先天性无虹膜合并先天性白内障家系致病基因突变分析

目的：对先天性无虹膜合并先天性白内障家系进行PAX6基因突变位点筛查,丰富该致病基因的突变谱。

方法：选取就诊于北京同仁医院眼科门诊的1个先天性无虹膜合并先天性白内障家系和100名健康志愿者,采集外周静脉血,提取基因组DNA,采用直接测序法进行PAX6基因突变位点的筛查。

结果：该家系中先证者和其他患者均表现为无虹膜合并白内障,PAX6基因测序结果显示,该致病基因第11外显子无义突变c.991 C>T,造成PAX6基因编码的蛋白截短(R331X),从而使该蛋白失去功能,且该突变在家系内与疾病表型共分离,不存在于家系内及家系外健康样本的基因中。

结论：PAX6 R331X突变与先天性无虹膜合并先天性白内障的发生有关。     

A novel PAX6 mutation causes congenital aniridia with or without retinal detachment

Background: Aniridia is a rare developmental eye disorder characterized by complete or partial iris hypoplasia often accompanied with other ocular changes that affect the cornea, anterior chamber, lens, retina, and optic nerve. Most cases of aniridia are inherited with an autosomal dominant mode of inheritance caused by PAX6 mutations or deletions. To reveal the underlying genetic defect in a four-generation Iranian family with aniridia, we carried out a genetic screening of PAX6.

Methods: Complete ophthalmic examinations were performed for available affected family members. All PAX6 exons and their flanking regions were sequenced for affected individuals. Candidate variation was screened for segregation in the pedigree by Sanger sequencing. Bioinformatics prediction was done to evaluate the deleterious effects of the mutation on protein product. Real-time PCR was used to investigate the impact of the variant on PAX6 mRNA expression.

Results: All patients were diagnosed with isolated aniridia associated with variable phenotypic features including retinal detachment. A novel heterozygous deletion c.320_348delTGTCCGAGGGGGTCTGTACCAACGATAAC (p.Leu107HisfsX16) on PAX6 gene was detected. Decreased mRNA level of PAX6 in the affected individuals indicated that the mutation caused nonsense-mediated mRNA decay (NMD).

Conclusions: To the best of our knowledge, it is the first report on the genetics of aniridia in Iran. Segregation analysis, bioinformatics prediction and confirmation of NMD, all support the proposition that the novel observed PAX6 mutation is the cause of aniridia in the pedigree. Retinal detachment in some of the affected members, which is a rare reported phenotypic feature of aniridia patients, may be associated with this mutation.     

PAX6基因突变的鉴定与中国家系无虹膜症合并妊娠期糖尿病的产前诊断

目的：探讨1个中国无虹膜症合并妊娠期糖尿病家系的基因缺陷及产前诊断。

方法：收集1个患有无虹膜症合并妊娠期糖尿病的中国家系,从外周血中提取整个家系成员的基因组DNA,通过聚合酶链式反应结合直接测序法,分析人类配对盒基因(PAX6)的编码序列。妊娠18wk时对孕妇进行羊膜穿刺术,并根据突变筛查结果进行遗传学分析。

结果：无虹膜患者在PAX6的第5外显子中存在杂合缺失突变(c.113_129del GGCCGTGCGACATTTCC, p.Arg38ProfsTer12),该患者同时合并妊娠期糖尿病,产前诊断结果提示胎儿具有相同的突变,易患先天性无虹膜症,经产后随访证实。

结论：在中国先天性无虹膜患者中发现了PAX6基因缺失突变,为人类PAX6等位基因变异数据库提供了更多的文献资料,为产前诊断提供了分析依据。     

PAX6 3′ deletion in a family with aniridia

Background: Aniridia is a congenital panocular malformation defined as iris aplasia or hypoplasia. It can be either isolated or be a part of multiple ocular anomalies such as cataracts, glaucoma, corneal pannus, optic nerve hypoplasia, absence of macular reflex or ectopia lentis. In the majority of cases the disease is caused by mutation in the PAX6 gene.

Material and Methods: A Polish family with aniridia was screened for the presence of genomic rearrangements in PAX6, WT1 and the flanking genes by means of multiplex ligation probe amplification (MLPA). MLPA reaction was performed using the P219-B1 PAX6 commercial kit from MRC-Holland. Additionally, the coding sequence of PAX6 gene was sequenced in the proband. Array comparative genomic hybridization analysis was performed using the NimbleGen CGX-12 format.

Results: MLPA examination revealed a heterozygous deletion of approximately 0.6?Mb, downstream of PAX6 gene on chromosome 11. Four genes lie in the deleted region. Bi-directional sequencing of 14 exons of the PAX6 gene did not reveal any causative alteration. Microarray analysis confirmed the deletion and determined its size which ranged from 598.87–651.76 kb.

Conclusions: A small subset of aniridia cases is caused by rearrangements of PAX6 neighboring regions, and the so-called “position effect” is considered to be the underlying pathogenic mechanism. Molecular testing of aniridia patients should include sequencing of the PAX6 gene, followed by screening for larger structural abnormalities located on chromosome 11p13. MLPA can be a useful method in molecular testing of aniridia patients.     

先天性无虹膜家系中发现PAX6基因新的致病突变

目的对中国一先天性无虹膜家系进行PAX6基因突变检测,以确定其突变位点。方法实验研究。收集一先天性无虹膜家系,采集该家系患者及健康成员的外周静脉血,收集100名健康人外周血作为正常对照,采用Sanger测序的方法对PAX6基因的11个外显子（外显子4-14）以及外显子-内含子连接区域进行测序,随后进行家系共分离分析以及正常样本的对照分析。结果该家系8名成员经全面眼科检查,3名确诊为先天性无虹膜,且合并有复杂的眼部表型,包括不同程度的角膜病变、不同类型的白内障、黄斑发育不良、轻度上睑下垂和轻度眼球水平震颤等。在该家系患者中发现一个新杂合突变[c.569_570delinsACGG（p.Ile190Asnfs*18）],该突变可致PAX6基因编码的蛋白截短,该突变符合共分离且在100名正常对照者中未检测到。结论PAX6基因第8外显子上一个新的杂合突变[c.569_570delinsACGG（p.Ile190Asnfs*18）]为本研究中先天性无虹膜家系的致病突变,该突变与先天性无虹膜有关,本研究扩大了PAX6基因的突变频谱。     

先天性无虹膜症家系的基因突变位点研究

目的探讨先天性无虹膜症家系的基因突变位点。方法抽取家系成员的外周血2~5ml,提取DNA;先合成2个多态性微卫星遗传标记(D11S904和D11S935)的引物进行聚合酶链反应(PCR),PCR产物变性后用变性聚丙烯酰胺(PAGE)胶分离,根据带型和家系成员间的关系进行单体型连锁分析,判断家系无虹膜表型是否与PAX6基因相关;PCR扩增PAX6基因的所有外显子,所有PCR产物分别进行单链构象多态性(SSCP)分析,通过患者与正常人带型的差异确定突变发生的外显子,对有差异SSCP带型的PCR产物进行直接DNA测序,找到突变位点。结果该家系先天性无虹膜表型明显与PAX6基因连锁;SSCP分析PAX6基因第9外显子PCR产物,显示患者均有异常带型出现,而家系正常人均无此异常带;测序结果显示突变位点为PAX6基因第9外显子c1080核苷酸C突变为T,使编码精氨酸的密码子突变为终止密码子。结论PAX6基因突变可导致先天性无虹膜。     

中国一先天性无虹膜家系PAX6基因新致病突变位点的筛查及验证

背景 先天性无虹膜是临床上罕见的先天性遗传性眼病.研究显示,配对盒转录因子6基因(PAX6)与先天性无虹膜症密切相关,但不同患者中PAX6基因的突变位点不同. 目的 对中国一常染色体显性遗传先天性无虹膜家系进行PAX6基因突变位点分析. 方法 于2014年8月在郑州大学第一附属医院收集一汉族先天性无虹膜家系,采集该家系9名成员及同期100名健康体检者的外周静脉血10 ml,采用标准酚-氯仿提取法提取基因组DNA,对PCR扩增产物进行测序、对比及突变分析.采用实时荧光定量PCR法验证和比较该家系中患病者与该家系表型正常者和健康对照者淋巴细胞中PAX6 mRNA的相对含量. 结果 该家系共3代9名成员,遗传方式符合常染色体显性遗传.家系中共5例患病者,成年患病者均表现为虹膜缺失和白内障,儿童患病者表现为无虹膜;其他4名家系成员表型正常.测序结果显示,家系患病者均存在11号染色体PAX6基因10号外显子的移码突变,第796位核糖核苷酸G缺失(c.796 del G),产生提前终止密码子,而家系正常成员及100名对照者均无此突变.实时荧光定量PCR结果显示,家系中患病者淋巴细胞中PAX6 mRNA表达水平比家系中正常成员约低50％,差异有统计学意义(Z=-2.449,P=0.016). 结论 PAX6基因c.796 del G为此先天性无虹膜家系的致病突变位点,扩增了PAX6基因突变谱.     

A Novel PAX6 Mutation in Chinese Patients with Severe Congenital Aniridia

Purpose: We identified a novel mutation in Paired Box gene 6 (PAX6) and characterized its associated clinical features of severe ocular malformation in a Chinese family with congenital aniridia. Methods: We studied two patients with aniridia from a Chinese family. All patients and noncarriers in this family underwent full ophthalmologic, general and urinary examinations. Total genomic DNA was isolated from peripheral blood of two aniridia patients. PAX6 levels were determined by PCR and its mutational status was determined by sequencing. Direct sequencing detected variations in PAX6. Results: Patients had bilateral congenital nystagmus, anterior polar cataract, absence of iris tissue, and foveal hypoplasia with severely reduced visual acuity. A novel heterozygous PAX6 mutation in exon 6 c.662G>A (p.W100X) was identified which created a premature termination codon. This observed sequence alteration was not found in 100 normal controls and has not been previously reported. Conclusions: We identified a novel PAX6 mutation in a family with severe ocular malformation. Our study expands the mutational spectrum of PAX6 and enriches our knowledge of the relationship between genotype and phenotype due to these mutations.     

PAX6 Analysis of Two Unrelated Families from the Arabian Peninsula with Classic Hereditary Aniridia

Introduction: Reports from around the world confirm that heterozygous PAX6 mutation is the major cause of hereditary aniridia (with a classic phenotype of iris hypoplasia, keratopathy, lens opacity, and foveal hypoplasia). However, genotype/phenotype reports are lacking from the Arabian Peninsula, a historically isolated region with a relatively high incidence of recessive disease and thus a potential for phenocopy and pseudodominance. The purpose of this study to assess for PAX6 mutation in two unrelated families with classic hereditary aniridia from the Arabian Peninsula. Methods: Interventional cases series of two unrelated affected Saudi Arabian families. Available family members underwent ophthalmic examination and venous blood sampling for PAX6 sequencing. Results: The pedigrees of both families suggested dominant (or pseudodominant) inheritance of the classic aniridia phenotype. Affected individuals in Family #1 were heterozygous for a novel frameshift PAX6 mutation (c.delA1294). Affected individuals in Family #2 had heterozygosity for a commonly-reported PAX6 nonsense mutation (p.Arg240X). Conclusions: PAX6 haploinsufficiency, the major cause of classic hereditary aniridia worldwide, is also associated with the phenotype in two different families from the Arabian Peninsula. Homozygosity by descent is not expected to affect genotype/phenotype correlation for the classic phenotype.     

A novel PAX6 gene mutation (P118R) in a family with congenital nystagmus associated with a variant form of aniridia

Background: A variety of PAX6 gene mutations were identified in patients with aniridia and/or allied ocular dysgenesis such as keratopathy, Peters’ anomaly, foveal hypoplasia, and nystagmus. To scrutinize the etiology of a four-generation Japanese family with autosomal dominant nystagmus associated with anterior and posterior segment anomalies, the PAX6 gene was examined. Patients and methods: A Japanese family showed a variant aniridia phenotype in four successive generations. Affected individuals had congenital nystagmus, microcornea with shortened axial length, superficial peripheral corneal opacification with pannus formation, dislocated pupil, and foveal hypoplasia. Analysis of the PAX6 gene mutation was performed in affected and unaffected individuals. Results: A novel missense mutation in the PAX6 gene was found in all affected individuals examined, but neither in unaffected individuals nor in unrelated healthy individuals. This mutation predicted a proline to arginine change at codon 118 (P118R) in the paired domain of PAX6 protein. Conclusion: The reported family illustrates that mutations in the PAX6 gene, in particular missense mutations, may manifest atypical clinical expression or forme fruste of aniridia. Received: 24 June 1999 Revised: 22 November 1999 Accepted: 23 November 1999     

中国一先天性无虹膜家系的遗传分析及PAX6基因突变位点的检测

背景 先天性无虹膜是双眼发病的遗传性疾病,目前的研究表明先天性无虹膜患者配对盒转录因子6(PAX6)基因突变位点具有多样性. 目的 通过目标序列捕获测序结合一代测序验证技术对1个中国先天性无虹膜家系进行基因突变位点的筛查和遗传分析.方法 采用横断面研究方法,本研究组于2016年3月纳入在郑州大学第一附属医院确诊的1个中国汉族先天性无虹膜家系,并对该家系成员进行致病突变基因检测.该家系全体成员均接受神经系统、口服葡萄糖耐量试验等全身体格检查以及眼科相关检查.采集现存家系所有成员前臂静脉血10 ml以提取基因组DNA,以先证者基因组DNA为模板行前房角发育异常致病基因的目标基因定点捕获测序分析,经与各基因库比对筛选出候选致病基因位点,采用PCR法对该家系成员行致病基因位点DNA片段扩增,采用Sanger测序技术在该家系除先证者以外的2例患病者和表型正常成员中进行候选致病基因验证. 结果 该家系共3代9名成员,Ⅰ1去世,现存8位成员,包括患病者3例(Ⅱ2及其子代Ⅲ1、Ⅲ2)和表型正常者5人,符合常染色体显性遗传模式.所有家系成员未发现神经系统异常,口服葡萄糖耐量试验结果均呈阴性.3例患病者视力均明显下降且不能矫正,眼压平均值为21 mmHg(1 mmHg=0.133 kPa),患者均存在虹膜完全缺如、角膜基质层混浊、眼球水平震颤、黄斑中心凹发育不良症状.此外,Ⅱ2患者存在左眼上睑下垂、右眼先天性白内障表现,Ⅲ2同时存在双眼先天性白内障、双侧晶状体不全脱位.先证者目标序列捕获测序分析及数据库比对显示,所有患病者PAX6基因第6号外显子上碱基替换c.183C＞A,经Sanger测序验证后证实突变基因与表型共分离. 结论 PAX6基因c.183C＞A突变是该先天性无虹膜家系的致病突变位点.     

A Novel CYP1B1 Mutation with Congenital Glaucoma and Total Aniridia

Purpose: Primary congenital glaucoma is a common disorder in the Middle East mainly caused by mutations in the the CYP1Bl gene. We report a family with three siblings that presented with recalcitrant childhood glaucoma, aniridia in two siblings with a novel CYP1B1 gene mutation.

Materials and methods: Review of pedigree, clinical history and clinical course of the family. Genetic testing in the affected family members.

Results: Three sisters presented with clinical findings of severe congenital glaucoma and a positive family history. Clinical examination of two of sisters revealed corneal scarring, bilateral aniridia with severe glaucoma that required multiple surgical procedures to control intraocular pressure. The third sibling presented with garden-variety primary congenital glaucoma. Genetic analysis revealed a novel CYP1B1 gene mutation (g.8291 C?>?T; p.S485F).

Conclusion: CYP1B1 mutation related congenital glaucoma can present with an extreme form of anterior segment dysgenesis that includes recalcitrant glaucoma, corneal opacification and aniridia.     

A Review of the Clinical and Genetic Aspects of Aniridia

Abstract

Aniridia classically presents with a bilateral congenital absence or malformation of the irides, foveal hypoplasia, and nystagmus, and patients tend to develop visually significant pre-senile cataracts and keratopathy. Additionally, they are at high risk for developing glaucoma. Classic aniridia can be genetically defined as the presence of a PAX6 gene deletion or loss-of-function mutation that results in haploinsufficiency. Variants of aniridia, which include a condition previously referred to as autosomal dominant keratitis, are likely due to PAX6 mutations that lead to partial loss of PAX6 function. Aniridia-associated keratopathy (AAK) is a progressive and potentially debilitating problem affecting aniridic patients. The current treatments for AAK are to replace the limbal stem cells through keratolimbal allograft (KLAL) with or without subsequent keratoplasty for visual rehabilitation, or to implant a Boston type 1 keratoprosthesis. Future therapies for AAK may be aimed at the genetic modification of corneal limbal stem cells.     

土家族中一个先天性无虹膜家系的临床特点和PAX6基因突变位点分析

目的：确认土家族中一个先天性无虹膜家系的PAX6基因致病突变并分析其临床特点。方法：实验研究。详细询问家族病史并对该家系中所有7 例成员（4 例患者,3 例正常人）进行详细的眼部检查,采集家系成员及100例（50例土家族人和50例汉族人）正常对照者的外周静脉血,提取DNA;对先证者PAX6基因的全部外显子进行PCR扩增及测序;对家系中所有成员和正常对照者进行PAX6基因突变位点的验证检测。结果：该家系中患者主要以虹膜缺损、白内障、眼球震颤、黄斑中心凹发育不良和角膜病变为主要临床表现,虹膜缺损轻重不一,角膜病变和白内障情况随年龄增加而加重。该家系的4 例患者均在第3 外显子与内含子3 交界处出现一个杂合突变（c.357+1G > A）,正常家系成员及正常对照者均无此突变。结论：该先天性无虹膜家系患者虹膜缺损程度不一。PAX6是该家系的致病基因,该家系患者PAX6基因的突变位点是杂合突变(c.357+1G > A）。     

先天性无虹膜症一家系PAX6基因Q310X突变的致病研究

目的 探讨先天性无虹膜症一家系的致病基因突变情况与发病机制.方法 采用病例对照研究方法.对该家系所有成员21人进行全面的眼科检查,同时进行家系调查并采集外周血样本,分离单个核细胞;用基因组DNA纯化试剂盒提取基因组DNA,以先证者DNA为模板聚合酶链反应法扩增转录因子PAX6基因全部14个外显子,用双脱氧末端终止法进行测序分析.结果 测序结果发现先证者Ⅲ2的PAX6基因在第11外显子上有Q310X(c.1378C>T)无义突变.它导致了第301位氨基酸密码子由CAA改变为TAX(Q301X),编码的谷氨酰胺突变为强终止密码子.对该家系中所有21名成员PAX6基因测序,结果发现所有10例患者都携带这一突变,而11名正常人则未检测到这一改变.结论 PAX6基因Q310X的无义突变所致PAX6蛋白翻译提前终止是此先天性无虹膜症家系的致病原因.     

Non-invasive anterior segment and posterior segment optical coherence tomography and phenotypic characterization of aniridia

Objective

To determine the value of optical coherence tomography (OCT) as a diagnostic tool in the critical evaluation of phenotypic variability seen in an aniridia family with a novel PAX6 mutation.

Design

Genetic and observational family study.

Participants

Three-generation family segregating autosomal dominant aniridia.

Methods

Ophthalmic examination included best-corrected visual acuity, slit-lamp biomicroscopy, direct and indirect ophthalmoscopy, tonometry, and OCT. PAX6 gene mutation analysis was carried out by direct sequencing of gene-specific PCR products and protein analysis by Western blot.

Results

Intrafamilial variable expressivity was seen between 4 affected family members. Phenotype differences between twin children suggested that this was due to modifier gene effects rather than environment. Anterior segment OCT demonstrated a range of iridocorneal angle abnormalities and corneal thickening in only 3, but ciliary body hypoplasia in all 4 affected patients. Posterior segment OCT demonstrated dome-shaped, hypoplastic macular profiles in the 2 affected children. Novel outer retinal changes were also seen, suggestive of a phototoxic retinopathy not previously recognized in aniridia. Ocular disease segregated with a novel PAX6 Q178X nonsense mutation with Western blot analysis suggesting that this led to haploinsufficiency of PAX6 protein.

Conclusions

Non-contact OCT imaging allowed for a more detailed assessment of anterior and posterior segment disease in children and adults with aniridia plus nystagmus. This led to the identification of novel features and highlights a practical, non-contact strategy well suited to genotype/phenotype studies and the longitudinal management of aniridic glaucoma in children.     

A novel PAX6 frameshift mutation in a kindred from Atlantic Canada with familial aniridia

BACKGROUND: Many mutations in PAX6, a member of a family of genes essential for normal development, have been described. We carried out a study to identify the mutation in PAX6 responsible for aniridia, an autosomal dominant disorder, in a kindred from Atlantic Canada. METHODS: Polymerase chain reaction amplification of coding exons, single-strand conformation polymorphism analysis and DNA sequencing. RESULTS: A novel deletion of an adenosine residue at position 1030 (1030delA) was detected. INTERPRETATION: The mutation responsible for aniridia in this kindred is expected to cause a frameshift in the PAX6 coding sequence and truncation of the homeodomain, which is essential for the function of the pax6 protein.     

三个先天性无虹膜症家系中PAX6基因的突变分析

背景 人类配对盒基因(PAX6)编码一个转录调节子,对眼和大脑形态的形成起关键作用.PAX6突变可导致许多先天性眼部发育异常,如先天性无虹膜症,通常为常染色体显性遗传方式. 目的 对三个先天性无虹膜症家系成员进行PAX6基因分析,探索这些家系发病的遗传基础. 方法 收集三个先天性无虹膜症家系的5例患病者和正常成员13名的外周血标本提取DNA,根据PAX6基因的序列设计4～ 13外显子序列,通过聚合酶链反应(PCR)扩增引物并测序,将目标序列与已发表的PAX6基因序列进行对比分析.结果 三个家系中共有5例患病者,在家系A中2例患者发现一个杂合突变(c.718 C＞T),导致第240位氨基酸由精氨酸突变为终止密码子(p.A rg240X),而其他正常表型者未发现此突变.家系B中的患病者和正常成员均未检测到PAX6基因的突变.家系C中1例患病者发现c.331 delG(p.Val111SerfsX13)的缺失突变,此单个碱基的缺失造成了移码突变,使PAX6蛋白羧基端的299个氨基酸缺失,而此家系的其他正常表型成员未发现此突变. 结论 家系A和家系C先天性无虹膜症的致病与PAX6基因的突变有关.     

PAX6基因突变至先天性无虹膜一家系的临床相关性研究

目的探讨PAX6基因突变引起先天性无虹膜症眼部及全身疾病发病的规律。方法对家系成员进行详细的视力检查、裂隙灯检查、前房角检查、眼底检查及眼压测量;应用核磁共振(MRI)技术对该家系的18例带有PAX6突变基因的患者和家系中6名正常人及人群中与该家系年龄段匹配的正常人进行脑部结构的扫描和应用CT技术进行脑结构的扫描;口服葡萄糖耐受实验;家系所有成员及与家系中患者年龄相当的对照组人员,空腹12 h以上,抽取静脉血6 mL,口服葡萄糖75 g后分别抽取0.5 h和2 h的静脉血各6 mL,分离血清,对所有血样测定葡萄糖水平。家系成员进行鼻内窥镜检查和CT扫描。结果该家系患者除无虹膜外,还合并多种眼部疾病,随年龄增长眼部并发症逐渐增多,视力逐渐下降甚至失明;家系大部分患者表现有不同程度的脑部结构异常,且年龄越小的患者其胼胝体的变性萎缩越轻,随年龄增长胼胝体的变性萎缩加重;该家系中所有30岁以上的患者均有不同程度的糖耐量异常甚至糖尿病;该家系患者有鼻结构异常或鼻窦炎,患病率明显高于正常人群。结论PAX6基因突变所致先天性无虹膜症不是独立的眼科疾病,而是以无虹膜为首发症状,同时合并多种全身疾病的一种综合征。     

Molecular characterization of Axenfeld-Rieger spectrum and other anterior segment dysgeneses in a sample of Mexican patients

ABSTRACT

Background: Anterior segment dysgenesis (ASD) and Axenfeld-Rieger spectrum (ARS) are mainly due to PITX2 and FOXC1 defects, but it is difficult in some patients to differentiate among PITX2-, FOXC1-, PAX6- and CYP1B1-related disorders. Here, we set out to characterize the pathogenic variants (PV) in PITX2, FOXC1, CYP1B1 and PAX6 in nine unrelated Mexican ARS/ASD patients and in their available affected/unaffected relatives.

Materials and methods: Automated Sanger sequencing of PITX2, FOXC1, PAX6 and CYP1B1 was performed; those patients without a PV were subsequently analyzed by Multiplex Ligation-dependent Probe Amplification (MLPA) for PITX2, FOXC1 and PAX6. Missense variants were evaluated with the MutPred, Provean, PMUT, SIFT, PolyPhen-2, CUPSAT and HOPE programs.

Results: We identified three novel PV in PITX2 (NM_153427.2:c.217G>A, c.233T>C and c.279del) and two in FOXC1 [NM_001453.2:c.274C>T (novel) and c.454T>A] in five ARS patients. The previously reported FOXC1 c.367C>T or p.(Gln123*) variant was identified in a patient with ASD. The ocular phenotype related to FOXC1 included aniridia, corneal opacity and early onset glaucoma, while an asymmetric ocular phenotype and aniridia were associated with PITX2. No gene rearrangements were documented by MLPA analysis, nor were any PV identified in PAX6 or CYP1B1.

Conclusions: Heterozygous PV in the PITX2 and FOXC1 genes accounted for 66% (6/9) of the ARS/ASD cases. The absence of PAX6 or CYP1B1 abnormalities could reflect our small sample size, although their analysis could be justified in ARS/ASD patients that present with congenital glaucoma or aniridia.