Chukai Huang

Interactive Effects of ATOH7 and RFTN1 in Association with Adult-Onset Primary Open-Angle Glaucoma

PURPOSE Genome-wide association studies have shown association of the atonal homolog 7 (ATOH7) and raftlin lipid raft linker 1 (RFTN1) genes with glaucoma-related optic disc parameters. ATOH7 and RFTN1 sequence variations were investigated in patients with primary open-angle glaucoma (POAG) and their relationships with vertical cup-to-disc ratio (VCDR) and central corneal thickness (CCT) were determined. METHODS In 289 unrelated controls and 142 patients with adult-onset POAG, including 117 with high-tension glaucoma (HTG) and 25 with normal-tension glaucoma (NTG), the single exon of ATOH7 was sequenced by direct sequencing. Additional single-nucleotide polymorphisms (SNP) at upstream ATOH7 (rs1900004 and rs3858145) and an RFTN1 SNP (rs690037) were genotyped. Quantitative trait and disease associations were analyzed by linear and logistic regression respectively, controlling for sex and age. RESULTS ATOH7 rs61854782 was associated with VCDR (P = 0.004) in controls and RFTN1 rs690037 was associated with CCT in combined POAG (HTG+NTG; P = 0.026). No coding mutation was detected in POAG, and no SNP was associated with POAG (P between 0.441 and 0.996). However, ATOH7 rs3858145 showed significant interaction with RFTN1 rs690037 in NTG and combined POAG (P = 0.026 and 0.013 respectively). ATOH7 rs3858145 GG combined with RFTN1 rs690037 TT conferred risk for glaucoma in HTG, NTG, and combined POAG (odds ratio = 2.11, 8.44, and 2.69, respectively). CONCLUSIONS Coding mutations of ATOH7 were unlikely to be involved in POAG. But combination of ATOH7 and RFTN1 SNPs increased risk to POAG, indicating their diversified effects in the complex genetics of glaucoma.

Association analysis of cigarette smoking with onset of primary open-angle glaucoma and glaucoma-related biometric parameters

BackgroundTo date, studies on the role played by cigarette smoking in primary open-angle glaucoma (POAG) remains controversial. The current study evaluated cigarette smoking as a risk factor of POAG and its relationships with vertical cup-to-disc ratio (VCDR), central corneal thickness (CCT) and intraocular pressure (IOP) in a Chinese cohort.MethodsIn a total of 248 unrelated individuals including 30 juvenile-onset POAG (JOAG), 92 adult-onset POAG (AOAG) and 126 sex-matched senile cataract controls, underwent comprehensive ophthalmic examination. Their smoking was obtained and documented by questionnaire. Association of cigarette smoking with POAG was performed using logistic regression controlled for age and sex. Effects of cigarette smoking on VCDR, IOP and CCT were analyzed with multiple linear regression.ResultsIn either JOAG or AOAG, no association of cigarette smoking was found with disease onset (P = 0.692 and 0.925 respectively). In controls and JOAG, no significant effects of smoking were found on VCDR, IOP or CCT (all P > 0.05). Smoking was found to be correlated with decreased CCT in AOAG and combined POAG (JOAG + AOAG) (P = 0.009 and 0.003), but no association with VCDR or IOP was observed (P > 0.05).ConclusionsAlthough cigarette smoking was not found to be risk factor for onset of POAG, it was correlated with CCT in AOAG, and thus might still play a role in the disease course, especially for AOAG.

Corneal Biomechanics, Refractive Error, and Axial Length in Chinese Primary School Children

PURPOSE Low corneal hysteresis is associated with longer axial length in Chinese secondary school children. The authors sought to explore this association in primary school children. METHODS LogMAR presenting visual acuity, cycloplegic refractive error, ocular biometry, central corneal thickness (CCT), and corneal hysteresis (CH) was assessed for children in grades 1 to 3 at an academically competitive urban school in Shantou, China. RESULTS Among 872 eligible children (mean age, 8.6 ± 2.1 years), 651 (74.7%) completed the examination. Among 1299 examined eyes, 111 (8.5%) had uncorrected vision ≤6/12. Mean spherical equivalent refractive error for all eyes was +0.26 ± 1.41 D, and axial length (AL) was 22.7 ± 0.90 mm. CH for the lowest (mean AL, 21.7 ± 0.39 mm), two middle (mean AL, 22.4 ± 0.15 and 22.9 ± 0.15 mm), and highest quartiles (mean AL, 23.7 ± 0.74 mm) of AL were 10.6 ± 2.1 mm Hg, 10.4 ± 2.1 mm Hg, 10.3 ± 2.3 mm Hg, and 10.2 ± 2.3 mm Hg respectively (age- and gender-adjusted Pearsons correlation coefficient r = -0.052; P = 0.001). In generalized estimating equation models adjusting for age, gender, and CCT, lower CH was significantly associated with longer AL (P < 0.001) and more myopic refractive error (P = 0.001). CONCLUSIONS CH measurement is practical in young children because this is when myopia undergoes its most rapid progression. Prospective follow-up of this cohort at high risk for myopia is under way to determine whether low CH is predictive, or a consequence, of long AL.

Corneal Hysteresis Is Correlated with Reduction in Axial Length After Trabeculectomy

Background: We sought to determine whether corneal biomechanical parameters are predictive of reduction in axial length after anti-metabolite trabeculectomy. Methods: Chinese subjects undergoing trabeculectomy with mitomycin C by a single experienced surgeon underwent the following measurements: Corneal hysteresis (CH, Ocular Response Analyzer, Reichert Ophthalmic Instruments), Goldmann intra-ocular pressure (IOP), central corneal thickness (CCT) and axial length (AL, IOLMaster, Carl Zeiss Meditec, Dublin, CA) were measured pre-operatively, and AL, CH and IOP were measured 1 day and 1 week post-operatively. Results: Mean age of 31 subjects was 52.0 ± 15.2 years, and 15 (48.4%) were female. The mean pre-operative IOP of 21.4 ± 9.3 mmHg was reduced to 8.2 ± 4.6 mmHg 1 day and 11.0 ± 4.4 mmHg 1 week post-operatively (p < 0.001). AL declined from 22.99 ± 0.90 to 22.76 ± 0.87 mm at 1 day and 22.74 ± 0.9 mm at 1 week; 30/31 (%) eyes had a decline in AL (p < 0.001, sign test). In multivariate linear regression models including post-operative data from 1 day and 1 week, greater decline in Goldmann IOP (p < 0.0001, greater pre-op axial length (p < 0.001) and lower pre-operative CH (p = 0.03), were all associated with greater decline in post-operative axial length. Conclusions: Eyes with lesser ability of the ocular coat to absorb energy (lower CH) had significantly greater decrease in axial length after trabeculectomy-induced IOP-lowering.

Determinants of postoperative corneal edema and impact on goldmann intraocular pressure.

Purpose: Recent studies report that increased corneal edema because of contact lens wear under closed lids is associated with elevated Goldmann intraocular pressure (GAT IOP). We sought to assess whether the impact of postoperative corneal edema on GAT IOP would be similar and to determine the differential effect of different amounts of edema. Methods: The setting is a tertiary level cataract clinic in Shantou, China. Pre- and postoperative (day 1) GAT IOP, central corneal thickness (CCT), corneal hysteresis, corneal resistance factor, and radius of corneal curvature were measured for consecutive patients undergoing phacoemulsification surgery by 2 experienced surgeons. Corneal edema was calculated as the percentage increase in CCT. Results: Among 136 subjects (mean age, 62.5 ± 15.4 years; 53.7% women), the mean increase in CCT was 10.3% postoperatively. Greater corneal edema was associated with lower GAT IOP in unadjusted analyses (P < 0.03) and in linear regression models (P < 0.01). In the model, higher corneal resistance factor (P < 0.001), lower corneal hysteresis (P < 0.001), and steeper radius of corneal curvature (P < 0.001) were associated with higher GAT IOP. Among subjects with edema < the median, edema was associated with lower GAT IOP (P = 0.004), whereas among those with edema ≥ the median, edema was not associated with GAT IOP. An increase in CCT of 7% was associated with an 8 mm Hg underestimation of GAT IOP in our models. Conclusions: The effect of postoperative edema on GAT IOP seems to be the opposite of contact lens-induced edema. The magnitude of the effect is potentially relevant to patient management.

Mutations of RagA GTPase in mTORC1 Pathway Are Associated with Autosomal Dominant Cataracts.

Cataracts are a significant public health problem with no proven methods for prevention. Discovery of novel disease mechanisms to delineate new therapeutic targets is of importance in cataract prevention and therapy. Herein, we report that mutations in the RagA GTPase (RRAGA), a key regulator of the mechanistic rapamycin complex 1 (mTORC1), are associated with autosomal dominant cataracts. We performed whole exome sequencing in a family with autosomal dominant juvenile-onset cataracts, and identified a novel p.Leu60Arg mutation in RRAGA that co-segregated with the disease, after filtering against the dbSNP database, and at least 123,000 control chromosomes from public and in-house exome databases. In a follow-up direct screening of RRAGA in another 22 families and 142 unrelated patients with congenital or juvenile-onset cataracts, RRAGA was found to be mutated in two unrelated patients (p.Leu60Arg and c.-16G>A respectively). Functional studies in human lens epithelial cells revealed that the RRAGA mutations exerted deleterious effects on mTORC1 signaling, including increased relocation of RRAGA to the lysosomes, up-regulated mTORC1 phosphorylation, down-regulated autophagy, altered cell growth or compromised promoter activity. These data indicate that the RRAGA mutations, associated with autosomal dominant cataracts, play a role in the disease by acting through disruption of mTORC1 signaling.

Detection of mutations in MYOC , OPTN , NTF4 , WDR36 and CYP1B1 in Chinese juvenile onset open-angle glaucoma using exome sequencing

Juvenile onset open-angle glaucoma (JOAG) affects patients before 40 years of age, causing high intraocular pressure and severe optic nerve damage. To expand the mutation spectrum of the causative genes in JOAG, with a view to identify novel disease-causing mutations, we investigated MYOC, OPTN, NTF4, WDR36 and CYP1B1 in a cohort of 67 unrelated Chinese JOAG patients. Whole exome sequencing was used to identify possible pathogenic mutations, which were further excluded in normal controls. After sequencing and the use of a database pipeline, as well as predictive assessment filtering, we identified a total of six mutations in three genes, MYOC, OPTN and CYP1B1. Among them, 2 heterozygous mutations in MYOC (c. 1109C > T, p. (P370L); c. 1150G > C, p. (D384H)), 2 heterozygous mutations in OPTN (c. 985A > G, p.(R329G); c. 1481T > G, p. (L494W)) and 2 homozygous mutations in CYP1B1 (c. 1412T > G, p.(I471S); c. 1169G > A, p.(R390H)) were identified as potentially causative mutations. No mutation was detected in NTF4 or WDR36. Our results enrich the mutation spectra and frequencies of MYOC, OPTN and CYP1B1 in JOAG among the Chinese population. Further studies are needed to address the pathogenicity of each of the mutations detected in this study.

Associated factors of acute primary angle closure glaucoma in a sub-group of Chinese people: comparison between attack eyes and normal controls

Acute primary angle closure glaucoma (APACG) is an ophthalmic emergency that may lead to irreversible blindness. Although efforts were made to control intraocular pressure, disease progression still existed. Anterior segment optical coherence tomography (AS OCT) may provide a new insight into mechanism of APACG. In order to seek out associated factors by AS OCT, We compared anterior segment parameters between 74 APACG and 48 normal eyes. Analysis of variance, principle component analysis and logistic regression were used. APACG group had more women (75.7% vs 47.9%, p = 0.002), smaller anterior chamber (anterior chamber depth, ACD; anterior chamber area, ACA; all p = 0.001), narrower angle (AOD500, AOD750, angle opening distance at 500um and 750um; TISA500, TISA750, trabecular-iris space area at 500um and 750um; ARA500, ARA750, angle recess area at 500um and 750um; all p < 0.001), thinner iris (iris thickness at 750um, IT750; maximum of iris thickness, ITMAX; all p < 0.001), larger iris area (IA, p < 0.001) and lens vault (LV, p = 0.005). Principle component extracted were AOD500, AOD750, TISA500, TISA750, ARA500, ARA750, IA, PD (pupillary diameter), ACD, ACA and IT750. After adjusting for confounding factors, AOD750 (b = 12.40 ± 2.56, p < 0.001), IT750 (b = 10.50 ± 3.45, p = 0.002) and IA (b = −1.56 ± 0.77, p = 0.044) were significantly associated with occurrence of APACG.

Reliability of Tonolab measurements in rats

AIM To assess the repeatability and reproducibility of Tonolab tonometer in rats with high intraocular pressure (IOP) and evaluate its ability to detect IOP changes in rats with general anaesthesia. METHODS Left eyes of adult Fischer rats (F344) were photocoagulated by 532 nm diode laser to induce high IOP. Hypertensive eyes of 30 conscious rats were randomly chosen to measure IOP on a single occasion. Two observers independently and alternately undertook IOP measurements consecutively for three times using the same Tonolab tonometer blind to the other observers IOP measurements. The within subject standard deviation (Sw), coefficient of variation (CVw) (100×Sw/overall mean), and intraclass correlation coefficient (ICC) were calculated to evaluate intra-observer repeatability. Inter-observer difference was analysed by using 95% limits of agreement described by Bland-Altman and paired sample t-test. Also, another 13 normal F344 rats were intraperitoneally administrated with ketamine/xylazine or chloral hydrate, and IOPs of both eyes were measured by a single operator once every 5min until animals came to conscious. IOPs at various time points were compared by using one-way ANOVAs. RESULTS Mean IOP was 35.58 mm Hg (range 17.33 to 65.33 mm Hg). For intraobserver repeatability, the Sw, CVw and ICC of high IOP for two observers were 5.20 mm Hg/3.41 mm Hg, 9.98%/8.08% and 0.820/0.928 respectively. The inter-observer difference was 14.76%±19.76% of the mean IOP of two observers, with a 95% limits of agreement -23.97% to 53.50%, and the difference between mean IOP of these two observers was statistically significant (P=0.001). IOPs dropped slightly during the first 15min post-aneathesia, with a IOP change between 0.17 and 1.17 mm Hg. IOPs changed from basline of 11.75±2.05 mm Hg (n=12) to 8.75±1.06 mm Hg 20min post-anesthesia (P=0.001), and this hypotensive condition persisted until 80min post-anesthesia. CONCLUSION In this sample of hypertensive rats, Tonolab measurements demonstrated high levels of intraobserver repeatability, however, its interobserver reproducibility was poor. Longitudinal changes of IOP caused by genral anaesthesia can be sensitively detected by Tonolab. So we suggested that measurements of IOP using Tonolab are best measured by a single observer, and it could be included in experimental glaucoma.