Investigation of Gene Therapy Using Immortalized Cells Derived from a Gelatinous Drop-Like Corneal Dystrophy Patient

Abstract

Purpose: 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.

Methods: 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.

Results: 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.

Conclusions: 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.

Keywords: 482 cornea: epithelium • 494 degenerations/dystrophies • 538 gene transfer/gene therapy