Molecular analysis of a mutated FSH receptor detected in a patient with spontaneous ovarian hyperstimulation syndrome
Authors: Sayaka Uchida, Hiroshi Uchida, Tetsuo Maruyama, Takashi Kajitani, Hideyuki Oda, Kaoru Miyazaki, Maki Kagami, Yasunori Yoshimura
Journal: PLoS One (2013)
MeSH terms: Mutation, Missense, Neoplasm Proteins / genetics, Ovarian Hyperstimulation Syndrome / genetics, Pituitary Neoplasms / genetics, Receptors, FSH / genetics, Case Reports, Clinical Trial, Research Support, Non-U.S. Gov't, Adult, Amino Acid Substitution, Cell Line, Female, Follicle Stimulating Hormone / blood, Follicle Stimulating Hormone / genetics, Follicle Stimulating Hormone / therapeutic use, Hormone Replacement Therapy / methods, Humans, MAP Kinase Signaling System / drug effects, MAP Kinase Signaling System / genetics, Middle Aged, Neoplasm Proteins / blood, Ovarian Hyperstimulation Syndrome / blood, Ovarian Hyperstimulation Syndrome / drug therapy, Phosphatidylinositol 3-Kinases / genetics, Phosphatidylinositol 3-Kinases / metabolism, Pituitary Neoplasms / blood, Pituitary Neoplasms / drug therapy, Protein-Tyrosine Kinases / genetics, Protein-Tyrosine Kinases / metabolism, Receptors, FSH / blood, Neoplasm Proteins, Receptors, FSH, Follicle Stimulating Hormone, Phosphatidylinositol 3-Kinases, Protein-Tyrosine Kinases

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.