Craniofacial microsomia (CFM), also known as the oculo-auriculo-vertebral spectrum, is a congenital disorder characterized by hypoplasia of the mandible and external ear due to tissue malformations originating from the first and second branchial arches. However, distinguishing it from other syndromes of branchial arch abnormalities is difficult, and causal variants remain unidentified in many cases. In this report, we performed an exome sequencing analysis of a Brazilian family with CFM. The proband was a 12-month-old boy with clinical findings consistent with the diagnostic criteria for CFM, including unilateral mandibular hypoplasia, microtia, and external auditory canal abnormalities. A heterozygous de novo nonsense variant (c.713C>G, p.S238*) in PUF60 was identified, which was predicted to be pathogenic in silico. PUF60 has been reported as a causal gene in Verheij syndrome, but not in CFM. Although the boy showed craniofacial abnormalities and developmental delay that overlapped with Verheij syndrome, the facial asymmetry with unilateral hypoplasia of the mandible observed in this case did not match the previously reported phenotypes of PUF60 variants. Our findings expand the phenotypic range of PUF60 variants that cover CFM and Verheij syndrome.

Heterozygous variants in the Poly(U) Binding Splicing Factor 60kDa gene (PUF60) have been associated with Verheij syndrome, which has the key features of coloboma, short stature, skeletal abnormalities, developmental delay, palatal abnormalities, and congenital heart and kidney defects. Here, we report five novel patients from unrelated families with PUF60-related disorders exhibiting novel genetic and clinical findings with three truncating variants, one splice-site variant with likely reduced protein expression, and one missense variant. Protein modeling of the patient\'s missense variant in the PUF60 AlphaFold structure revealed a loss of polar bonds to the surrounding residues. Neurodevelopmental disorders were present in all patients, with variability in speech, motor, cognitive, social-emotional and behavioral features. Novel phenotypic expansions included movement disorders as well as immunological findings with recurrent respiratory, urinary and ear infections, atopic diseases, and skin abnormalities. We discuss the role of PUF60 in immunity with and without infection based on recent organismic and cellular studies. As our five patients showed less-severe phenotypes than classical Verheij syndrome, particularly with the absence of key features such as coloboma or palatal abnormalities, we propose a reclassification as PUF60-related neurodevelopmental disorders with multi-system involvement. These findings will aid in the genetic counseling of patients and families.

Verheij syndrome (VRJS) is a craniofacial spliceosomopathy with a wide phenotypic spectrum. Haploinsufficiency of the poly-uridine binding splicing factor 60 gene (PUF60) and its loss-of-function (LOF) variants are involved in VRJS. We evaluated a human fetus with congenital heart defects and preaxial polydactyly. Clinical data were obtained from the medical record. Whole-exome sequencing (WES) was used to explore the potential genetic etiology, and the detected variant verified using Sanger sequencing. Functional studies were performed to validate the pathogenic effects of the variant. Using trio-WES, we identified a novel PUF60 variant (NM_078480.2; c.1678 T > A, p.*560Argext*204) in the pedigree. Bioinformatic analyses revealed that the variant is potentially pathogenic, and functional studies indicated that it leads to degradation of the elongated protein and subsequently PUF60 LOF, producing some VRJS phenotypes. These findings confirmed the pathogenicity of the variant. This study implicates PUF60 LOF in the etiopathogenesis of VRJS. It not only expands the PUF60 variant spectrum, and also provides a basis for genetic counseling and the diagnosis of VRJS. Although trio-WES is a well-established approach for identifying the genetic etiology of rare multisystemic conditions, functional studies could aid in verifying the pathogenicity of novel variants.

PUF60-related developmental disorder (also referred to as Verheij syndrome), resulting from haploinsufficiency of PUF60, is associated with multiple congenital anomalies affecting a wide range of body systems. These anomalies include ophthalmic coloboma, and congenital anomalies of the heart, kidney, and musculoskeletal system. Behavioral and intellectual difficulties are also observed. While less common than other features associated with PUF60-related developmental disorder, for instance hearing impairment and short stature, identification of specific anomalies such as ophthalmic coloboma can aid with diagnostic identification given the limited spectrum of genes linked with this feature. We describe 10 patients with PUF60 gene variants, bringing the total number reported in the literature, to varying levels of details, to 56 patients. Patients were recruited both via locally based exome sequencing from international sites and from the DDD study in the United Kingdom. Eight of the variants reported were novel PUF60 variants. The addition of a further patient with a reported c449-457del variant to the existing literature highlights this as a recurrent variant. One variant was inherited from an affected parent. This is the first example in the literature of an inherited variant resulting in PUF60-related developmental disorder. Two patients (20%) were reported to have a renal anomaly consistent with 22% of cases in previously reported literature. Two patients received specialist endocrine treatment. More commonly observed were clinical features such as: cardiac anomalies (40%), ocular abnormalities (70%), intellectual disability (60%), and skeletal abnormalities (80%). Facial features did not demonstrate a recognizable gestalt. Of note, but remaining of unclear causality, we describe a single pediatric patient with pineoblastoma. We recommend that stature and pubertal progress should be monitored in PUF60-related developmental disorder with a low threshold for endocrine investigations as hormone therapy may be indicated. Our study reports an inherited case with PUF60-related developmental disorder which has important genetic counseling implications for families.

Verheij syndrome (VRJS) is a rare craniofacial spliceosomopathy presenting with craniofacial dysmorphism, multiple congenital anomalies and variable neurodevelopmental delay. It is caused by single nucleotide variants (SNVs) in PUF60 or interstitial deletions of the 8q24.3 region. PUF60 encodes a splicing factor which forms part of the spliceosome. To date, 36 patients with a sole diagnosis of VRJS due to disease-causing PUF60 SNVs have been reported in peer-reviewed publications. Although the depth of their phenotyping has varied greatly, they exhibit marked phenotypic heterogeneity. We report 10 additional unrelated patients, including the first described patients of Khmer, Indian, and Vietnamese ethnicities, and the eldest patient to date, with 10 heterozygous PUF60 variants identified through exome sequencing, 8 previously unreported. All patients underwent deep phenotyping identifying variable dysmorphism, growth delay, neurodevelopmental delay, and multiple congenital anomalies, including several unique features. The eldest patient is the only reported individual with a germline variant and neither neurodevelopmental delay nor intellectual disability. In combining these detailed phenotypic data with that of previously reported patients (n = 46), we further refine the known frequencies of features associated with VRJS. These include neurodevelopmental delay/intellectual disability (98%), axial skeletal anomalies (74%), appendicular skeletal anomalies (73%), oral anomalies (68%), short stature (66%), cardiac anomalies (63%), brain malformations (48%), hearing loss (46%), microcephaly (41%), colobomata (38%), and other ocular anomalies (65%). This case series, incorporating three patients from previously unreported ethnic backgrounds, further delineates the broad pleiotropy and mutational spectrum of PUF60 pathogenic variants.

Verheij syndrome (VRJS) is a rare microdeletion syndrome of chromosome 8q24.3 that is characterized by severe growth retardation, microcephaly, vertebral anomalies, joint laxity/dislocation, psychomotor retardation, cardiac and renal defects, and dysmorphic facial features. Pathogenic variants of PUF60 (Poly-U Binding Splicing Factor 60 kDa) have been found to cause VRJS. Here we present a Turkish patient with Verheij syndrome who has typical facial dysmorphic features and renal and cardiac abnormalities, scoliosis, tethered cord, and mild intellectual disability.
This is a case report of a 11-year-old female child who presented with Verheij syndrome. Blood samples were collected from the patient and the family. We performed whole exome sequencing was used to identify potential genetic mutations. We also used 3-dimensional protein structure analysis to identify the effect of the mutation.
A de-novo in-frame variant (c.449_457delCAAAGGGGG; p.Ala150_Phe152del) of the PUF60 gene was identified by whole exome sequencing. According to ACMG guidelines in 2015, the mutation is classified as pathogenic and it has been reported in the clinvar database. Results of in-silico prediction software tools predicted the mutation was pathogenic. Protein structure analysis showed that the three residues affected by the in-frame deletion form could lead to impaired stability and function of the PUF60 protein.
To date, 25 patients have been reported with PUF60 mutations in the medical literature. In this article, we report a patient with VRJS who had the unusual findings of tethered cord syndrome and renal abnormalities. As far as we know, this is the first patient from Turkey who has been diagnosed with Verheij syndrome.

暂无摘要。

BACKGROUND: Verheij syndrome is a rare microdeletion syndrome of chromosome 8q24.3 that harbors PUF60, SCRIB, and NRBP2 genes. Subsequently, loss of function mutations in PUF60 have been found in children with clinical features significantly overlapping with Verheij.
METHODS: Here we present the first Chinese Han patient with a de novo nonsense variant (c.1357C > T, p.Gln453*) in PUF60 by clinical whole exome sequencing. The 5-year-old boy presents with dysmorphic facial features, intellectual disability, and growth retardation but without apparent cardiac, renal, ocular, and spinal anomalies.
CONCLUSIONS: Our finding contributes to the understanding of the genotype and phenotype in PUF60 related disorder.

Objective: To investigate the clinical and genetic characteristics of a Chinese boy with Verheij syndrome and review the literature. Methods: The clinical and genetic data of a Chinese boy with Verheij syndrome, who was admitted to the Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology in May 2017 were analyzed. Original papers on Verheij syndrome published up to January 2018 were retrieved at PubMed, Human Gene Mutation Database (HGMD), Online Mendelian Inheritance in Man(OMIM), CNKI and WanFang databases by using the key words \"Verheij syndrome\" and \"PUF60\" . Results: The male patient (at the age of 14 years and 3 months) visited us because of growth retardation for 13 years. Atrial septal defect was repaired at the age 3. Congenital amblyopia and hyperopia were diagnosed at the age 4. On physical examination, serious growth retardation and delayed psychomotor development was noted. His height was 142.5 cm (-3.26 SDS). He had poor academic performance at school. Facial features included: webbed neck, hypertelorism, down-slanting palpebral fissures, long philtrum, thin upper lip, and high palate. Palmar crease was found in the right hand. His bone age was 10 years. Growth hormone stimulation test indicated partial growth hormone deficiency (growth hormone (GH) peak 6.63 μg/L). The level of insulin like growth factor 1 (IGF1) and insulin like growth factor binding protein 3 (IGFBP3) was lower than normal, 73.20 μg/L and 2 500 μg/L respectively. Abdominal ultrasound showed that the volumes of bilateral kidneys were small. The size of the left and right kidney was 8.5 cm × 3.3 cm and 8.4 cm × 4.3 cm respectively. Karyotype was normal (46, XY). MRI of pituitary showed partial empty sella turcica. Ten genes associated with Noonan syndrome (PTPN11, SOS1, RASA2, KRAS, RAF1, NRAS, SHOC2, BRAF, RIT1, A2ML1) were analyzed and no genetic mutations were found. Whole exome-sequencing analysis identified a de novo heterozygous frame shift mutation of PUF60 gene (c.931_934del, P.P.T 311Qfs*47). According to ACMG guidelines in 2015, the mutation is pathogenic and has not been reported in the above databases. Conclusions: This is the first case report of Verheij syndrome caused by mutation of PUF60 gene in Chinese population. It is difficult to discriminate Verheij syndrome from Noonan syndrome, both have clinical manifestations such as severe growth retardation, psychomotor retardation, and congenital heart disease. In addition to Noonan syndrome, PUF60 genetic analysis was recommended for avoiding missed diagnosis with such clinical manifestations of patients.
目的： 总结Verheij综合征的临床及遗传学分析特点。 方法： 对2017年5月华中科技大学同济医学院附属同济医院儿科收治的1例Verheij综合征患儿的临床资料及基因检测结果进行总结分析，并以\"Verheij syndrome\"\"PUF60\"和\"Verheij综合征\"\"PUF60基因\"为检索词，分别检索2018年1月前的PubMed、人类基因组突变数据库（HGMD）、在线人类孟德尔遗传数据库（OMIM）、中国知网数据库（CNKI）和万方数据库，对Verheij综合征病例进行文献复习。 结果： 患儿男，14岁3月龄，因\"生长落后13年\"就诊。出生体重2.9 kg，3岁时在外院行\"房间隔缺损修补术\"，4岁时发现\"先天性弱视及远视\"。身高142.5 cm，标准差积分－3.26。精神运动发育迟缓，学习成绩不佳。颈蹼，眼距宽、双侧外眼角下斜，人中长，腭弓高，后发际低；右手通贯掌。骨龄落后于实际年龄；垂体内分泌功能检测结果提示部分性生长激素（GH）缺乏（GH峰值6.63 μg/L），血胰岛素样生长因子1（IGF1）（73.20 μg/L）及胰岛素样生长因子结合蛋白3（IGFBP3）（2 500 μg/L）低于正常；肾脏B超左肾8.5 cm×3.3 cm，右肾8.4 cm×4.3 cm；染色体核型46,XY；垂体磁共振成像提示垂体高度约4 mm，存在部分空蝶鞍。内分泌基因Panel（与Noonan综合征相关的10个基因）分析，均未发现基因变异或缺失。全外显子基因检测发现患儿存在PUF60基因杂合变异（c.931_934del，p. p.T311Qfs*47）。患儿父母该位点均无变异。经Sanger测序进行验证及父母来源分析，提示为新发突变。根据2015年美国医学遗传学与基因组学学会指南，该移码突变为致病突变，且在上述数据库中尚未见报道。 结论： 首次报道中国人群PUF60基因变异导致的Verheij综合征。患儿有严重的生长发育落后、精神运动发育迟缓、特殊面部特征、先天性心脏病、肾脏发育不良等，临床与Noonan综合征难以鉴别。对具有上述临床表现的患儿除注意Noonan综合征相关基因检测外，还应注意PUF60基因分析，以免漏诊。.