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Eduardo M Rocha, Changyu Zheng, Lara Dias, Luis Fernando Nominato, Ana Carolina Dias; Adenoviral vector-mediated transfer of erythropoietin in the lacrimal gland. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):3477.
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© ARVO (1962-2015); The Authors (2016-present)
The aims of this work were to analyze the alterations in the lacrimal gland (LG) and ocular surface (OS) of rats transfected in the LG with an adenovirus (Ad) encoding the human erythropoietin (Epo) gene (AdLTR2EF1a-hEPO).
Male Wistar rats 6 weeks-old were divided into 2 groups (n = 5/group), where the control group received an injection of saline and AdEpo received an injection of AdLTR2EF1a-hEPO (25ul, 1010 particles/ml), in the LG. Data was collected 6 days after injection. It was evaluated tears secretion (phenol red thread test), OS impression cytology (IC), LG and cornea histology, quantification of hEPO mRNA by qRT-PCR and protein by western blotting (WB), Elisa and location by immunohistochemistry.
At the 6 th day after injection, tear secretion was not affected by viral vector gene transfer (control 7.4 ± 1.14mm, AdEpo 7.0± 1.6 mm, p=0.66). There was no change in LG histology compared to controls. The cornea epithelial thickness was 34.3± 3.7µm in the control, 31.8±3.8µm in the AdEPO group (p=0.099). hEPO mRNA was positive only in the AdEPO group. WB hEpo quantification was significantly higher in transfected LG than controls (p<0.0001). hEPO location was positive in LG ductal cells of AdEPO group.
AdEpo was safe for LG structure and function. It increased the local expression of Epo. The present work offers additional information on the feasibility of gene therapy in the LG.
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