Study on the pathogenesis for syndrome of resistance to thyroid hormone using cultured human skin fibroblasts.

利用培养的人皮肤成纤维细胞研究甲状腺激素抵抗综合征的发病机制。

基本信息

批准号：
04671470
负责人：
MURATA Yoshiharu
金额：
$ 1.28万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04671470/
关键词：
Thyroid hourmone Thyroxine Hormone resistance Fibroblast

项目摘要

The syndrome of generalized resistance to thyroid hormone(GRTH)is characterized by reduced clinical and biochemical manifestations of thyroid hormone action relative to the circulating hormone levels. Several reports demonstrated abnormalities in beta isoform of T_3 receptor gene(c-erbA beta). However, a question how the mutation of c-erbA beta cause the clinical manifestation of the syndrome still remaied to be clarified. The current research aimed to uncover the pathogenesis of a syndrome of generalized resistance to thyroid hormone(GRTH)using cultured skin fibroblasts obtained from the patients. In the fitst year of investigation, we tried to clarify how the mutation in c-erbA beta affects the expression of other isoform of T_3 receptor gene(c-erbA alpha). We utilized reverse transcription-polymerase chain reaction(RT-PCR)to determine c-erbA mRNAs. We pick up two cases in family A which exibits one single nucleotide substitution in one allel of c-erbA beta gene. We also examined the exprtession of c-erbA a gene in fibroblasts from family B in which most of the coding region of c-erbA beta gene is deleted in both alleles. The result indicated that normal and mutated c-erbA beta genes are equally expressed in family A.In the case of family B, no c-erbA beta mRNA was detected. In both famity A and B, c-erbA alpha was expressed like in fibroblasts from normal subjects. From these results indicated hat the mutation does not affect the expression of normal c-erbA gene. Thus, we conculuded the pathogenesis of GRTH is unlikeky due to the abnormal expression of a normal allele of c-erbA beta and of c-erbA alpha. We next examined whether the type of mutation is related to the responsiveness to T_3 when the expressions of mRNAs for T_3 responsive genes are used as a marker of T_3 action. At first, we used the collagenase and the fibronetin as a marker, however, we gofound that these mRNA expression were altered by other factors than T<@2

对甲状腺激素（GRTH）的广义抗性综合征的特征是相对于循环激素水平，甲状腺激素作用的临床和生化表现降低。几份报告表明，T_3受体基因（C-ERBAβ）的β同工型异常。然而，一个问题C-ERBAβ的突变如何导致该综合征的临床表现仍被阐明。目前的研究旨在使用从患者获得的培养的皮肤成纤维细胞使用普遍性抗甲状腺激素（GRTH）的综合性发病机理。在调查的一年中，我们试图阐明C-Erba beta中的突变如何影响T_3受体基因（C-Erba alpha）其他同工型的表达。我们利用逆转录聚合酶链反应（RT-PCR）来确定C-ERBA mRNA。我们在家庭A中捡起两个病例，这些病例可以在C-Erbaβ基因的一个Allel中超出一个单核苷酸取代。我们还研究了来自家庭B的成纤维细胞中C-Erba A基因的拔出，在该基因中，C-Erba Beta基因的大多数编码区都在两个等位基因中都被删除。结果表明，正常和突变的c-Erbaβ基因在家族中平均表达。在家庭B的情况下，未检测到C-ErbaβmRNA。在Famity A和B中，C-Erba Alpha在正常受试者的成纤维细胞中表达。从这些结果表明HAT突变不会影响正常C-ERBA基因的表达。因此，由于c-erbaβ和c-erba alpha的正常等位基因的异常表达，我们结识了GRTH的发病机理不同。接下来，当将T_3响应基因的mRNA表达式用作T_3动作的标记时，我们接下来研究了突变类型是否与对T_3的响应能力有关。最初，我们使用胶原酶和纤烯素用作标记，但是，我们竭尽全力，这些mRNA表达被其他因素以外的其他因素改变了