Figure 1.
Pedigrees of 3 families with PACD.
A. Family 1 (abbreviated). B. Family 2. C. Family 3. Families 1[1] and 3[7] have been reported previously. Asterisks indicate individuals in whom genetic testing was performed. Filled symbols represent affected individuals; open symbols represent unaffected individuals; ? represents individuals of undetermined affected status; arrowhead indicates proband.
Table 1.
Quantitative PCR oligonucleotides.
Figure 2.
Clinical findings from Family 2.
A. Slit lamp photomicrograph demonstrating peripheral corneal opacification in individual IV-5. B–C. Slit lamp photomicrograph of central and peripheral corneal opacification in individuals III-6 (B) and III-2 (C). D. Corneal topographic imaging demonstrates significant flattening of the corneal curvature, with a steep K value of 38.79 D, in individual III-2.
Figure 3.
Results of genomic CNV analysis using the CytoScan® HD Array in 3 families with PACD.
A schematic of the genes in the region of chromosome 12q21.33 is shown at the top. In Family 1, an unaffected individual (IV-17) without a deletion in the region of chromosome 12q21.33 is shown above an affected individual (III-4) who demonstrates a 701 Kb heterozygous deletion involving CCER1, KERA, LUM, DCN, and EPYC. In Family 2, a 1.318 Mb heterozygous deletion involving CCER1, KERA, LUM, DCN, and EPYC was identified in an affected individual (IV-2). In Family 3, a 701 Kb heterozygous deletion in chromosome 12q21.33 with identical boundaries to that identified in Family 1 was identified in an affected individual (III-4).
Figure 4.
Quantitative PCR results for EPYC and DCN for 3 families with PACD.
The cutoff between 1 and 2 copies is represented by a horizontal red line at 1.414, the geometric mean between 1 and 2. Results are grouped by affected status and arranged in order of location on pedigree. Data are represented as the mean± SEM. A = affected; U = unaffected; *Genomic CNV analysis also performed.
Figure 5.
Expression of the SLRP gene cluster and CCER1 in the corneal stroma by qPCR.
A. The transcript levels for the keratocyte markers CD34 and VIM were significantly higher in the corneal stroma than in the corneal endothelium. Statistical analysis was performed using a two-tailed unpaired t-test. B. Expression of LUM and KERA was significantly higher than DCN, which in turn was higher than the undetectable expression levels of EPYC and CCER1. Statistical analyses were performed using one-way ANOVA and Bonferroni's Multiple Comparison Test (*p≤0.05; **p≤0.01; ***p≤0.001 (error bars = SEM)). Non-significant results are not represented.