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Koji Kitazawa, Satoshi Kawasaki, Keita Aoi, Katsuhiko Shinomiya, Akira Matsuda, Toshinari Funaki, Mina Nakatsukasa, Junji Hamuro, Akira Murakami, Shigeru Kinoshita; Investigation of Gene Therapy Using Immortalized Cells Derived from a Gelatinous Drop-Like Corneal Dystrophy Patient. Invest. Ophthalmol. Vis. Sci. 2013;54(15):4559. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Gelatinous drop-like corneal dystrophy (GDLD) is an autosomal recessive inheritance disease caused by biallelic loss-of-function mutations within the tumor-associated calcium signal transducer 2 (TACSTD2) gene. We previously established two immortalized cell lines via the lentiviral introduction of the SV40 large T antigen and hTERT genes into the corneal and conjunctival epithelial cells of GDLD patients. The purpose of this present study was to assess whether or not a gene therapy using these two cells lines is effective for the treatment of GDLD patients.
The lentivirus vector harboring the wild-type TACSTD2 gene was produced and transduced to the two immortalized cell lines described above. Epithelial barrier function of the two cell lines was investigated by means of trans-epithelial resistance (TER) analysis and dye permeabilization testing using fluorescein in order to assess whether or not the transduction of the wild-type TACSTD2 gene actually normalizes the disease situation of GDLD cornea.
The transduction efficiency of the wild-type TACSTD2 gene to the immortalized cell lines was approximately 80%. TER of the immortalized cell lines was increased after the transduction of the wild-type TACSTD2 gene. The permeability of fluorescein in the immortalized cell lines was decreased after the transduction of the wild-type TACSTD2 gene.
The findings of this study show that transduction of the wild-type TACSTD2 gene normalizes the disease situation of GDLD corneas to some extent, thus indicating that gene therapy may prove to be a promising treatment for GDLD.
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