TY - JOUR
T1 - Effect of trinucleotide repeat expansion on the expression of TCF4 mRNA in fuchs’ endothelial corneal dystrophy
AU - Okumura, Naoki
AU - Hayashi, Ryosuke
AU - Nakano, Masakazu
AU - Yoshii, Kengo
AU - Tashiro, Kei
AU - Sato, Takahiko
AU - Blake, Derek J.
AU - Aleff, Ross
AU - Butz, Malinda
AU - Highsmith, Edward W.
AU - Wieben, Eric D.
AU - Fautsch, Michael P.
AU - Baratz, Keith H.
AU - Komori, Yuya
AU - Nakahara, Makiko
AU - Tourtas, Theofilos
AU - Schlötzer-Schrehardt, Ursula
AU - Kruse, Friedrich
AU - Koizumi, Noriko
N1 - Funding Information:
The authors thank Seiichiro Sugita (Sugita Eye Hospital) for providing research materials, and Emi Ueda and Kyoko Watanabe for technical assistance. Supported by the Program for the Strategic Research Foundation at Private Universities from MEXT (NK, NO), National Institutes of Health grant EY26490 (MPF) and the Robert Waller Career Development Award (KHB).
Publisher Copyright:
© 2019 The Authors.
PY - 2019/2
Y1 - 2019/2
N2 - PURPOSE. CTG trinucleotide repeat (TNR) expansion is frequently found in transcription factor 4 (TCF4) in Fuchs’ endothelial corneal dystrophy (FECD), though the effect of TNR expansion on FECD pathophysiology remains unclear. The purpose of this study was to evaluate the effect of TNR expansion on TCF4 expression in corneal endothelium of patients with FECD. METHODS. Peripheral blood DNA and Descemet membrane with corneal endothelium were obtained from 203 German patients with FECD. The CTG TNR repeat length in TCF4 was determined by short tandem repeat (STR) assays and Southern blotting using genomic DNA. Genotyping of rs613872 in TCF4 was performed by PCR. TCF4 mRNA levels in corneal endothelium were evaluated by quantitative PCR using three different probes. Control corneal endothelial samples were obtained from 35 non-FECD subjects. RESULTS. The STR assay and Southern blotting showed that 162 of the 203 patients with FECD (80%) harbored CTG trinucleotide repeat lengths larger than 50. Quantitative PCR using all three probes demonstrated that TCF4 mRNA is significantly upregulated in the corneal endothelium of patients with FECD, regardless of the presence of TNR expansion. However, the length of the TNR tended to show a positive correlation with TCF4 expression level. No correlation was shown between the genotype of TCF4 SNP, rs613872, and the level of TCF4 expression. CONCLUSIONS. Our findings showed that TCF4 mRNA is upregulated in the corneal endothelium of patients with FECD. Further studies on the effects of TCF4 upregulation on corneal endothelial cell function will aid in understanding the pathophysiology of FECD.
AB - PURPOSE. CTG trinucleotide repeat (TNR) expansion is frequently found in transcription factor 4 (TCF4) in Fuchs’ endothelial corneal dystrophy (FECD), though the effect of TNR expansion on FECD pathophysiology remains unclear. The purpose of this study was to evaluate the effect of TNR expansion on TCF4 expression in corneal endothelium of patients with FECD. METHODS. Peripheral blood DNA and Descemet membrane with corneal endothelium were obtained from 203 German patients with FECD. The CTG TNR repeat length in TCF4 was determined by short tandem repeat (STR) assays and Southern blotting using genomic DNA. Genotyping of rs613872 in TCF4 was performed by PCR. TCF4 mRNA levels in corneal endothelium were evaluated by quantitative PCR using three different probes. Control corneal endothelial samples were obtained from 35 non-FECD subjects. RESULTS. The STR assay and Southern blotting showed that 162 of the 203 patients with FECD (80%) harbored CTG trinucleotide repeat lengths larger than 50. Quantitative PCR using all three probes demonstrated that TCF4 mRNA is significantly upregulated in the corneal endothelium of patients with FECD, regardless of the presence of TNR expansion. However, the length of the TNR tended to show a positive correlation with TCF4 expression level. No correlation was shown between the genotype of TCF4 SNP, rs613872, and the level of TCF4 expression. CONCLUSIONS. Our findings showed that TCF4 mRNA is upregulated in the corneal endothelium of patients with FECD. Further studies on the effects of TCF4 upregulation on corneal endothelial cell function will aid in understanding the pathophysiology of FECD.
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U2 - 10.1167/iovs.18-25760
DO - 10.1167/iovs.18-25760
M3 - Article
C2 - 30811544
AN - SCOPUS:85062425533
VL - 60
SP - 779
EP - 786
JO - Investigative Ophthalmology
JF - Investigative Ophthalmology
SN - 0146-0404
IS - 2
ER -