PubMed Health⌕ Search
Home/Archive/PubMed

PubMed · 16604526

Transgenic mouse for conditional, tissue-specific Cox-2 overexpression.

Ken-ichiro Kamei·Tomo-o Ishikawa·Harvey R Herschman
View original source ↗Export metadata ↓

Abstract

We constructed a cyclooxygenase-2 (Cox-2) conditional overexpression transgenic mouse (Cox-2 COE). The transgene contains a CAG promoter driving the Cox-2 and humanized Renilla luciferase (hRL) coding regions, linked by an internal ribosomal entry site. The promoter is followed by a loxP-flanked sequence containing enhanced green fluorescent protein (EGFP), a neomycin selection cassette, and a transcriptional/translational STOP sequence. In the presence of Cre recombinase the loxP-flanked sequence is excised. Cox-2/hRL expression can be monitored repeatedly and noninvasively in vivo by imaging hRL activity. To demonstrate conditional Cox-2 and hRL expression, a nonreplicating adenovirus carrying Cre recombinase (Ad.CMV.Cre) was injected intravenously; hepatic Cox-2 expression and hRL signal were elevated. Cox-2 COE embryonic fibroblasts express both Cox-2 and hRL following Ad.CMV.Cre infection. PGE(2) production is also increased following Ad.CMV.Cre infection of Cox-2 COE embryo fibroblasts. Cox-2 COE mice should be valuable for the study of Cox-2 overexpression in cardiovascular disease, acute and chronic inflammatory responses, neurodegenerative diseases, and cancer.

Explore related subjects

AdenoviridaeAnimalsCyclooxygenase 2FibroblastsGene DeletionGene Expression RegulationGenetic TechniquesGenetic VectorsGreen Fluorescent ProteinsInjections, IntravenousIntegrasesLiverLuciferases, RenillaMiceMice, TransgenicModels, GeneticPromoter Regions, GeneticTransgenesViral ProteinsJournal ArticleResearch Support, N.I.H., Extramural

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Ken-ichiro Kamei, Tomo-o Ishikawa, Harvey R Herschman. 2006. Transgenic mouse for conditional, tissue-specific Cox-2 overexpression.. https://doi.org/10.1002/dvg.20199

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed2007

Periostin regulates collagen fibrillogenesis and the biomechanical properties of connective tissues.

Periostin is predominantly expressed in collagen-rich fibrous connective tissues that are subjected to constant mechanical stresses including: heart valves, tendons, perichondrium, cornea, and the periodontal ligament (PDL). Based on these data we hypothesize that periostin can regulate collagen I fibrillogenesis and thereby affect the biomechanical properties of connective tissues. Immunoprecipitation and immunogold transmission electron microscopy experiments demonstrate that periostin is capable of directly interacting with collagen I. To analyze the potential role of periostin in collagen I fibrillogenesis, gene targeted mice were generated. Transmission electron microscopy and morphometric analyses demonstrated reduced collagen fibril diameters in skin dermis of periostin knockout mice, an indication of aberrant collagen I fibrillogenesis. In addition, differential scanning calorimetry (DSC) demonstrated a lower collagen denaturing temperature in periostin knockout mice, reflecting a reduced level of collagen cross-linking. Functional biomechanical properties of periostin null skin specimens and atrioventricular (AV) valve explant experiments provided direct evidence of the role that periostin plays in regulating the viscoelastic properties of connective tissues. Collectively, these data demonstrate for the first time that periostin can regulate collagen I fibrillogenesis and thereby serves as an important mediator of the biomechanical properties of fibrous connective tissues.

Adenoviridae↗
PubMed2007

SUMF1 enhances sulfatase activities in vivo in five sulfatase deficiencies.

Sulfatases are enzymes that hydrolyse a diverse range of sulfate esters. Deficiency of lysosomal sulfatases leads to human diseases characterized by the accumulation of either GAGs (glycosaminoglycans) or sulfolipids. The catalytic activity of sulfatases resides in a unique formylglycine residue in their active site generated by the post-translational modification of a highly conserved cysteine residue. This modification is performed by SUMF1 (sulfatase-modifying factor 1), which is an essential factor for sulfatase activities. Mutations in the SUMF1 gene cause MSD (multiple sulfatase deficiency), an autosomal recessive disease in which the activities of all sulfatases are profoundly reduced. In previous studies, we have shown that SUMF1 has an enhancing effect on sulfatase activity when co-expressed with sulfatase genes in COS-7 cells. In the present study, we demonstrate that SUMF1 displays an enhancing effect on sulfatases activity when co-delivered with a sulfatase cDNA via AAV (adeno-associated virus) and LV (lentivirus) vectors in cells from individuals affected by five different diseases owing to sulfatase deficiencies or from murine models of the same diseases [i.e. MLD (metachromatic leukodystrophy), CDPX (X-linked dominant chondrodysplasia punctata) and MPS (mucopolysaccharidosis) II, IIIA and VI]. The SUMF1-enhancing effect on sulfatase activity resulted in an improved clearance of the intracellular GAG or sulfolipid accumulation. Moreover, we demonstrate that the SUMF1-enhancing effect is also present in vivo after AAV-mediated delivery of the sulfamidase gene to the muscle of MPSIIIA mice, resulting in a more efficient rescue of the phenotype. These results indicate that co-delivery of SUMF1 may enhance the efficacy of gene therapy in several sulfatase deficiencies.

Adenoviridae↗
PubMed2007

Gene transfer of the CD40-ligand to human dendritic cells induces NK-mediated antitumor effects against human carcinoma cells.

The CD40-ligand (CD40L) is a key molecule for the activation of dendritic cells (DCs), followed by the induction of DC maturation and cytokine production. Here we found that DC infected with adenovirus vector encoding human CD40L (CD40L-DC) displayed significantly higher levels of immune accessory molecules and IL-12 production than did uninfected cells, and that CD40L-DC produced much higher levels of IFN-gamma. To investigate whether CD40L-DC-derived these soluble factors could stimulate NK cells without physical cell-to-cell contact, we cocultured NK cells with CD40L-DC in transwell culture plates. NK cells showed up-regulated cytotoxic activity toward various squamous oral cell carcinoma (OSC-70, HSC-2, HSC-3), and we determined that both IL-12 and IFN-gamma contributed to the CD40L-DC-mediated NK cell activation. NK cells stimulated with CD40L-DC resulted in the induction of the cell surface expression of TRAIL, the production of IFN-gamma and intracellular accumulation of granzyme B. The cytotoxic activity of NK cells stimulated with CD40L-DC could be mostly inhibited by neutralizing antibody for TRAIL and completely abrogated by the combination of antibody and exocytosis inhibitor, indicating that this was mainly mediated by a TRAIL-TRAIL-receptor interaction and granule exocytosis. Moreover, CD40L-DC-activated NK cells could induce up-regulation of a death-receptor TRAIL-R2 (DR5) and down-regulation of a decoy receptor TRAIL-R3 (DcR1) on carcinoma cells. Overall, these results have revealed that CD40L-DC could activate an innate immune reaction by stimulating NK cells followed by carcinoma cells, supporting that administration of CD40L-DC may have potential as an anticancer therapy.

Adenoviridae↗