Adenoviral gene transfer of BMP-7, Id2, or Id3 suppresses injury-induced epithelial-to-mesenchymal transition of lens epithelium in mice.
We have examined the effect of adenovirus-mediated expression of bone morphogenic protein-7 (BMP-7) and inhibitors of differentiation 2 and 3 (Id2 and Id3) on injury-induced epithelial-to-mesenchymal transition (EMT) of lens epithelium in mice. Id2 and Id3 are known to be upregulated by BMP-7 and to antagonize Smad2/3 signaling. The Cre-LoxP system adenoviral gene transfer was used. Three microliters of adenoviral solution (2 x 10(7) PFU/mul) were injected into the right lens of adult male C57BL/6 mice (n = 144) at the time of capsular injury induced using a hypodermic needle under both general and topical anesthesia. A mixture of Cre-adenovirus (Cre-Ad) and vector encoding mBMP-7, mId2, or mId3 was administered in a test group. Control lenses were treated with Cre-Ad alone. After healing intervals of 5 or 10 days, the animals were killed and then we performed histological processes or RNA extraction from the lens. RT-PCR, real-time RT-PCR, and immunohistochemistry showed expression of each introduced gene in the lens. Exogenous BMP-7 upregulated expression of Id2 and Id3 in injured lenses, and gene introduction of Id2 or Id3 also upregulated BMP-7 expression. Gene transfer of BMP-7, Id2, or Id3 delayed injury-induced EMT of the lens epithelial cells as evaluated by histology and expression patterns of alpha-smooth muscle actin and collagens in association with reduction of Smad2 COOH-terminal phosphorylation. Gene transfer of BMP-7, Id2, or Id3 delayed injury-induced EMT of lens epithelial cells and subsequent sealing of the capsular break with fibrous tissue in mice.
Saika
Shizuya
S
Dept. of Ophthalmology, Wakayama Medical Univ., 811-1 Kimiidera, Wakayama 641-0012, Japan. shizuya@wakayama-med.ac.jp
Ikeda
Kazuo
K
Yamanaka
Osamu
O
Flanders
Kathleen C
KC
Ohnishi
Yoshitaka
Y
Nakajima
Yuji
Y
Muragaki
Yasuteru
Y
Ooshima
Akira
A
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015415:Biomarkers; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins / Q000235:genetics* / Q000378:metabolism; D002386:Cataract / Q000473:pathology / Q000503:physiopathology / Q000628:therapy; D004847:Epithelial Cells / Q000166:cytology; D005355:Fibrosis; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D051797:Inhibitor of Differentiation Protein 2 / Q000235:genetics* / Q000378:metabolism; D051796:Inhibitor of Differentiation Proteins / Q000235:genetics* / Q000378:metabolism; D007908:Lens, Crystalline / Q000293:injuries* / Q000473:pathology* / Q000502:physiology; D008648:Mesoderm / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D010766:Phosphorylation; D051785:Smad Proteins / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000235:genetics* / Q000378:metabolism
D015415:Biomarkers; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins; C497452:Idb2 protein, mouse; D051797:Inhibitor of Differentiation Protein 2; D051796:Inhibitor of Differentiation Proteins; D051785:Smad Proteins; D016212:Transforming Growth Factor beta; C067679:Idb3 protein, mouse