Adenoviral gene transfer of bioactive TGFβ1 to the rodenteye as a novel model for anterior subcapsular cataract

[摘要] Purpose: To produce a gene-transfer model of rodent anteriorsubcapsular cataracts (ASC) using a replication-deficient, adenoviralvector containing active TGFβ1. Establishment of this model will beimportant for further investigations of TGFβ-induced signalingcascades in ASC.Methods: Adenovirus containing the transgene for active TGFβ1(AdTGFβ1), β-galactosidase (AdLacZ), green fluorescent protein(AdGFP) or no transgene (AdDL) was injected into the anterior chamber ofC57Bl/6, Smad3 WT and Smad3 KO mice. Four and 21 days after injection,animals were enucleated and eyes were processed and examined by routinehistology. Immunolocalization of markers indicative of epithelial tomesenchymal transition (EMT), fibrosis, proliferation and apoptosis wasalso carried out.Results: By day 4, treatment with AdLacZ demonstrated transgeneexpression in multiple structures of the anterior chamber including thelens epithelium. In contrast to AdDL, treatment with AdTGFβ1produced αSMA-positive subcapsular plaques in all three groups ofmice, which shared features reminiscent of human ASC. At day 21, plaquesremained αSMA-positive and extensive extracellular matrixdeposition was observed. The AdTGFβ1 model was further employed inSmad3 deficient mice and this resulted in the development of small ASC.Conclusions: Gene transfer of active TGFβ1 using an adenoviralvector produced cataractous plaques four days postinjection, which werefound to develop independent of functional Smad3.

[学科分类] 生物化学/生物物理