RESUMEN
AQP5 plays an important role in the salivary gland function. The mRNA and protein for aquaporin 5 (AQP5) are expressed in the acini from embryonic days E13-16 and E17-18, respectively and for entire postnatal days. Ligation-reopening of main excretory duct induces changes in the AQP5 level which would give an insight for mechanism of regeneration/self-duplication of acinar cells. The AQP5 level in the submandibular gland (SMG) decreases by chorda tympani denervation (CTD) via activation autophagosome, suggesting that its level in the SMG under normal condition is maintained by parasympathetic nerve. Isoproterenol (IPR), a ß-adrenergic agonist, raised the levels of membrane AQP5 protein and its mRNA in the parotid gland (PG), suggesting coupling of the AQP5 dynamic and amylase secretion-restoration cycle. In the PG, lipopolysaccharide (LPS) is shown to activate mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) signalings and potentially downregulate AQP5 expression via cross coupling of activator protein-1 (AP-1) and NF-κB. In most species, Ser-156 and Thr-259 of AQP5 are experimentally phosphorylated, which is enhanced by cAMP analogues and forskolin. cAMP-dependent phosphorylation of AQP5 does not seem to be markedly involved in regulation of its intracellular trafficking but seems to play a role in its constitutive expression and lateral diffusion in the cell membrane. Additionally, Ser-156 phosphorylation may be important for cancer development.
Asunto(s)
Acuaporina 5/metabolismo , Glándulas Salivales/fisiología , Animales , Acuaporina 5/análisis , Acuaporina 5/genética , Regulación de la Expresión Génica , Humanos , Fosforilación , Procesamiento Proteico-Postraduccional , Enfermedades de las Glándulas Salivales/genética , Enfermedades de las Glándulas Salivales/metabolismo , Enfermedades de las Glándulas Salivales/fisiopatología , Glándulas Salivales/crecimiento & desarrollo , Glándulas Salivales/fisiopatología , UbiquitinaciónRESUMEN
Aplasia/agenesis of lacrimal and salivary glands is a rare congenital defect that has been associated with disturbances in fibroblast growth factor 10 (FGF10). It can present with symptoms of congenital lacrimal obstruction, dry eye, and dry mouth. We report the ophthalmological and genetic study of a 19-year-old woman and her relatives suffering from this syndrome. A new probably pathogenic variant is described in the FGF10 gene.
Asunto(s)
Anomalías del Ojo/genética , Factor 10 de Crecimiento de Fibroblastos/genética , Enfermedades del Aparato Lagrimal/genética , Aparato Lagrimal/anomalías , Enfermedades de las Glándulas Salivales/genética , Glándulas Salivales/anomalías , Síndromes de Ojo Seco/diagnóstico , Anomalías del Ojo/diagnóstico por imagen , Femenino , Humanos , Aparato Lagrimal/diagnóstico por imagen , Enfermedades del Aparato Lagrimal/diagnóstico por imagen , Obstrucción del Conducto Lagrimal/diagnóstico , Imagen por Resonancia Magnética , Linaje , Enfermedades de las Glándulas Salivales/diagnóstico por imagen , Glándulas Salivales/diagnóstico por imagen , Xerostomía/diagnóstico , Adulto JovenRESUMEN
For many years, our research group worked to develop gene transfer approaches for salivary gland disorders that lacked effective conventional therapy. The purpose of this chapter is to describe and update key methods used in this process. As described in our original chapter from the 2010 volume, we focus on one clinical condition, irradiation-induced salivary hypofunction, and address the choice of transgene and vector to be used, the construction of recombinant viral vectors, how vector delivery is accomplished, and methods for assessing vector function in vitro and in an appropriate animal model.
Asunto(s)
Terapia Genética , Enfermedades de las Glándulas Salivales/genética , Enfermedades de las Glándulas Salivales/terapia , Adenovirus Humanos/genética , Animales , Línea Celular , Dependovirus , Modelos Animales de Enfermedad , Orden Génico , Técnicas de Transferencia de Gen , Terapia Genética/métodos , Vectores Genéticos/administración & dosificación , Vectores Genéticos/genética , Vectores Genéticos/aislamiento & purificación , Humanos , Parvovirinae/genética , Glándulas Salivales/metabolismo , Transducción Genética , TransgenesRESUMEN
In the pupal stage, the fly body undergoes extensive metamorphic remodeling, in which programmed cell death plays a critical role. We studied two of the constituent processes in this remodeling, salivary gland degeneration and breakdown of the eclosion muscle, which are triggered by an increase and a decrease in the circulating steroid hormone ecdysone at the start and end of metamorphosis, respectively. We found that knockdown of zeste (z), a gene encoding a sequence-specific DNA-binding protein implicated in transvection, in salivary gland cells advances the initiation of their degeneration, whereas z knockdown in neurons delays muscle breakdown. We further showed that knockdown of an ecdysone-inducible gene, E74, retards salivary gland degeneration with little effect on eclosion muscle breakdown. We propose that Z tunes the sensitivity of ecdysone targets to this hormone in order to ensure a high safety margin so that the cell death program will be activated when the ecdysone titer is at a sufficiently high level that is reached only at a defined stage during metamorphosis.
Asunto(s)
Proteínas de Unión al ADN/metabolismo , Proteínas de Drosophila/metabolismo , Ecdisona/metabolismo , Regulación del Desarrollo de la Expresión Génica/genética , Enfermedades de las Glándulas Salivales/genética , Glándulas Salivales/patología , Factores de Edad , Animales , Animales Modificados Genéticamente , Proteínas de Unión al ADN/genética , Drosophila , Proteínas de Drosophila/genética , Músculos , Enfermedades Musculares/genética , Pupa , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismoRESUMEN
OBJECTIVES: Primary SS is an autoimmune disease characterized by chronic lymphocytic inflammation and ectopic germinal centre (GC) formation within salivary glands. Fractalkine (CX3CL1), associated with the pathogenesis of RA, is the sole member of the CX3C chemokine (CK) family and acts as an adhesion and chemotactic molecule. The objectives of this work are to determine to what extent CX3CL1 and its receptor CX3CR1 expression might be altered in salivary glands obtained from patients and to establish whether these CKs might be involved in SS ectopic lymphoneogenesis. METHODS: We assessed the presence of CX3CL1 protein in sera by ELISA in 21 patients with primary SS, 11 patients with Sicca syndrome (Sicca), 20 RA patients and 10 blood donors. Histological evaluation was performed on sequential sections of salivary gland tissue. Using TaqMan RT-PCR we studied CX3CL1 and CX3CR1 mRNA expression in salivary gland tissues from a molecular point of view. RESULTS: Increased serum levels of CX3CL1 protein were observed in SS patients compared with controls (P < 0.0001) and in RA patients compared with controls (P < 0.0001), but no difference was found between Sicca patients and controls (P = 0.22). We identified histologically the cells expressing CX3CL1 and CX3CR1 in salivary glands of SS patients and we localized the molecule within tertiary lymphoid structures. Finally, the mRNA levels of the CK and its receptor were up-regulated in SS salivary glands. CONCLUSION: We believe that our findings point to the need for future studies on CX3CL1 and CX3CR1 proteins as contributors to the formation of ectopic GCs and possibly as a new tool in the evaluation and diagnosis of SS.
Asunto(s)
Quimiocina CX3CL1/inmunología , Coristoma/inmunología , Tejido Linfoide/inmunología , ARN Mensajero/análisis , Receptores de Quimiocina/inmunología , Enfermedades de las Glándulas Salivales/inmunología , Síndrome de Sjögren/inmunología , Adulto , Anciano , Artritis Reumatoide/genética , Artritis Reumatoide/inmunología , Artritis Reumatoide/patología , Receptor 1 de Quimiocinas CX3C , Estudios de Casos y Controles , Quimiocina CX3CL1/genética , Coristoma/genética , Coristoma/patología , Femenino , Centro Germinal/inmunología , Humanos , Tejido Linfoide/patología , Masculino , Persona de Mediana Edad , Receptores de Quimiocina/genética , Enfermedades de las Glándulas Salivales/genética , Enfermedades de las Glándulas Salivales/patología , Glándulas Salivales Menores/inmunología , Glándulas Salivales Menores/patología , Síndrome de Sjögren/genética , Síndrome de Sjögren/patología , Adulto JovenRESUMEN
BACKGROUND & AIMS: Excessive Ca2+ influx mediates many cytotoxic processes, including those associated with autoimmune inflammatory diseases such as acute pancreatitis and Sjögren syndrome. Transient receptor potential (canonical) channel (TRPC) 3 is a major Ca2+ influx channel in pancreatic and salivary gland cells. We investigated whether genetic or pharmacologic inhibition of TRPC3 protects pancreas and salivary glands from Ca2+-dependent damage. METHODS: We developed a Ca2+-dependent model of cell damage for salivary gland acini. Acute pancreatitis was induced by injection of cerulein into wild-type and Trpc3-/- mice. Mice were also given the Trpc3-selective inhibitor pyrazole 3 (Pyr3). RESULTS: Salivary glands and pancreas of Trpc3-/- mice were protected from Ca2+-mediated cell toxicity. Analysis of Ca2+ signaling in wild-type and Trpc3-/- acini showed that Pyr3 is a highly specific inhibitor of Tprc3; it protected salivary glands and pancreas cells from Ca2+-mediated toxicity by inhibiting the Trpc3-mediated component of Ca2+ influx. CONCLUSIONS: TRPC3-mediated Ca2+ influx mediates damage to pancreas and salivary glands. Pharmacologic inhibition of TRPC3 with the highly selective TRPC3 inhibitor Pyr3 might be developed for treatment of patients with acute pancreatitis and Sjögren syndrome.
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Bloqueadores de los Canales de Calcio/farmacología , Señalización del Calcio/efectos de los fármacos , Células Epiteliales/efectos de los fármacos , Páncreas/efectos de los fármacos , Pancreatitis/tratamiento farmacológico , Pirazoles/farmacología , Enfermedades de las Glándulas Salivales/tratamiento farmacológico , Glándulas Salivales/efectos de los fármacos , Canales Catiónicos TRPC/antagonistas & inhibidores , Enfermedad Aguda , Animales , Ceruletida , Modelos Animales de Enfermedad , Células Epiteliales/metabolismo , Células Epiteliales/patología , Ratones , Ratones Noqueados , Páncreas/metabolismo , Páncreas/patología , Pancreatitis/inducido químicamente , Pancreatitis/genética , Pancreatitis/metabolismo , Pancreatitis/patología , Enfermedades de las Glándulas Salivales/genética , Enfermedades de las Glándulas Salivales/metabolismo , Enfermedades de las Glándulas Salivales/patología , Glándulas Salivales/metabolismo , Glándulas Salivales/patología , Índice de Severidad de la Enfermedad , Canales Catiónicos TRPC/deficiencia , Canales Catiónicos TRPC/genética , Factores de TiempoRESUMEN
Branching morphogenesis is a crucial developmental process in which vertebrate organs generate extensive epithelial surface area while retaining a compact size. In the vertebrate submandibular salivary gland, branching morphogenesis is crucial for the generation of the large surface area necessary to produce sufficient saliva. However, in many salivary gland diseases, saliva-producing acinar cells are destroyed, resulting in dry mouth and secondary health conditions. Systems-based approaches can provide insights into understanding salivary gland development, function, and disease. The traditional approach to understanding these processes is the identification of molecular signals using reductionist approaches; we review current progress with such methods in understanding salivary gland development. Taking a more global approach, multiple groups are currently profiling the transcriptome, the proteome, and other 'omes' in both developing mouse tissues and in human patient samples. Computational methods have been successful in deciphering large data sets, and mathematical models are starting to make predictions regarding the contribution of molecules to the physical processes of morphogenesis and cellular function. A challenge for the future will be to establish comprehensive, publicly accessible salivary gland databases spanning the full range of genes and proteins; plans are underway to provide these resources to researchers in centralized repositories. The greatest challenge for the future will be to develop realistic models that integrate multiple types of data to both describe and predict embryonic development and disease pathogenesis.
Asunto(s)
Enfermedades de las Glándulas Salivales/metabolismo , Glándulas Salivales/crecimiento & desarrollo , Animales , Biología Computacional , Perfilación de la Expresión Génica , Humanos , Redes y Vías Metabólicas , Ratones , MicroARNs/metabolismo , Modelos Teóricos , Morfogénesis , Proteoma , Enfermedades de las Glándulas Salivales/genética , Neoplasias de las Glándulas Salivales/genética , Neoplasias de las Glándulas Salivales/metabolismo , Glándulas Salivales/citología , Glándulas Salivales/embriología , Transducción de SeñalRESUMEN
For many years, our laboratory has been developing gene transfer approaches for salivary gland disorders that currently lack effective therapy. The purpose of this chapter is to describe key methods used in this developmental process. Specifically, we focus on one clinical condition, irradiation-induced salivary hypofunction, and address the choice of transgene and vector to be used, the construction of recombinant viral vectors, how vector delivery is accomplished, and methods for assessing vector function in vitro and in an appropriate animal model.
Asunto(s)
Terapia Genética/métodos , Enfermedades de las Glándulas Salivales/terapia , Adenoviridae/genética , Animales , Modelos Animales de Enfermedad , Vectores Genéticos/genética , Humanos , Ratones , Enfermedades de las Glándulas Salivales/genéticaRESUMEN
BACKGROUND: The purpose of this study was to examine the association between 4 Fas single nucleotide polymorphisms (SNPs) and risk of differentiated thyroid carcinoma (DTC) and salivary gland carcinoma (SGC). METHODS: We conducted a case-control study including 279 DTC cases, 165 benign thyroid disease (BTD) cases, 154 SGC cases, 61 benign salivary gland disease (BSGD) cases, and 510 controls. RESULTS: The A744G SNP genotype distribution was significantly different between subjects with SGC or BSGD and controls, while that of the A18272G SNP was significantly different between subjects with DTC or SGC and controls. Risk of SGC was significantly elevated for the 22628 heterozygous CT genotype (odds ratio [OR] = 1.5, p = .050), and risk of BSGD was elevated for the 22628 homozygous TT genotype (OR = 2.9, p = .023). CONCLUSION: Fas C22628T SNP may be associated with risk of SGC and BSGD, but none of the investigated Fas SNPs was associated with risk of DTC.
Asunto(s)
Carcinoma/genética , Polimorfismo de Nucleótido Simple , Neoplasias de las Glándulas Salivales/genética , Neoplasias de la Tiroides/genética , Receptor fas/genética , Estudios de Casos y Controles , Exones , Femenino , Predisposición Genética a la Enfermedad , Genotipo , Haplotipos , Humanos , Masculino , Persona de Mediana Edad , Regiones Promotoras Genéticas , Enfermedades de las Glándulas Salivales/genética , Enfermedades de la Tiroides/genéticaRESUMEN
DNA ploidy and S-Phase fraction (SPF) of 279 salivary gland tumours were analysed using high-resolution DNA flow cytometry. All 229 benign neoplasms were diploid while 12 of 50 malignant tumours showed cell populations with aneuploid DNA content. The SPF values of diploid malignancies were significantly higher if compared with pleomorphic adenomas but did not differ from that of the zystadenolymphoma (Warthin tumour) group. While aneuploidy represents a distinct indicator of malignancy SPF values are of minor relevance for dignity assessment in salivary gland tumours.
Asunto(s)
División Celular/genética , ADN de Neoplasias/genética , Ploidias , Neoplasias de las Glándulas Salivales/genética , Adenolinfoma/genética , Adenolinfoma/patología , Adenoma Pleomórfico/genética , Adenoma Pleomórfico/patología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Diagnóstico Diferencial , Diploidia , Femenino , Citometría de Flujo , Humanos , Masculino , Persona de Mediana Edad , Enfermedades de las Glándulas Salivales/genética , Enfermedades de las Glándulas Salivales/patología , Neoplasias de las Glándulas Salivales/patología , Glándulas Salivales/patología , Sensibilidad y EspecificidadRESUMEN
Recently, we reported development of the C57BL/6.NOD-Aec1Aec2 mouse carrying two genetic intervals derived from the NOD mouse. These two genetic regions confer full Sjögren's syndrome (SjS)-like disease in SjS-non-susceptible C57BL/6 mice. The current study was undertaken to apply microarray technology to define the molecular basis underlying onset of SjS-disease in C57BL/6.NOD-Aec1Aec2 mice. Using oligonucleotide microarrays, gene expression profiles of submandibular glands derived from 8- to 12-week-old C57BL/6.NOD-Aec1Aec2 mice and 8-week-old C57BL/6 mice were performed for comparison. Significant differential expressions were determined using the Mann-Whitney U test. Hybridizations using submandibular cDNA probes revealed 75 differentially expressed genes at 8 weeks and 105 differentially expressed genes at 12 weeks of age in C57BL/6.NOD-Aec1Aec2 mice compared to 8-week-old C57BL/6 mice. These genes were related generally to basic cellular activities such as transcription, translation, DNA replication, and protein folding. During the predisease phase, genes upregulated encode proteins associated with the IFN-gamma signal-transduction-pathway (Jak/Stat1), TLR-3 (Irf3 and Traf6) and apoptosis (casp11 and casp3), indicative of chronic proinflammatory stimuli, especially IL-1. Between 8 and 12 weeks of age, sets of clustered genes were upregulated that are associated with adaptive immune responses, especially B cell activation, proliferation and differentiation (Baffr, Taci, Bcma, Blys, April, CD70, CD40L, Traf1, Traf3, Pax5, c-Jun, Elk1 and Nf-kB), and neural receptors (Taj/Troy). Altered gene expressions of TLR3 and TNF-superfamily-receptors and ligands during this early phase of SjS suggest a possible viral etiology in the altered glandular homeostasis with an upregulated, possibly overstimulated, B-lymphocyte activation in the early autoimmune response present in the submandibular glands. The importance of NF-kappaB as a critical signal transduction pathway is also suggested but its link is not yet clear.
Asunto(s)
Enfermedades Autoinmunes/metabolismo , Enfermedades del Aparato Lagrimal/metabolismo , Enfermedades de las Glándulas Salivales/metabolismo , Síndrome de Sjögren/metabolismo , Glándula Submandibular/metabolismo , Factores de Edad , Animales , Enfermedades Autoinmunes/etiología , Enfermedades Autoinmunes/genética , Perfilación de la Expresión Génica , Enfermedades del Aparato Lagrimal/etiología , Enfermedades del Aparato Lagrimal/genética , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos NOD , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Enfermedades de las Glándulas Salivales/etiología , Enfermedades de las Glándulas Salivales/genética , Síndrome de Sjögren/etiología , Síndrome de Sjögren/genéticaRESUMEN
To date, the underlying genomic changes in benign and malignant tumors of salivary-gland and paranasal-sinus origin are poorly understood. This is due in part to the low incidence of these tumors and the enormous histological variety of tumors within this head and neck region. We examined 58 of these tumors (14 adenoid cystic carcinomas, 9 adenocarcinomas, 5 cylindrical carcinomas, 11 pleomorphic adenomas, and 19 inverted papillomas) by dual fluorescence in situ hybridization (FISH) with centromere-specific probes on six chromosomes (3, 7, 9, 11, 17, and 18) for numerical changes. In adenoid cystic carcinomas, monosomy of chromosome 17 and polysomy of chromosomes 3, 9 and 11 were most frequently encountered. In adenocarcinomas, monosomy of chromosome 17 and polysomy of chromosomes 7 and 11 were most frequent. In cylindrical cell carcinomas, polysomy of chromosomes 7, 9, 11 and 17 was present in the majority of tumors. Disomy is rare, even in benign tumors. Polysomy is more frequent in malignant tumors than in benign. Tetrasomy is found almost only in malignant tumors. In summary, the occurrence of polysomy might reflect a step towards malignancy in tumors of the salivary glands and paranasal mucosa. Polysomy of chromosome 11 could be defined as typical for all investigated histological types of malignant tumor in this region of the head and neck.
Asunto(s)
Adenocarcinoma/diagnóstico , Adenocarcinoma/genética , Adenoma/diagnóstico , Adenoma/genética , Carcinoma Adenoide Quístico/diagnóstico , Carcinoma Adenoide Quístico/genética , Aberraciones Cromosómicas , Papiloma/diagnóstico , Papiloma/genética , Neoplasias de las Glándulas Salivales/diagnóstico , Neoplasias de las Glándulas Salivales/genética , Diagnóstico Diferencial , Humanos , Hibridación Fluorescente in Situ , Enfermedades de las Glándulas Salivales/diagnóstico , Enfermedades de las Glándulas Salivales/genéticaRESUMEN
OBJECTIVE: To investigate the association between HLA antigens and temporomandibular joint (TMJ) erosion, salivary composition, and focal sialadenitis in patients with rheumatic diseases. METHODS: Eighty-four patients, 24 with rheumatoid arthritis (RA), 19 with mixed connective tissue disease (MCTD), 19 with ankylosing spondylitis (AS), and 22 with spondyloarthropathy (SPA) were studied. Each patient underwent clinical examination of the masticatory system, unstimulated and stimulated saliva collection, and minor salivary gland biopsy. Radiographs (OPTG) of the TMJ were obtained, and HLA allele (A, B, C and DRB1*) analysis was performed. Erosion in OPTG was scored from 0 (no erosion) to 4 (condyles totally eroded). In the analysis, scores 0-2 were grouped as normal or mild changes, and scores 3-4 as distinct erosions. One hundred healthy blood donors served as controls for HLA typing. RESULTS: Distinct erosion of the TMJ in OPTG was observed in 22 (27%) patients. It affected four (17%) of the 24 patients with RA, three (17%) of the 18 with MCTD, seven (37%) of the 19 patients with AS and eight (38%) of the 21 with SPA non-significant (NS). The mean erosion scores were 1.7 for RA, 1.3 for MCTD, 2.5 for SPA, and 1.6 for AS patients [probability (p) = 0.04]. The frequency of HLA-B27 antigen was higher in the AS and SPA patients, and that of HLA-DRB1*04 allele higher in RA patients than in control subjects. In the whole patient population, HLA-DRB1*01 allele was significantly associated with erosions 16/36 (44%) versus 6/46 (131%1) (p = 0.0014). In the SPA group, patients with HLA-DRBI*01 allele had a significantly higher occurrence of distinct erosions than patients without this allele [8/10 (80%) versus 0/11 (0%) (p = 0.0002)], whereas DRB1*06 was protective [0/8 (0%) versus 8/13 (62%) (p = 0.018)]. HLA-DRB1*04 was associated with increased salivary IgG in the RA patients. CONCLUSION: HLA antigens are significantly associated with the development of destructive lesions in the TMJ, as well as composition of saliva in patients with various rheumatic diseases.
Asunto(s)
Predisposición Genética a la Enfermedad , Antígenos HLA-DR/genética , Enfermedades Reumáticas/genética , Enfermedades de las Glándulas Salivales/genética , Trastornos de la Articulación Temporomandibular/genética , Alelos , Artritis Reumatoide/epidemiología , Artritis Reumatoide/genética , Estudios de Cohortes , Enfermedades del Tejido Conjuntivo/epidemiología , Enfermedades del Tejido Conjuntivo/genética , Femenino , Finlandia/epidemiología , Regulación de la Expresión Génica , Cadenas HLA-DRB1 , Humanos , Masculino , Reacción en Cadena de la Polimerasa , Prevalencia , Pronóstico , Estudios Prospectivos , Radiografía , Enfermedades Reumáticas/epidemiología , Medición de Riesgo , Enfermedades de las Glándulas Salivales/epidemiología , Sensibilidad y Especificidad , Índice de Severidad de la Enfermedad , Espondilitis Anquilosante/epidemiología , Espondilitis Anquilosante/genética , Trastornos de la Articulación Temporomandibular/diagnóstico por imagen , Trastornos de la Articulación Temporomandibular/epidemiologíaRESUMEN
BACKGROUND: Localized gene transfer to salivary glands has great potential for the treatment of salivary gland, systemic, and oral diseases. The minipig parotid gland, given its volume and morphological similarities to the human parotid gland, may be useful as a large animal model for pre-clinical gene transfer experiments. The purpose of this study was to perform an initial assessment of the efficacy and safety of adenoviral-vector-mediated gene transfer to parotid glands of miniature pigs. METHODS: AdCMVluc, a recombinant type 5 adenoviral (rAd5) vector containing a luciferase reporter gene, was administered to miniature pig parotid glands by intraductal cannulation. Five regions of gland tissue were obtained to measure the distribution of luciferase activity. The effects of time, viral dose, infusate buffer volume, and gland anatomical region on transgene expression were determined. Detailed serum chemistry and hematological analyses were performed. In addition, AdCMVlacZ, a similar rAd5 vector encoding beta-galactosidase, was also delivered to determine the parotid gland cell types transduced. RESULTS: Luciferase assays indicated that gene transfer to miniature pig salivary glands could be readily accomplished using rAd5 vectors. Highest transgene expression was found in the center of glands, which was > posterior > inferior > anterior > superior tissue regions. Expression was maximal on day 2 and declined to background by day 14, and observed in both acinar and ductal cells. Several serum chemistry and hematology parameters were transiently changed following rAd5 administration. CONCLUSIONS: Transgene expression by, and inflammatory response to, rAd5 vectors in minipig parotid glands are similar to results seen earlier in rodent studies. This suggests that results of salivary gland gene transfer from rodent studies can be extended to a larger animal model, and supports the value of using minipigs for pre-clinical applications of gene transfer to these tissues. Published in 2004 by John Wiley & Sons, Ltd.
Asunto(s)
Adenoviridae/genética , Perfilación de la Expresión Génica , Técnicas de Transferencia de Gen , Luciferasas/genética , Glándula Parótida , Porcinos Enanos/genética , Animales , Genes Reporteros , Vectores Genéticos , Inflamación , Luciferasas/biosíntesis , Masculino , Modelos Animales , Enfermedades de las Glándulas Salivales/genética , Enfermedades de las Glándulas Salivales/terapia , PorcinosRESUMEN
The roles of apoptosis and mitosis of acinar and duct cells in the atrophy of the sublingual gland of rat induced by double duct ligation was investigated using immunohistochemistry for proliferating cell nuclear antigen (PCNA), terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-digoxigenin nick end labeling (TUNEL), and transmission electron microscopy (TEM). Many PCNA-positive duct cells were observed 3 days after duct ligation, and the numbers decreased thereafter. At 3 and 5 days, several TUNEL-positive acinar cells were observed and typical apoptotic acinar cells were identified by TEM. Necrotic acinar cells were also observed ultrastructurally. After 7 days, there were few acini but many ducts, as well as many structures representing transition from acinus to duct. These observations demonstrate that acinar cell loss by apoptosis and duct cell proliferation by mitosis occur in atrophic sublingual glands as well as in other atrophic salivary glands. In addition, it appears that the transition from acinar to duct cell and the necrosis of acinar cells play important roles in the atrophy of the sublingual gland.
Asunto(s)
Apoptosis/fisiología , Atrofia/metabolismo , Mitosis/fisiología , Enfermedades de las Glándulas Salivales/metabolismo , Glándula Sublingual/metabolismo , Animales , Atrofia/genética , Atrofia/patología , Núcleo Celular/patología , Núcleo Celular/ultraestructura , Citoplasma/patología , Citoplasma/ultraestructura , Células Epiteliales/metabolismo , Células Epiteliales/patología , Células Epiteliales/ultraestructura , Etiquetado Corte-Fin in Situ , Masculino , Microscopía Electrónica , Necrosis , Orgánulos/patología , Orgánulos/ultraestructura , Antígeno Nuclear de Célula en Proliferación/metabolismo , Ratas , Ratas Wistar , Enfermedades de las Glándulas Salivales/genética , Enfermedades de las Glándulas Salivales/patología , Glándula Sublingual/patología , Glándula Sublingual/fisiopatologíaRESUMEN
BACKGROUND: Gene therapy is an emerging field of biomedicine that has commanded considerable scientific and popular attention. The procedure involves the transfer of genes to patients for clinical benefit. Transferred genes can b e used for either reparative or pharmacological purposes. OVERVIEW: In 1995, the first author and a colleague described the potential impact of gene therapy on dentistry, on the basis of initial studies of gene transfer applications to salivary glands, keratinocytes and cancer cells. Their conclusion was that gene therapy would have a significant impact on the nature of dental practice within 20 years. In this article, the authors consider research progress since 1995 and reexamine the earlier conclusion. PRACTICE IMPLICATIONS: In the past six years, remarkable progress has been made in the field of gene therapy, including seven areas relevant to dental practice: bone repair, salivary glands, autoimmune disease, pain, DNA vaccinations, keratinocytes and cancer. While considerable problems remain, thus impeding the routine clinical use of gene transfer, gene therapy will have a pervasive and significant impact on areas of dental practice that are based in biological science. By 2015, this will translate into practitioners' having a wide range of novel biological treatment options for their patients.
Asunto(s)
Terapia Genética , Enfermedades Autoinmunes/terapia , Proteínas Morfogenéticas Óseas/genética , Dolor Facial/terapia , Técnicas de Transferencia de Gen , Neoplasias de Cabeza y Cuello/terapia , Humanos , Queratinocitos/metabolismo , Enfermedades de la Boca/terapia , Enfermedades de las Glándulas Salivales/genética , Síndrome de Sjögren/terapia , Enfermedades Dentales/terapia , Vacunación , Vacunas de ADNRESUMEN
Considerable progress has been made in the transfer of foreign genes into salivary glands in vivo using adenovirus vectors in rats. In an attempt to avoid the transient expression inherent, when using these vectors, retroviral vectors and human cell lines where used here in attempt to develop an in vitro model of HIV-associated salivary gland disease. The HIV-1-tat protein is increasingly implicated in the pathogenesis of the AIDS through altering the expression of strategic cellular genes. The purpose of this study was to transfect human salivary gland (HSG) cell lines in vitro, with the pHIV-1/LTR-tat plasmid, and examine the effect of tat on expression of matrix and basement membrane genes known to be important in the pathogenesis of salivary gland disease. HSG cells were transfected with HIV-1-tat plasmid by the lipofection method. Transfection was confirmed by polymerase chain reaction (PCR) and Southern blot, which verified that tat-specific DNA was present. Tat-mRNA was analysed by Northern blotting and quantified by reverse transcriptase polymerase chain reaction (RT-PCR) to demonstrate its expression. Numerous clones were found to contain integrated tat DNA sequences and analysis of mRNA showed stable expression of tat-specific RNA. Further analysis of mRNA expression for various marker proteins important in HIV pathogenesis showed that the HSG cell line transfected with HIV-1-tat, was associated with significant induction of mRNA expression for extracellular matrix protein. Tat-amplified transcription of the major basement membrane protein laminin, as well as of fibronectin, collagen I and III, and c-myc oncogene was demonstrated. Conversely, expression of p53 suppressor gene mRNA was reduced. Post-transfection expression of collagen IV was erratic and inconclusive. It was concluded that the presence of HIV-tat in this in vitro model of salivary ductal epithelial cell model alters the mRNA expression of several matrix, basement membrane and oncoproteins known to be involved in HIV pathogenesis. These cell lines provide a useful system for studying the role of tat in the immunopathogenesis of HIV-associated salivary gland disease.
Asunto(s)
Colágeno/genética , Proteínas de la Matriz Extracelular/genética , Fibronectinas/genética , Amplificación de Genes , Genes myc/genética , Genes p53/genética , Genes tat/genética , VIH/genética , Laminina/genética , Oncogenes/genética , Glándulas Salivales/metabolismo , Transfección , Membrana Basal/metabolismo , Southern Blotting , Línea Celular , ADN Viral/análisis , Células Epiteliales/metabolismo , Matriz Extracelular/metabolismo , Regulación Viral de la Expresión Génica , Infecciones por VIH/genética , Humanos , Plásmidos , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Conductos Salivales/metabolismo , Enfermedades de las Glándulas Salivales/genética , Enfermedades de las Glándulas Salivales/virología , Transcripción GenéticaRESUMEN
MRL/MpJ-lpr/lpr (MRL/lpr) mice develop collagen disease involving vasculitis, glomerulonephritis, arthritis and sialoadenitis, each of which has been studied as a model for polyarteritis, lupus nephritis, rheumatoid arthritis and Sjögren's syndrome, respectively. In the previous studies, we observed genetic segregation of these complex pathological manifestations throughout the genome recombination with a C57Bl/6-lpr/lpr or a C3H/HeJ-lpr/lpr (C3H/lpr) strain of mice which rarely develops such lesions, indicating that development of collagen disease is dependent on an MRL host genetic background. To clarify the mode of inheritance and the gene loci affecting four types of the lesions in MRL/lpr mice; vasculitis, glomerulonephritis, arthritis and sialoadenitis, a genetic dissection of the lesions was carried out by using MRL/lpr, C3H/lpr, (MRL/lprxC3H/lpr) F1 intercross, and MRL/lprx(MRL/lprxC3H/lpr) F1 backcross mice. Definition of each lesion was performed by histopathology under light microscopy, and genomic DNA of the backcross mice were subjected to association studies by chi-square analysis for determining which polymorphic microsatellite locus occurs at higher frequency among affected compared to unaffected individuals for each lesion. We observed that gene loci recessively associated with each lesion were mapped on different chromosomal positions. We conclude that each of four types of the lesions in MRL/lpr mice is under the control of different set of genes, suggesting the complex pathological manifestations of collagen disease result from polygenic inheritance.
Asunto(s)
Artritis/genética , Enfermedades del Colágeno/genética , Glomerulonefritis/genética , Enfermedades de las Glándulas Salivales/genética , Vasculitis/genética , Animales , Artritis/patología , Segregación Cromosómica , Enfermedades del Colágeno/patología , Ligamiento Genético , Glomerulonefritis/patología , Ratones , Ratones Endogámicos MRL lpr , Enfermedades de las Glándulas Salivales/patología , Vasculitis/patologíaRESUMEN
Recombinant DNA technology is finding its way into many aspects of clinical medicine. No application currently is more dramatic than gene therapy. Proofs of principle have already been established for gene therapy targeted to oral tissues, and more are likely to be demonstrated in the near future. The dental curriculum must begin to include the biological basis of DNA-based therapies and other related biomedical science progress.
Asunto(s)
Educación en Odontología , Técnicas de Transferencia de Gen , Genética Médica/educación , Enfermedades de las Glándulas Salivales/terapia , Curriculum , Humanos , Enfermedades de las Glándulas Salivales/genéticaRESUMEN
Microdissection of biopsy material from labial minor salivary glands followed by RT-PCR demonstrates extensive production of cytokines IL-2, 1L-6, IL-10, TNF-alpha, TGF-beta, and IFN-gamma in both normal glands and in glands affected by Sjögren's syndrome. Continuation of these studies should expand our knowledge of normal salivary gland function and the pathogenesis of Sjögren's syndrome.