Establishment of a human corneal epithelial cell line lacking the functional TACSTD2 gene as an in vitro model for gelatinous drop-like dystrophy.

Abstract

PURPOSE:

Gelatinous drop-like corneal dystrophy (GDLD) is characterized by subepithelial amyloid deposition that engenders severe vision loss. The exact mechanism of this disease has yet to be elucidated. No fundamental treatment exists. This study was conducted to establish an immortalized corneal epithelial cell line to be used as a GDLD disease model.

METHODS:

A corneal tissue specimen was obtained from a GDLD patient during surgery. Corneal epithelial cells were enzymatically separated from the cornea and were dissociated further into single cells. The epithelial cells were immortalized by the lentiviral transduction of the simian virus 40 (SV40) large T antigen and human telomerase reverse transcriptase (hTERT) genes. For the immortalized cells, proliferative kinetics, gene expressions, and functional analyses were performed.

RESULTS:

The immortalized corneal epithelial cells continued to proliferate despite cumulative population doubling that exceeded 100. The cells showed almost no sign of senescence and displayed strong colony-forming activity. The cells exhibited a low epithelial barrier function as well as decreased expression of tight-junction-related proteins claudin 1 and 7. Using the immortalized corneal epithelial cells derived from a GDLD patient, we tested the possibility of gene therapy.

CONCLUSIONS:

We established an immortalized corneal epithelial cell line from a GDLD patient. The immortalized cells exhibited cellular phenotypes similar to those of in vivo GDLD. The immortalized cells are thought to be useful for the development of new therapies for treating GDLD corneas and for elucidation of the pathophysiology of GDLD.

• Authors: Koji Kitazawa, Satoshi Kawasaki, Katsuhiko Shinomiya, Keita Aoi, Akira Matsuda, Toshinari Funaki, Kenta Yamasaki, Mina Nakatsukasa, Nobuyuki Ebihara, Akira Murakami, Junji Hamuro, Shigeru Kinoshita
• Journal: Investigative ophthalmology & visual science (Invest Ophthalmol Vis Sci) Vol. 54 Issue 8 Pg. 5701-11 (Aug 23 2013) ISSN: 1552-5783 [Electronic] United States
• PMID: 23868985 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, Neoplasm
• Cell Adhesion Molecules
• TACSTD2 protein, human
• RNA

Topics

• Amyloidosis, Familial (genetics, metabolism, pathology)
• Antigens, Neoplasm (genetics, metabolism)
• Blotting, Western
• Cell Adhesion Molecules (genetics, metabolism)
• Cell Line
• Cell Proliferation
• Corneal Dystrophies, Hereditary (genetics, metabolism, pathology)
• Epithelium, Corneal (metabolism, pathology)
• Female
• Gene Expression Regulation
• Humans
• Middle Aged
• RNA (genetics)
• Reverse Transcriptase Polymerase Chain Reaction