Transient adenoviral gene transfer of Smad7 prevents injury-induced epithelial–mesenchymal transition of lens epithelium in mice

We examined the effect of adenovirus-mediated transient expression of Smad7, an inhibitory Smad in TGFβ/activin signaling, on injury-induced epithelial–mesenchymal transition (EMT) of lens epithelium in mice. A volume of 3 μl of adenoviral solution was injected into the right lens of adult male C57B...

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Published in:Laboratory investigation 2004-10, Vol.84 (10), p.1259-1270
Main Authors: Saika, Shizuya, Ikeda, Kazuo, Yamanaka, Osamu, Sato, Misako, Muragaki, Yasuteru, Ohnishi, Yoshitaka, Ooshima, Akira, Nakajima, Yuji, Namikawa, Kazuhiko, Kiyama, Hiroshi, Flanders, Kathleen C, Roberts, Anita B
Format: Article
Language:English
Subjects:
Actins - metabolism
Adenoviridae - genetics
Animals
Biological and medical sciences
Biotechnology
Cell Division
Chondroitin Sulfate Proteoglycans - metabolism
Collagen Type VI - metabolism
Disease Models, Animal
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Epithelial Cells - metabolism
Epithelial Cells - pathology
epithelial–mesenchymal transition
Eye Injuries - prevention & control
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation
Genetic Therapy
Genetic Vectors - genetics
Investigative techniques, diagnostic techniques (general aspects)
Keratan Sulfate - metabolism
Lens Capsule, Crystalline - injuries
Lens Capsule, Crystalline - metabolism
Lens Capsule, Crystalline - pathology
lens epithelial cell
Lens, Crystalline - metabolism
Lens, Crystalline - pathology
Lumican
Male
Medical sciences
Mesoderm - pathology
Mesoderm - physiology
Mice
Mice, Inbred C57BL
mouse
RNA, Messenger - metabolism
Smad
Smad7 Protein
Trans-Activators - genetics
Trans-Activators - metabolism
Transduction, Genetic
Transforming Growth Factor beta - genetics
Transforming Growth Factor beta - metabolism
transforming growth factor β
Wound Healing
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Summary:We examined the effect of adenovirus-mediated transient expression of Smad7, an inhibitory Smad in TGFβ/activin signaling, on injury-induced epithelial–mesenchymal transition (EMT) of lens epithelium in mice. A volume of 3 μl of adenoviral solution was injected into the right lens of adult male C57BL/6 mice (n=56) at the time of capsular injury made using a hypodermic needle under general anesthesia. A mixture of recombinant adenovirus carrying CAG promoter-driven Cre (Cre adv) and mouse Smad7 complementary DNA (Smad7 adv) was administered to induce Smad7 expression, while control lenses were treated with Cre adv alone. After healing intervals of 2, 3, 5, and 10 days, animals were killed 2 h after labeling with bromodeoxyuridine (BrdU) and eyes were processed for histology. During healing, marked expression of Smad7 was observed in lens epithelial cells in the Smad7 adv group with loss of nuclear translocation of Smads2/3, while little Smad7 and abundant nuclear Smads2/3 were seen in cells in the Cre adv group. Lens epithelial cells in the Cre adv control group exhibited a fibroblastic appearance at days 5 and 10 and the capsular break was sealed with fibrous tissue, while Smad7 adv-treated cells around the capsular break retained their epithelial morphology and the break was not sealed. Expression of snail mRNA, and α-smooth muscle actin, lumican, and collagen VI proteins, markers of EMT, was observed in control-treated eyes, but not in cells of the Smad7 adv group at day 5 with minimal expression at day 10. Additionally, cell proliferation increased in epithelium infected with Smad7 adv consistent with suppression of injury-induced upregulation of TGFβ1 in epithelium. We conclude that gene transfer of Smad7 in mice prevents injury-induced EMT of lens epithelial cells and sealing of the capsular break with fibrous tissue.
ISSN:0023-6837
1530-0307
DOI:10.1038/labinvest.3700151