Ana Laura Canedo

Genetic association of COL5A1 variants in keratoconus patients suggests a complex connection between corneal thinning and keratoconus

PURPOSE Single nucleotide polymorphisms (SNPs) located near or within the COL5A1 gene, at 9q34.2-q34.3 chromosomal region have been reported in association with central corneal thickness (CCT). Using family linkage analysis, we identified a keratoconus susceptibility locus at 9q34. These findings led us to perform an association study between COL5A1 variation and keratoconus susceptibility. METHODS A Caucasian case-control cohort of 222 keratoconus patients and 3324 controls was selected as the discovery panel. An independent case-control panel of 304 cases and 518 controls and a family panel of 186 subjects were replicated for genotyping and association. Forty-four SNPs (21 for discovery and 23 for fine-mapping) spanning 300 kilobases in and around COL5A1 were genotyped and tested for genetic association. Logistic regression models implemented in PLINK were used to test for association in case controls. Generalized estimating equation models accounting for familial correlations implemented in genome-wide interaction analyses with family data were used for association testing in families. RESULTS Two CCT associated SNPs (rs1536482 and rs7044529 near and within COL5A1) were identified in the keratoconus discovery cohort (P values of 6.5 × 10(-3) and 7.4 × 10(-3)). SNP rs1536482 was replicated in the second case-control sample (P = 0.02), and SNP rs7044529 was replicated in a keratoconus family panel (P = 0.03). Meta P values of rs1536482 and rs7044529 in the keratoconus cohorts were 1.5 × 10(-4) (odds ratio [OR] = 1.30) and 2.9 × 10(-3) (OR = 1.39). After Bonferroni correction, the association of SNP rs1536482 remained significant (P = 6.5 × 10(-3)). CONCLUSIONS SNPs in the COL5A1 region, which regulate normal variation in CCT, may play a role in the thinning associated with keratoconus.

C.57 C > T Mutation in MIR 184 is Responsible for Congenital Cataracts and Corneal Abnormalities in a Five-generation Family from Galicia, Spain

Abstract A c.57 C > T mutation in the seed region of MIR184 located at the 15q25.1 chromosomal region has been independently associated with autosomal dominant keratoconus with early-onset anterior polar cataract in the Northern Irish family and with autosomal dominant EDICT (Endothelial Dystrophy, Iris hypoplasia, Congenital cataracts, and stromal Thinning) syndrome. In this study we report a five-generation family originating in Galicia, Spain with early onset cataracts and variable corneal abnormalities which include non-ectatic corneal thinning and severe early-onset keratoconus. We identified a heterozygous c.57 C > T mutation in miR-184 in the proband and two additional affected relatives on the maternal side. This finding represents a third independent occurrence of this mutation in familiar ocular disease thus strengthening the link between miR-184 abnormalities and inherited eye defects.

Assessing ectasia susceptibility prior to LASIK: the role of age and residual stromal bed (RSB) in conjunction to Belin-Ambrósio deviation index (BAD-D)

Purpose: To compare the ability to detect preoperative ectasia risk among LASIK candidates using classic ERSS (Ectasia Risk Score System) and Pentacam Belin-Ambrosio deviation index (BAD-D), and to test the benefit of a combined approach including BAD-D and clinical data. Methods: A retrospective nonrandomized study involved preoperative LASIK data from 23 post-LASIK ectasia cases and 266 stable-LASIK (follow up > 12 months). Preoperative clinical and Pentacam (Oculus; Wetzlar, Germany) data were obtained from all cases. Mann-Whitneys test was performed to assess differences between groups. Stepwise logistic regression was used for combining parameters.The areas under the Receiver Operating Characteristic (ROC) curves (AUC) were calculated for all parameters and combinations, with pairwise comparisons of AUC (DeLongs method). Results: Statistically significant differences were found for age, residual stromal bed (RSB), central corneal thickness and BAD-D (p 0.05). ERSS was 3 or more on 12/23 eyes from the ectasia group (sensitivity = 52.17%) and 48/266 eyes from the stable LASIK group (18% false positive). BAD-D had AUC of 0.931 (95% CI: 0.895 to 0.957), with cut-off of 1.29 (sensitivity = 87%; specificity = 92.1%). Formula combining BAD-D, age and RSB provided 100% sensitivity and 94% specificity, with better AUC (0.989; 95% CI: 0.969 to 0.998) than all individual parameters (p>0.001). Conclusion: BAD-D is more accurate than ERSS. Combining clinical data and BAD-D improved ectasia susceptibility screening. Further validation is necessary. Novel combined functions using other topometric and tomographic parameters should be tested to further enhance accuracy.

Corneal collagen cross-linking for keratoconus and post-LASIK ectasia.

Corneal collagen cross-linking (CXL) is a novel technology that utilizes a combination of riboflavin (vitamin B2) eye drops, to saturate the cornea, and UV-A light which sets off a chemical reaction to shorten the cross links between and within collagen fibers thereby increasing the biomechanical stability of the corneal stroma (1–4). The objective of this procedure is to halt progression of keratoconus, post-LASIK ectasia, and other corneal ectasias, such as pellucid marginal degeneration. In some cases, there has been improvement in visual acuity, reduction of corneal curvature and improvement in quality of life (5). In the 1970’s Siegel discussed cross-linking reactions whereby lysyl oxidase catalyzed the formation of cross-linking aldehydes in collagen and elastin (6–7). Cross-linking is well known in material sciences and dentistry where this enzymatic process increases molecular bonds to increase the mechanical strength of tissue. Cross-linking can be induced enzymatically by means of aldehydes, chemical fixatives or by photosensitizing radiation. Of the three methods the photosensitizing radiation has been shown to be most effective. In corneal clinical practice, CXL utilizes this method where the riboflavin acts as a photosensitizer for the production of free radicals produced by the interaction of the riboflavin and UV-A light. The production of these reactive oxygen species (singlet oxygen) causes the formation of chemical bonds within the corneal stroma resulting in corneal stiffening (3).

Perfil paquimétrico horizontal para a detecção do ceratocone

Purpose: To evaluate the ability of horizontal thickness profile to distinguish keratoconus from normal corneas, and compare the accuracy of these indices with more complex tomographic indices. Methods: In a retrospective study, one eye randomly selected from 225 patients with bilateral keratoconus and 335 patients with normal corneas were included. All patients were examined by a corneal specialist and underwent the examination of corneal tomography, Pentacam HR (Oculus, Wetzlar, Germany). Data of horizontal pachymetric profile passing through the apex of 6 mm was evaluated. With these data, the thickness at the corneal apex (P.apice), the thickness of the thinnest point of the horizontal profile (P.min.H), the pachymetric progression from the thinnest point to the thickest (PPmin-max), the average pachymetric progression in the meridian as reference the average from the normal population (PPmed) and the fitness with a second degree polynomial trendline (R2). An artificial intelligence model was built to combine this data. The performance for distinguishing normal keratoconus was evaluated by ROC curves. Results: All of these indices were statistically different between the two groups (p <0.001). The area under the ROC curve (AUC) thickness at the apex and TP were 0.904 and 0.938, respectively. The tomographic index with higher AUC was BAD-D (0.997). Regarding the horizontal profile, the AUC of PP.min.H and PPmin-max were 0.915 and 0.927, respectively. The fitness to the trendline to the horizontal thickness graph AUC was 0.896. The best performance was obtained with the PPmed (AUC 0.932 sensitivity = 84.4% and specificity of 92.5%). The artificial intelligence model combining all items derived from horizontal profile improved performance (AUC 0.991 Sensitivity = 96% and specificity of 98%). Conclusion: The horizontal thickness profile can detect keratoconus, with a capacity comparable to more complex indices. This type of analysis can provide the basics for new approaches, using data present in simpler devices than the tomographer reducing the cost for the patients.

Differentiation of mild keratoconus from corneal warpage according to topographic inferior steepening based on corneal tomography data

We report two cases of suspicious asymmetric bow tie and inferior steepening on topographic evaluations with reflection (Placido) and projection (Scheimpflug). Rotating Scheimpflug corneal and anterior segment tomography (Oculus Pentacam HR, Wetzlar, Germany)® was performed in the first case, with a maximal keratometric value (Kmax) of 43.2 D and an overall deviation value from the Belin/Ambrósio Enhanced Ectasia Display (BAD-D) of 1.76, which was observed in the study eye (OD). BAD-D was 6.59 in the fellow eye, which had clinical findings that were consistent with keratoconus stage 2. The second case presented with a Kmax of 45.3 D and BAD-D of 0.76 in OD and 1.01 in OS. This patient had discontinued wearing soft contact lens less than 1 day prior to examination. Corneal tomographic data enabled us to distinguish mild or forme fruste keratoconus from contact lens-induced corneal warpage, and similar findings were observed on curvature maps.