The elucidation of pathoenic mechanism of cranio-maxillo-facial anomdy and the applicaton for gene diagnosis

本研究的目的是系统地分析一系列先天性形态异常的候选基因。颅缝闭锁是一种由一条或多条颅缝过早融合引起的病理状态。最近，成纤维细胞生长因子受体（FGFR）基因的突变已在综合征性颅缝闭闭中被检测到。成纤维细胞生长因子（FGF）和FGFR调节多种细胞活性、细胞生长和胚胎发育。首先，我们采用聚合酶链反应（PCR）和直接测序的方法检测了日本颅缝闭锁患者（Crouzon综合征9例，Apert综合征6例，无综合征患者颅骨畸形3例）FGFR2的核苷酸序列。结果显示，在Crouzon综合征患者中，在第7外显子的密码子252、290和第9外显子的密码子342、354处存在FGFR2杂合突变，在Apert综合征患者中，分别在5例和1例患者中检测到Ser252Trp和Pro253Arg。在1例Crouzon、所有舟头畸形和健康个体中检测到更多的突变。迄今为止，在许多高加索患者中报道了综合征性颅缝闭闭中FGFR2的序列分析，而在日本仅研究了几个病例。目前对IS患者的研究证实，无论种族和环境如何，日本患者发生的一系列突变与高加索患者相似。日本Crouzon患者的突变频率为82%（9/11例）。日本Apert患者中S252W: P253R的比值为5:1。此外，在日本的Apert患者中，Ser252Trp突变的腭裂并发症发生率为60%，Pro253Arg突变的腭裂并发症发生率为0/2，并指分分别为4.90分和5.50分。同源盒型（HOX）基因在胚胎期的形态发生中起着重要的作用。其次，我们怀疑Apert综合征的手部异常可能是HOXA13和HOXD13基因的原因，因此我们对它们进行了分析。此外，我们还分析了MSX1（HOX7）基因与Apert综合征腭裂的关系。我们检测到Apert组与健康对照组之间存在多种多态性和频率不同的可能性。下一个主题，锁骨颅骨发育不良（CCD）是一种常染色体显性的人类骨骼疾病，其特征是锁骨发育不全或再发育不全、颅骨缝合线宽、牙齿多余、身材矮小和其他骨骼疾病。近年来，在CCD患者中检测到核心结合因子（CBFAI）基因的多种突变。CBFAI基因是转录因子家族中的一员。我们报告了一例日本的CCD患者，伴有开放性缝合线、锁骨发育不全和短指畸形，并伴有寰枢脱位。我们对CBFAI基因进行了序列分析，并在第3外显子检测到R225W的错义突变。少

The purpose of this study is to systematically analysis for candidate gene of a series of congenital morphological anomaly.Craniosynostosis is the pathologic condition that results from premature fusion of one or more cranial sutures. Recently, mutations of the fibroblast growth factor receptor (FGFR) genes have been detected in syndromic craniosynostosis. The fibroblast growth factor(FGF) and FGFR regulate multiple cellular activities, cell growth and embryonal development. Firstly, we examined nucleotide sequences of FGFR2 in Japanese craniosynostosis patients (Crouzon syndrome : 9 cases, Apert syndrome : 6 cases and scaphocephaly : 3 cases as non-syndromic patients) by polymerase chain reaction (PCR) followed by direct sequencing methods. The results demonstrated FGFR2 heterozygous mutations at codons 252, 290 of exon 7, and at codon 342, 354 of exon 9 in Crouzon syndromes, in Apert syndrome patients, Ser252Trp and Pro253Arg were detected in five and one patients, respectively. No m … More utation was detected in one case of Crouzon, all cases of scaphocephaly and healthy individuals. Thus far sequence analysis of FGFR2 in syndromic craniosynostosis has been reported in many Caucasian patients, whereas in Japanese only several cases have been studied. The present study with IS patients confirmed mat a similar series of mutations occur in Japanese patients as in Caucasian patients regardless of ethnicity and environment. The frequency of the mutation was 82% (9/11 cases) in Japanese Crouzon patients. The ratio of S252W : P253R was 5 : 1 in Japanese Apert patients. Morever, in Japanese Apert patients, complication rate of cleft palate was 60% for mutation of Ser252Trp and 0/2 of patients for Pro253Arg, with their syndactyly score being 4.90 and 5.50, respectively.Homeobox (HOX) gene has the role which is important for the morphogenesis in the embryonic stage. Secondary, hand anomaly of Apert syndrome was suspected as cause of HOXA13 and HOXD13 gene, so we analyzed them. Moreover, we also analyzed MSX1(HOX7) gene as cause of cleft palate in Apert syndrome. We detected several polymorphism and mere were possibility of different frequency between Apert group and healthy control group.Next subject, cleidocranial dysplasia (CCD) is an autosomal dominant human bone disease characterized by hypoplastic or aplastic clavicles, wide cranial sutures, supernumerary teeth, short stature, and other skeletal disorders. Recently, various mutations of the core binding factor (CBFAI) gene have been detected in CCD patients. The CBFAI gene is a member of the runt family of transcription factors. We experienced one Japanese case of CCD with open sutures, hypoplasia of clavicles and brachydactyly, combined with atlant-axis dislocation. We performed the sequence analysis of the CBFAI gene and detected a missense mutation of R225W in exon 3. Less