Molecular analysis of a mutated FSH receptor detected in a patient with spontaneous ovarian hyperstimulation syndrome.
Molecular analysis of a mutated FSH receptor detected in a patient with spontaneous ovarian hyperstimulation syndrome.
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DOI：
10.1371/journal.pone.0075478
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Yoshimura Y
中科院分区：
文献类型：
--
作者：
Uchida S;Uchida H;Maruyama T;Kajitani T;Oda H;Miyazaki K;Kagami M;Yoshimura Y
自发性卵巢过度刺激综合征（sOHSS）是一种罕见的事件，可能是由FSH产生的垂体腺瘤（FSHoma）、FSH受体（FSHR）的激活突变以及妊娠或甲状腺功能减退症时FSHR对hCG和TSH升高的交叉反应性引起的。本研究的目的是调查是否存在异常FSHR的妇女与sOHSS和非手术诊断的FSHoma，其血清FSH水平和FSH生物活性接近正常。患者的FSHR基因测序显示了一个杂合的新的错义突变c。1536 G>A导致氨基酸取代M512 I。我们询问这种突变FSHR是否影响FSHR介导的信号通路，包括cAMP/蛋白激酶A（PKA）、磷脂酰肌醇-3激酶（PI 3 K）/蛋白激酶B（AKT）和v-src肉瘤（Schmidt-Ruppin A-2）病毒癌基因同源物激酶（SRC）/ p42/p44细胞外信号调节蛋白激酶（ERK 1/2）。因此，用FSH处理表达野生型（FSHRwt）、突变型FSHR（FSHRmt）或两者（FSHRwt/mt）的293 T细胞，并进行细胞内cAMP的测量、cAMP诱导的CRE（cAMP反应元件）介导的荧光素酶测定和磷酸化PI 3 K和ERK 1/2的免疫印迹分析。FSHRwt、FSHRmt和FSHwt/mt细胞间荧光素酶活性和ERK 1/2磷酸化水平无差异。然而，与FSHwt细胞相比，FSHRmt细胞在FSH刺激后cAMP产生和PI 3 K磷酸化水平显著降低，ERK 1/2磷酸化水平不变。此外，FSH处理不会引起FSHwt/mt细胞中的PI 3 K磷酸化。这些结果表明，本文鉴定的新型错义M512 I FSHR突变不参与FSHR介导的信号传导途径的超活化，而是参与FSH介导的PI 3 K/AKT途径的低活化。因此，这项研究表明，这种新的突变FSHR的一个新的功能特性，然而，这可能不参与sOHSS的发病机制，在这个FSHoma患者。
Spontaneous ovarian hyperstimulation syndrome (sOHSS) is a rare event that may result from a FSH-producing pituitary adenoma (FSHoma), activating mutations of the FSH receptor (FSHR), and cross-reactivity of the FSHR to elevated hCG and TSH in the setting of pregnancy or hypothyroidism. The objective of this study was to investigate whether an aberrant FSHR was present in a woman with sOHSS and a non-surgically diagnosed FSHoma whose serum FSH levels and FSH bioactivity were nearly normal. Sequencing of the patient’s FSHR gene revealed a heterozygous novel missense mutation c. 1536G>A resulting in an amino acid substitution M512I. We asked whether this mutant FSHR affected FSHR-mediated signaling pathways involving cAMP/protein kinase A (PKA), phosphatidylinositol-3 kinase (PI3K)/protein kinase B (AKT) and v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog kinase (SRC)/ p42/p44 extracellular signal-regulated protein kinases (ERK1/2). Thus, 293T cells expressing wild-type (FSHRwt), the mutant FSHR (FSHRmt), or both (FSHRwt/mt) were treated with FSH and subjected to measurements of intracellular cAMP, cAMP-induced CRE (cAMP response element)-mediated luciferase assays and immunoblot analyses of phosphorylated PI3K and ERK1/2. There were no differences in luciferase activities or phosphorylation levels of ERK1/2 among FSHRwt, FSHRmt cells and FSHwt/mt cells. However, FSHRmt cells showed a significant reduction in both cAMP production and PI3K phosphorylation levels with unchanged phosphorylation of ERK1/2 upon FSH stimulation in comparison to FSHwt cells. Also, FSH treatment did not provoke PI3K phosphorylation in FSHwt/mt cells. These results indicate that the novel missense M512I FSHR mutation identified herein did not participate in hyperactivation of FSHR-mediated signaling pathways but rather in hypoactivation of the FSH-mediated PI3K/AKT pathway. Thus, this study demonstrates a new functional property of this novel mutatnt FSHR, which, however, might not be involved in the pathogenesis of sOHSS in this FSHoma patient.
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