A novel de novo TP63 mutation in whole-exome sequencing of a Syrian family with Oral cleft and ectrodactyly.
A novel de novo TP63 mutation in whole-exome sequencing of a Syrian family with Oral cleft and ectrodactyly.
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DOI：
10.1002/mgg3.2179
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发表时间：
2023-08
影响因子：
2
通讯作者：
中科院分区：
文献类型：
--
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口裂和缺指是常见的异质性出生缺陷。我们在一个叙利亚家庭中进行了全外显子组测序（WES）分析。先证者同时出现口面裂和缺指畸形，但未出现缺指畸形、外胚层发育不良和唇腭裂综合征中常见的外胚层发育不良-3。一位只有唇裂的叔叔去世了，无法进行分析。变异注释，孟德尔的不一致性，并在已知的裂缝基因的新变种进行了检查。候选变体使用桑格测序进行验证，并通过敲除斑马鱼中的tp 63基因来评估其在斑马鱼发育期间的作用来评估致病性。确定了28个候选的新发事件，其中一个是已知的唇裂和缺趾基因TP 63（c.956G > T，p.Arg319Leu），并通过桑格测序证实。 TP 63突变与多发性常染色体显性口面裂和肢体畸形疾病相关在该患者中观察到的p.Arg319Leu突变是新发的，但也是新的。同一密码子中的两个已知突变（c.956G > A，p.（Arg 319 His; rs 121908839，c.955C > T），p.Arg319Cys）引起缺指畸形，提供了突变该密码子是有害的证据。虽然这种TP 63突变是患者临床表现的最佳候选者，但它是否负责整个表型尚不清楚。tp 63敲除斑马鱼的产生和表征显示在受精后3天（dpf）头部坏死和破裂。胚胎表型不能通过注射斑马鱼或人信使RNA（mRNA）来拯救。需要进一步的功能分析，以确定表型的比例是由于这种突变。一种新的从头突变TP 63基因被发现在患者口面裂和缺指。我们发现，在斑马鱼中突变该基因会导致头部破裂的严重表型，并且我们无法在注射斑马鱼或人mRNA后挽救该表型。目前还不清楚这种突变是导致整个表型还是仅仅导致缺指畸形，需要进一步的研究。
Oral clefts and ectrodactyly are common, heterogeneous birth defects. We performed whole‐exome sequencing (WES) analysis in a Syrian family. The proband presented with both orofacial clefting and ectrodactyly but not ectodermal dysplasia as typically seen in ectrodactyly, ectodermal dysplasia, and cleft lip/palate syndrome‐3. A paternal uncle with only an oral cleft was deceased and unavailable for analysis. Variant annotation, Mendelian inconsistencies, and novel variants in known cleft genes were examined. Candidate variants were validated using Sanger sequencing, and pathogenicity assessed by knocking out the tp63 gene in zebrafish to evaluate its role during zebrafish development. Twenty‐eight candidate de novo events were identified, one of which is in a known oral cleft and ectrodactyly gene, TP63 (c.956G > T, p.Arg319Leu), and confirmed by Sanger sequencing. TP63 mutations are associated with multiple autosomal dominant orofacial clefting and limb malformation disorders. The p.Arg319Leu mutation seen in this patient is de novo but also novel. Two known mutations in the same codon (c.956G > A, p.(Arg319His; rs121908839, c.955C > T), p.Arg319Cys) cause ectrodactyly, providing evidence that mutating this codon is deleterious. While this TP63 mutation is the best candidate for the patient's clinical presentation, whether it is responsible for the entire phenotype is unclear. Generation and characterization of tp63 knockout zebrafish showed necrosis and rupture of the head at 3 days post‐fertilization (dpf). The embryonic phenotype could not be rescued by injection of zebrafish or human messenger RNA (mRNA). Further functional analysis is needed to determine what proportion of the phenotype is due to this mutation. A novel de novo mutation in the TP63 gene was found in a patient with an orofacial cleft and ectrodactyly. We showed that mutating this gene in zebrafish caused a severe phenotype where the head ruptured and we were unable to rescue the phenotype upon injection of either zebrafish or human mRNA. It is unknown whether the mutation is responsible for the entire phenotype or just the ectrodactyly and further research is warranted.
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