Next-generation sequencing-based clinical diagnosis of choroideremia and comprehensive mutational and clinical analyses.

BACKGROUND: To report the clinical and genetic findings from seven Chinese patients with choroideremia. METHODS: Five hundred seventy-eight patients with a clinically suspected diagnosis of retinitis pigmentosa (RP) underwent comprehensive ophthalmic examinations. Next-generation sequencing (NGS) was performed on samples from all patients. Detailed clinical characteristics of the patients with choroideremia identified in this study were assessed using multimodal imaging. RESULTS: Seven patients with choroideremia were identified, and six novel variants in CHM (c.1960 T > C p.Ter654Gln, c.1257del p.Ile420*fs1, c.1103_1121delATGGCAACACTCCATTTTT p.Tyr368Cysfs35, c.1414-2A > T, and c.1213C > T p.Gln405Ter, c.117-1G > A) were revealed. All variants were deleterious mutations: two were frameshifts, two were nonsense mutations, two were splicing mutations, and one was a readthrough mutation. The clinical phenotypes of these patients were markedly heterogeneous, and they shared many common clinical features with RP, including night blindness, constriction of the visual field and gradually reduced visual acuity. However, patients with choroideremia showed pigment hypertrophy and clumping, and chorioretinal atrophy, and a majority of patients with choroideremia presented with retinal tubulations in the outer layer of the retina. CONCLUSIONS: We provide a detailed description of the genotypes and phenotypes of seven patients with choroideremia who were accurately diagnosed using NGS. These findings provide a better understanding of the genetics and phenotypes of choroideremia.

Clinical and Genetic Characteristics of Chinese Patients with Occult Macular Dystrophy.

Purpose: To investigate the clinical and genetic characteristics of occult macular dystrophy (OMD) based on a Chinese patient cohort. Methods: Fifteen Chinese OMD patients from nine unrelated families underwent genetic testing, and all of them harbored a pathogenic RP1L1 variant. Comprehensive ophthalmic examinations were performed in nine probands, including spectral-domain optical coherence tomography (SD-OCT), near-infrared reflectance (NIR), fundus autofluorescence (AF), and multifocal electroretinography. Results: The RP1L1 variants p.R45W and p.S1199C were identified in 13 patients and two patients, respectively, and one was a de novo mutation. Among the nine probands, the median ages at onset and examination were 25.0 years (range, 6-51 years) and 27.0 years (range, 14-55 years), respectively. The median decimal visual acuity was 0.20 (range, 0.04-0.5). Foveal photoreceptor thickness and visual acuity showed a significant correlation (r = 0.591; P = 0.01). All eyes presented with an absent interdigitation zone and blurred ellipsoid zone of photoreceptors when examined by SD-OCT. In addition, central round lesions with low NIR reflectance were observed in 66.7% (12/18) of eyes by NIR reflectance imaging, corresponding to the regions with abnormal photoreceptor microstructures observed by SD-OCT. Of the 18 eyes, only four eyes showed ring-like faint hyperfluorescence around the macula by AF. Conclusions: To the best of our knowledge, this is the largest study in a cohort of Chinese OMD patients with RP1L1 mutations. Our findings revealed that the two recurrent RP1L1 variants are related to OMD in the Chinese population. Furthermore, multimodal imaging combined with genetic testing is valuable for diagnosing and monitoring OMD progression.

The Precise Diagnosis of Wolfram Syndrome Type 1 Based on Next-Generation Sequencing.

Purpose: To explore a method for the early, rapid and accurate diagnosis of Wolfram syndrome 1 (WS1) and further enrich the spectrum of WFS1 mutations in the Chinese population. Methods: We analyzed 279 patients with unexplained optic atrophy using next-generation sequencing. All patients underwent detailed clinical evaluations. Furthermore, Sanger sequencing and cosegregation analyses were performed within families. Results: Five patients with WS1 were identified in four unrelated families, and their clinical features were reviewed in detail. Seven variants of WFS1 were detected, including three reported variants (p.G674R, p.Tyr508Cysfs*34, and p.G702D) and four novel variants (p.W540G, p.K634*, p.F770C, and p.Q584P). Furthermore, the variant p.G674R was recurrent. Conclusion: Considering that WS1 is a rare progressive neurodegenerative disease, early diagnosis is beneficial to the systematic evaluation, monitoring and management of complications to improve patient quality of life and delay the progression of the disease. In the future, precise diagnosis on the basis of clinical manifestation and genetic testing will become the gold standard for the diagnosis of hereditary eye diseases and syndromes. Finally, our results further increase the spectrum of WFS1 mutations by adding four novel variants to the limited data available in the Chinese population.

Next-Generation Sequencing-Aided Rapid Molecular Diagnosis of Occult Macular Dystrophy in a Chinese Family.

Purpose: To show early, rapid and accurate molecular diagnosis of occult macular dystrophy (OMD) in a four-generation Chinese family with inherited macular dystrophy. Methods: In the current study, we comprehensively screened 130 genes involved in common inherited non-syndromic eye diseases with next-generation sequencing-based target capture sequencing of the proband of a four-generation Chinese family that has suffered from maculopathy without a definitive diagnosis for over 10 years. Variants were filtered and analyzed to identify possible disease-causing variants before validation by Sanger sequencing. Results: Two heterozygous mutations-RP1L1 c.133 C > T (p.Arg45Trp), which is a hot spot for OMD, and ABCA4 c.6119 G > A (p.Arg2040Gln), which was identified in Stargardt's disease were found in three patients, but neither of the mutations was found in the unaffected individuals in the same family, who are phenotypically normal or in the normal control volunteers. Conclusion: These results cannot only confirm the diagnosis of OMD in the proband, but also provide presymptomatic diagnosis of the proband's children before the onset of visual acuity impairment and guidance regarding the prognosis and management of these patients. Heterozygous mutations of RP1L1 c.133 C > T (p.Arg45Trp) and ABCA4 c.6119 G > A (p.Arg2040Gln) are likely responsible for OMD. Our results further extend our current understanding of the genetic basis of OMD, and emphasize the importance of molecular diagnosis and genetic counseling for OMD.

[Etiologic analysis of 487 cases of ophthalmoplegia].

OBJECTIVE: To study the etiology of ophthalmoplegia cases. METHODS: A retrospective case series study. We summarized and analyzed etiological diagnosis of 487 ophthalmoplegia patients from January 2005 to September 2010 in Beijing Tongren Hospital of Capital Medical University and Beijing Tongren Eye Center. Clinical data included the case history, clinical manifestations, and results of examinations of neurology, ophthalmology, endocrinology and iconography. The analysis of variance (ANOVA) and Chi-Square test were used in our study. RESULTS: Nineteen different kinds of causes were identified. In terms of age onset, microvascular ischemic (MVI) patients were the oldest (60.38 +/- 11.16) in all groups. It significantly distinguished from myasthenia gravis (MG) and local non-specific inflammation (F = 24.46, P = 0.000). From the view of ophthalmoplegia characters, bilateral asymmetry ophthalmoplegia was the character of MG. We also found that all MVI patients had lesions in unilateral single ocular movement nerve. Unilateral multiple nerves or muscles lesions were the main feature of local non-specific inflammation. In addition, from the view of concomitant symptoms, local aching was very frequent in local non-specific inflammation (all 60 cases) and MVI (44 cases) patients (Chi2 = 36.346, P = 0.000). The mild pupil changing could be found in about one half patients of the two diseases (Chi2 = 0.026, P = 0.875). CONCLUSIONS: The causes of ophthalmoplegia are very complicate. MG, MVI and local non-specific inflammation are the most frequent causes. In more than half of patients, the lesions are located in neurological system, about one third located in neuromuscular junction and the least in the muscles.

[Clinical characteristics of acute ischemic retinopathy and nonarteritic anterior ischemic optic neuropathy].

OBJECTIVE: To compare the vascular risk factors between acute ischemic retinopathy (AIR) and nonarteritic anterior ischemic optic neuropathy (NAION). METHODS: Single center retrospective study. Patients diagnosed of AIR and NAION from Jan 2008 to Jan 2009 in Beijing Tongren hospital were enrolled in this study. The demographical data, the results of blood tests, cardiac examinations, carotid vascular ultrasound and brain MRI were analyzed by Chi-square test. RESULTS: Ninety-one AIR patients and 143 NAION patients were enrolled. The mean age of the NAION patients (51.4 ± 8.9) was lower than that of the AIR group (55.4 ± 15.8) (t = 2.514, P = 0.013). There was no significant difference in the gender, smoking status, hypertension and fasting blood glucose levels between these two groups (χ(2) = 0.023, 2.727, 0.217, 0.012; all P > 0.05). The percentage of hypercholesterolemia (total cholesterol > 5.17 mmol/L) was 46.2% (66/143) in the NAION group and 26.4% (24/91) in the AIR group (χ(2) = 9.193, P = 0.002). Prevalence of carotid artery disease and ischemic heart disease was 13.2% (12/91) in the AIR group and 4.9% (7/143) in the NAION group (χ(2) = 5.125, P = 0.024). Carotid artery stenosis was present in 34.5% (30/87) patients in the AIR group, whereas none in the NAION group (χ(2) = 44.370, P = 0.000). Among 57 AIR patients and 127 NAION patients who finished brain MRI, the percentage of cerebral infarction was 47.4% (27/57) in the AIR group and 0.8% (1/127) in the NAION group (χ(2) = 66.210, P = 0.000). Cerebral white matter hyperintensities (WMH) were present in 66.9% (85/127) of the NAION group and 36.8% (21/57) in the AIR group (χ(2) = 66.46, P = 0.000). CONCLUSIONS: The mean age of NAION patients was lower than that of the AIR patients. Carotid artery stenosis and stroke, which might indicate large-middle vessel disease, are common in ischemic retinopathy. WMH with small vascular atherosclerosis, possibly caused by hypercholesterolemia, is more common in the NAION group.

[Clinical characteristics of non-arteritic anterior ischemic optic neuropathy].

OBJECTIVE: To evaluate the demographic, risk factors and clinical characteristics of non-arteritic anterior ischemic optic neuropathy (NAION) and provide clinical guidance for this blindness disease. METHODS: Retrospective study was used to investigate the data, which consists of 96 NAION consecutive patients in neurology department of our hospital from 2005 to 2008. RESULTS: The average age of NAION patients is 50.90 +/- 8.88 years (range, 34 - 78 years) and the median is 50 y. About 68.8% of patients are male. The prevalence of hypertension, diabetes mellitus and abnormal lipid level is 34.4% (33/96), 29.2% (21/96) and 59.4% (57/93) respectively. The predominant patterns of visual field loss are altitudinal and accurate defects. Visual acuity was improved (change of >/= 3 lines) in 44.4% of patients. CONCLUSIONS: The age of onset in NAION is relatively early in life, compared with cerebral vascular disease, and male seems to have a high incidence. The most common risk factor is metabolic syndrome. Subcortical and periventricular white matter lesions and myocardial ischemia might associate with NAION. Visual acuity showed improvement in some patients, but the long outcome is poor.

[Clinical features and therapy of subjective benign paroxysmal positional vertigo].

OBJECTIVE: To evaluate the clinical features and therapy of subjective benign paroxysmal positional vertigo (S-BPPV). METHODS: By retrospectively analyzing the results of clinical features and therapy in 12 patients with S-BPPV from January 2003 to September 2006, the results were compared with 24 patients with objective benign paroxysmal positional vertigo (O-BPPV) of posterior semicircular canal. RESULTS: S-BPPV patients suffered from attack of transient vertigo with sudden onset triggered by head motion but no concomitant nystagmus in Dix-Hallpike test. The latency and duration of vertigo attack were (4.42 +/- 2.02) s and (8.67 +/- 4.31) s in S-BPPV, (3.2 +/- 1.18) s and (14.75 +/- 4.97) s in O-BPPV of posterior semicircular canal. The differences between the two groups were all significant (t = 2.30, P < 0.05 and t = 3.61, P < 0.01). The symptoms disappeared in 11 patients after a single therapy of particular repositioning maneuver and 1 patient after 2 times therapy in S-BPPV. The one-stage success rate was 91.7% in S-BPPV and 79.2% in O-BPPV of posterior semicircular canal, but the difference between these two groups was not significant. The number of circulation therapy in first management was (1.75 +/- 1.08) times in S-BPPV and (3.38 +/- 1.06) times in O-BPPV of posterior semicircular canal, while the difference was significant (t = 4.32, P < 0.01). There were 2 patients recurred during follow-up in S-BPPV and 7 patients in O-BPPV of posterior semicircular canal, but the difference wasn't significant. CONCLUSIONS: Longer latency, shorter duration and need less circulation therapy are achieved in S-BPPV compared with O-BPPV of posterior semicircular canal, which indicate that the effectiveness of S-BPPV seems to be more favorable than that of O-BPPV of posterior semicircular canal.