June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Adenoviral vector-mediated transfer of erythropoietin in the lacrimal gland
Author Affiliations & Notes
  • Eduardo M Rocha
    Ophthalmology, FMRP-USP, Ribeirao Preto, Brazil
  • Changyu Zheng
    NIDCR/NIH, Bethesda, MD
  • Lara Dias
    Ophthalmology, FMRP-USP, Ribeirao Preto, Brazil
  • Luis Fernando Nominato
    Ophthalmology, FMRP-USP, Ribeirao Preto, Brazil
  • Ana Carolina Dias
    Ophthalmology, FMRP-USP, Ribeirao Preto, Brazil
  • Footnotes
    Commercial Relationships Eduardo Rocha, None; Changyu Zheng, None; Lara Dias, None; Luis Fernando Nominato, None; Ana Dias, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 3477. doi:
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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)

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Abstract

Purpose: 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).

Methods: 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.

Results: 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.

Conclusions: 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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