Molecular pathology of congenital insensitivity to pain with anhidrosis due to genetic defects of the receptor tyrosine kinase for nerve growth factor

神经生长因子受体酪氨酸激酶遗传缺陷导致先天性疼痛不敏感伴无汗症的分子病理学

• 批准号: 13672378
• 负责人: INDO Yasuhiro
• 金额: $ 2.3万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672378/
• 关键词: congenital insensitivity to pain with anhidrosis; hereditary sensory and autonomic neuropathy type IV; nerve growth factor; nerve growth factor receptor; receptor tyrosine kinase; TRKA; NTRK1; 遺伝性感覚自立神経性ニューロパチーIV型; 片親性ダイソミー

1. We have analyzed the TRKA gene derived from 23 Japanese and 8 foreign patients with congenital insensitivity to pain with anhidrosis (CIPA) and detected responsible mutations in all these patients. We also report the characterization of intragenic polymorphic sites and describe the haplotypic associations of alleles at these sites in Japanese CIPA families. More than 50% of CIPA chromosomes share the frameshift mutation (R548fs) that we described earlier. This mutation apparently shows linkage disequilibrium with a rare haplotype in normal chromosomes, strongly suggesting that it is a common founder mutation.2. Uniparental disomy (UPD) is defined as the presence of a chromosome pair that derives from only one patient in a diploid individual. We have observed a male CIPA patient with non-Mendelian inheritance. He had a homozygous mutation at the TRKA locus on chromosome 1. Haplotype analysis of the TRKA locus and allelotype analyses of whole chromosome 1 revealed that the chromosome … More pair was exclusively derived from his father. Non-maternity was excluded by analyses of autosomes other than chromosome 1. Thus, we have identified a complete paternal isodisomy for chromosome 1 as the cause of reduction to homozygosity of the TRKA gene mutation, leading to CIPA.3. We have demonstrated that an intronic branch-site (IVS7-33T>A) mutation in the TRKA gene causes aberrant splicing in vitro by the exon-trap analysis. We also reported 11 putative missense mutations in 32 CIPA families from various ethnic groups. Here we have introduced the corresponding mutations into the TRKA cDNA and examined NGF-stimulated autophosphorylation. Two mutants (L93P and L213P) in the extracellular domain were aberrantly processed and showed diminished autophosphorylation in neuronal cells. Five mutants (G516R, G571R, R643W, R648C and G708S) in the tyrosine kinase domain were processed as wild-type TRKA but showed significantly diminished autophosphorylation in both neuronal and non-neuronal cells. In contrast, R85S and H598Y; G607V detected previously as double and triple mutations, are probably polymorphisms in a particular ethnic background. The other putative mutant D668Y might be a rare polymorphism or might impair the function of TRKA without compromising autophosphorylation. Mutated residues in the tyrosine kinase domain are conserved in various receptor tyrosine kinases (RTKs) and probably contribute to critical function of these proteins. Thus, naturally occurring TRKA missense mutations with loss-of-function provide considerable insight into the structure-function relationship in the RTK family. Less

1. 我们分析了23名日本和8名外国先天性无汗性疼痛不敏感（CIPA）患者的TRKA基因，并在所有这些患者中检测到相关突变。我们还报道了日本CIPA家族基因内多态性位点的特征，并描述了这些位点上等位基因的单倍型关联。超过50%的CIPA染色体共享我们之前描述的移码突变（R548fs）。该突变明显表现出与正常染色体中一种罕见的单倍型的连锁不平衡，强烈表明它是一种常见的奠基者突变。单倍体二体（UPD）被定义为在二倍体个体中仅来自一个患者的染色体对的存在。我们观察了一位非孟德尔遗传的男性CIPA患者。他在1号染色体上的TRKA位点有一个纯合突变。TRKA位点的单倍型分析和1号染色体的等位型分析表明，1对染色体完全来自父亲。除1号染色体外的常染色体分析排除非母性。因此，我们已经确定了1号染色体的完全父系同工二体是TRKA基因突变纯合性降低的原因，导致cipa。我们已经通过外显子陷阱分析证明了TRKA基因的内含子分支位点（IVS7-33T>A）突变导致体外异常剪接。我们还报道了来自不同种族的32个CIPA家族的11个假定的错义突变。在这里，我们将相应的突变引入TRKA cDNA，并检测了ngf刺激的自磷酸化。细胞外域的两个突变体（L93P和L213P）在神经元细胞中被异常加工并表现出自磷酸化减少。酪氨酸激酶结构域的5个突变体（G516R、G571R、R643W、R648C和G708S）被处理为野生型TRKA，但在神经元和非神经元细胞中均表现出显著的自磷酸化降低。相反，R85S和H598Y；以前检测到的G607V为双突变和三突变，可能是特定种族背景下的多态性。另一个推测的突变D668Y可能是一种罕见的多态性，或者可能在不影响自磷酸化的情况下损害TRKA的功能。酪氨酸激酶结构域的突变残基在各种受体酪氨酸激酶（RTKs）中是保守的，并且可能对这些蛋白的关键功能起作用。因此，具有功能丧失的自然发生的TRKA错义突变为RTK家族的结构-功能关系提供了相当大的见解。少

项目成果

犬童康弘: "先天性無痛無汗症 小児科(第2版) (白木、前川 監修) (伊藤、大関、岡田、近藤、杉本、田澤、田村、埜中、原田、福嶋 編)"医学書院. 1534-1535 (2002)

犬户泰宏：《先天性无痛无汗症儿科学》（第 2 版）（白木和前川编）（伊藤、大关、冈田、近藤、杉本、田泽、田村、野中、原田和福岛编辑）《伊学书院》1534-1535 年。 (2002)

犬童康弘: "先天性無痛無汗症"生体の科学. 50. 379-380 (1999)

Yasuhiro Inudo：“先天性无痛无汗症”生物科学 50. 379-380 (1999)。

Y.Miura: "Complete paternal uniparental isodisomy for chromosome 1 revealed by mutation analyses of the TRKA (NTRK1) gene encoding a receptor tyrosine kinase for nerve growth factor in a patient with congenital insensitivity to pain with anhidrosis"Human

Y.Miura：“对先天性疼痛不敏感伴无汗症患者的神经生长因子受体酪氨酸激酶 TRKA (NTRK1) 基因进行突变分析，揭示了 1 号染色体的完全父本单亲异构体”人类

犬童康弘: "先天性無痛無汗症の分子病態から見た交感神経と感覚神経の分化・生存とアポトーシス"自律神経. 39. 53-60 (2002)

Yasuhiro Inudo：“从先天性无痛无汗症的分子病理学角度观察交感神经和感觉神经的分化、存活和凋亡”自主神经学 39. 53-60 (2002)。

E.Toscano: "Multisystem involvement in congenital insensitivity to pain with anhidrosis (CIPA), a nerve growth factor receptor (Trk A)-related disorder"Neuropediatrics. 31. 39-41 (2000)

E.Toscano：“多系统参与先天性疼痛不敏感伴无汗症 (CIPA)，一种神经生长因子受体 (Trk A) 相关疾病”神经儿科。