, lateral a6b4 redistribution was no longer sufficient to maintain acinar cell survival. Thus, maintenance of equilibrium between cell-cell and cell-basal lamina attachment is required to sustain gland cell survival.

Annals of The Rheumatic Diseases, 2009

Objectives: In salivary glands from patients with Sjögren syndrome, overexpression of laminins 1 ... more Objectives: In salivary glands from patients with Sjögren syndrome, overexpression of laminins 1 and 5 and disorganisation of the acinar basal lamina have been reported. Laminin 5 mediates association of the basal lamina with epithelial cells by forming adhesion complexes upon interaction with a6b4 integrin. In the present work, mRNA and protein levels of a6b4 integrin were determined and its localisation in salivary glands evaluated in patients with Sjögren syndrome. Methods: Salivary glands of 12 patients with Sjögren syndrome and 8 controls were studied. The mRNA and protein levels of a6b4 were determined by semiquantitative reverse transcriptase (RT)-PCR and western blot analysis, respectively. The subcellular localisation of a6b4 and laminin were evaluated by confocal microscopy. Results: In patients, no significant differences in a6 and b4 mRNA levels were detected. However, b4 integrin protein levels were significantly lower, whereas, changes in a6, were highly variable. In controls, a6b4 was detected in the basolateral and basal surface of serous and mucous acini, respectively. In patients, alterations in a6b4 distribution were particularly dramatic for acini with strong basal lamina disorganisation. a6b4 was also detected in the cytoplasm and lateral plasma membrane in serous and mucous acini. Conclusion: Mild alterations in the basal lamina correlated with lateral redistribution of a6b4 integrin and the formation of new cell-cell adhesions that help maintain acinar organisation and promote cell survival. Conversely, in cases with severe basal lamina alterations, lateral a6b4 redistribution was no longer sufficient to maintain acinar cell survival. Thus, maintenance of equilibrium between cell-cell and cell-basal lamina attachment is required to sustain gland cell survival.

Radiotherapy and Oncology, 1996

Radiotherapy and Oncology, 1996

Rheumatology, 2010

Objectives. To analyse whether the alterations in the structure and organization of microvilli in... more Objectives. To analyse whether the alterations in the structure and organization of microvilli in salivary acinar cells from SS patients are linked to changes in the expression and/or cellular localization of ezrin.

Rheumatology, 2010

Objectives. To analyse whether the alterations in the structure and organization of microvilli in... more Objectives. To analyse whether the alterations in the structure and organization of microvilli in salivary acinar cells from SS patients are linked to changes in the expression and/or cellular localization of ezrin.

Biochemical and Biophysical Research Communications, 2010

Polarized salivary epithelial cells can sort secretory proteins towards either the basolateral or... more Polarized salivary epithelial cells can sort secretory proteins towards either the basolateral or apical pole. Transgenic human parathyroid hormone (hPTH) exclusively sorts apically in rat submandibular glands. To help understand this specific process we modified the hPTH cDNA sequence and delivered the cDNAs to glands in vivo using adenoviral (Ad) vectors. The Ad vectors encoded: 1) the native form of hPTH (Ad.pre-pro-hPTH1-84), 2) the native sequence, but with the pro segment deleted (Ad.pre-hPTH1-84), and 3) a sequence containing the pre segment followed by the first 34 amino acids of hPTH (Ad.pre-hPTH1-34). hPTH production and sorting were studied after two days. All constructs were effectively transcribed in targeted glands. However, the pre-hPTH1-84 modification led to reduced hPTH secretion and production, while no immunoreactive hPTH resulted from pre-hPTH1-34 cDNA infusion. The pre-hPTH1-84 modification had no effect on apical sorting. These in vivo results show that the signal responsible for hPTH's apical sorting does not reside in the pro segment and that deleting both the pro segment and the carboxyl-terminal region severely impairs post-translational processing of hPTH.

Human Gene Therapy, 2008

Gene transfer to salivary glands leads to abundant secretion of transgenic protein into either sa... more Gene transfer to salivary glands leads to abundant secretion of transgenic protein into either saliva or the bloodstream. This indicates significant clinical potential, depending on the route of sorting. The aim of this study was to probe the sorting characteristics of human parathyroid hormone (hPTH) in two animal models for salivary gland gene transfer. PTH is a key hormone regulating calcium levels in the blood. A recombinant serotype 5 adenoviral vector carrying the hPTH cDNA was administered to the submandibular glands of mice and rats. Two days after delivery, high levels of hPTH were found in the serum of mice, leading to elevated serum calcium levels. Only low amounts of hPTH were found in the saliva. Two days after vector infusion into rats, a massive secretion of hPTH was measured in saliva, with little secretion into serum. Confocal microscopy showed hPTH in the glands, localized basolaterally in mice and apically in rats. Submandibular gland transduction was effective and the produced hPTH was biologically active in vivo. Whereas hPTH sorted toward the basolateral side in mice, in rats hPTH was secreted mainly at the apical side. These results indicate that the interaction between hPTH and the cell sorting machinery is different between mouse and rat salivary glands. Detailed studies in these two species should result in a better understanding of cellular control of transgenic secretory protein sorting in this tissue.

International Journal of Biochemistry & Cell Biology, 2010

Salivary glands are classical exocrine glands whose external secretions result in the production ... more Salivary glands are classical exocrine glands whose external secretions result in the production of saliva. However, in addition to the secretion of exocrine proteins, salivary epithelial cells are also capable of secreting proteins internally, into the bloodstream. This brief review examines the potential for using salivary epithelial cells as a target site for in situ gene transfer, with an ultimate goal of producing therapeutic proteins for treating both systemic and upper gastrointestinal tract disorders. The review discusses the protein secretory pathways reported to be present in salivary epithelial cells, the viral gene transfer vectors shown useful for transducing these cells, model transgenic secretory proteins examined, and some clinical conditions that might benefit from such salivary gland gene transfer.

Biochemical and Biophysical Research Communications, 2010

Polarized salivary epithelial cells can sort secretory proteins towards either the basolateral or... more Polarized salivary epithelial cells can sort secretory proteins towards either the basolateral or apical pole. Transgenic human parathyroid hormone (hPTH) exclusively sorts apically in rat submandibular glands. To help understand this specific process we modified the hPTH cDNA sequence and delivered the cDNAs to glands in vivo using adenoviral (Ad) vectors. The Ad vectors encoded: 1) the native form of hPTH (Ad.pre-pro-hPTH1-84), 2) the native sequence, but with the pro segment deleted (Ad.pre-hPTH1-84), and 3) a sequence containing the pre segment followed by the first 34 amino acids of hPTH (Ad.pre-hPTH1-34). hPTH production and sorting were studied after two days. All constructs were effectively transcribed in targeted glands. However, the pre-hPTH1-84 modification led to reduced hPTH secretion and production, while no immunoreactive hPTH resulted from pre-hPTH1-34 cDNA infusion. The pre-hPTH1-84 modification had no effect on apical sorting. These in vivo results show that the signal responsible for hPTH's apical sorting does not reside in the pro segment and that deleting both the pro segment and the carboxyl-terminal region severely impairs post-translational processing of hPTH.

Human Gene Therapy, 2008

Gene transfer to salivary glands leads to abundant secretion of transgenic protein into either sa... more Gene transfer to salivary glands leads to abundant secretion of transgenic protein into either saliva or the bloodstream. This indicates significant clinical potential, depending on the route of sorting. The aim of this study was to probe the sorting characteristics of human parathyroid hormone (hPTH) in two animal models for salivary gland gene transfer. PTH is a key hormone regulating calcium levels in the blood. A recombinant serotype 5 adenoviral vector carrying the hPTH cDNA was administered to the submandibular glands of mice and rats. Two days after delivery, high levels of hPTH were found in the serum of mice, leading to elevated serum calcium levels. Only low amounts of hPTH were found in the saliva. Two days after vector infusion into rats, a massive secretion of hPTH was measured in saliva, with little secretion into serum. Confocal microscopy showed hPTH in the glands, localized basolaterally in mice and apically in rats. Submandibular gland transduction was effective and the produced hPTH was biologically active in vivo. Whereas hPTH sorted toward the basolateral side in mice, in rats hPTH was secreted mainly at the apical side. These results indicate that the interaction between hPTH and the cell sorting machinery is different between mouse and rat salivary glands. Detailed studies in these two species should result in a better understanding of cellular control of transgenic secretory protein sorting in this tissue.

International Journal of Biochemistry & Cell Biology, 2010

Salivary glands are classical exocrine glands whose external secretions result in the production ... more Salivary glands are classical exocrine glands whose external secretions result in the production of saliva. However, in addition to the secretion of exocrine proteins, salivary epithelial cells are also capable of secreting proteins internally, into the bloodstream. This brief review examines the potential for using salivary epithelial cells as a target site for in situ gene transfer, with an ultimate goal of producing therapeutic proteins for treating both systemic and upper gastrointestinal tract disorders. The review discusses the protein secretory pathways reported to be present in salivary epithelial cells, the viral gene transfer vectors shown useful for transducing these cells, model transgenic secretory proteins examined, and some clinical conditions that might benefit from such salivary gland gene transfer.