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Abstract：
OBJECTIVE: To investigate the mutations in patients with Axenfeld-Rieger syndrome (ARS) and the pattern of PITX2-related tooth agenesis.
METHODS: Whole-exome sequencing (WES) and copy number variation (CNV) array were used to screen the mutations in four ARS probands. After Sanger sequencing and quantitative polymerase chain reaction (qPCR) validation, secondary structure prediction and dual-luciferase assay were employed to investigate the functional impact. Eighteen PITX2-mutated patients with definite dental records were retrieved from our database and literatures, and the pattern of PITX2-related tooth agenesis was analyzed.
RESULTS: A novel de novo segmental deletion of chromosome 4q25 (GRCh37/hg19 chr4:111, 320, 052-111, 754, 236) encompassing PITX2 and three novel PITX2 mutations c.148C > T, c.257G > A, and c.630insCG were identified. Preliminary functional studies indicated the transactivation capacity of mutant PITX2 on Distal-less homeobox 2 (DLX2) promoter was compromised. The maxillary teeth showed significantly higher rate of agenesis (57.94%) than the mandibular teeth (44.05%). The most often missing teeth were upper lateral incisors (83.33%) and upper second premolars (69.44%). Teeth with the least agenesis rate were the lower second molars (19.44%) and lower first molars (8.33%).
CONCLUSIONS: We identified a novel 4q25 microdeletion including PITX2 and three novel PITX2 mutations, and statistically analyzed the PITX2-related tooth agenesis pattern.
METHODS: Whole-exome sequencing (WES) and copy number variation (CNV) array were used to screen the mutations in four ARS probands. After Sanger sequencing and quantitative polymerase chain reaction (qPCR) validation, secondary structure prediction and dual-luciferase assay were employed to investigate the functional impact. Eighteen PITX2-mutated patients with definite dental records were retrieved from our database and literatures, and the pattern of PITX2-related tooth agenesis was analyzed.
RESULTS: A novel de novo segmental deletion of chromosome 4q25 (GRCh37/hg19 chr4:111, 320, 052-111, 754, 236) encompassing PITX2 and three novel PITX2 mutations c.148C > T, c.257G > A, and c.630insCG were identified. Preliminary functional studies indicated the transactivation capacity of mutant PITX2 on Distal-less homeobox 2 (DLX2) promoter was compromised. The maxillary teeth showed significantly higher rate of agenesis (57.94%) than the mandibular teeth (44.05%). The most often missing teeth were upper lateral incisors (83.33%) and upper second premolars (69.44%). Teeth with the least agenesis rate were the lower second molars (19.44%) and lower first molars (8.33%).
CONCLUSIONS: We identified a novel 4q25 microdeletion including PITX2 and three novel PITX2 mutations, and statistically analyzed the PITX2-related tooth agenesis pattern.
摘要：
目的：研究Axenfeld-Rieger综合征（ARS）患者的突变和PITX2相关牙齿发育不全的模式。
方法：使用全外显子组测序（WES）和拷贝数变异（CNV）阵列来筛选四个ARS先证者的突变。经过Sanger测序和定量聚合酶链反应（qPCR）验证，采用二级结构预测和双荧光素酶测定来研究功能影响。从我们的数据库和文献中检索到18例具有明确牙科记录的PITX2突变患者，并分析了PITX2相关牙齿发育不全的规律。
结果：染色体4q25（GRCh37/hg19chr4：111，320，052-111，754，236）的新的从头分段缺失，包括PITX2和三个新的PITX2突变c.148C>T，c.257G>A，和c.630insCG被确认。初步功能研究表明，突变体PITX2在无远端同源异型框2（DLX2）启动子上的反式激活能力受损。上颌牙齿的发育不全率（57.94％）明显高于下颌牙齿（44.05％）。最常见的牙齿缺失是上侧切牙（83.33％）和上第二前磨牙（69.44％）。发育率最低的牙齿是下第二磨牙（19.44％）和下第一磨牙（8.33％）。
结论：我们鉴定了一个新的4q25微缺失，包括PITX2和三个新的PITX2突变，并对PITX2相关牙齿发育不全模式进行统计分析。
方法：使用全外显子组测序（WES）和拷贝数变异（CNV）阵列来筛选四个ARS先证者的突变。经过Sanger测序和定量聚合酶链反应（qPCR）验证，采用二级结构预测和双荧光素酶测定来研究功能影响。从我们的数据库和文献中检索到18例具有明确牙科记录的PITX2突变患者，并分析了PITX2相关牙齿发育不全的规律。
结果：染色体4q25（GRCh37/hg19chr4：111，320，052-111，754，236）的新的从头分段缺失，包括PITX2和三个新的PITX2突变c.148C>T，c.257G>A，和c.630insCG被确认。初步功能研究表明，突变体PITX2在无远端同源异型框2（DLX2）启动子上的反式激活能力受损。上颌牙齿的发育不全率（57.94％）明显高于下颌牙齿（44.05％）。最常见的牙齿缺失是上侧切牙（83.33％）和上第二前磨牙（69.44％）。发育率最低的牙齿是下第二磨牙（19.44％）和下第一磨牙（8.33％）。
结论：我们鉴定了一个新的4q25微缺失，包括PITX2和三个新的PITX2突变，并对PITX2相关牙齿发育不全模式进行统计分析。
