Abstract

Central precocious puberty (CPP) is a pediatric endocrine disorder characterized by the premature activation of hypothalamic gonadotropin-releasing hormone (GnRH) neurons. At present, CPP researches predominantly rely on animal models, which cannot fully recapitulate the endocrine properties of human GnRH neurons. Herein, we aimed to generate GnRH neurons from induced pluripotent stem cells (iPSCs) derived from the peripheral blood of CPP patient, thereby establishing a novel human cell-based model of CPP. Using a classical three-step in vitro differentiation protocol, iPSCs from both CPP patient and healthy adult females (serving as the healthy control (HC) group) were differentiated into GnRH neurons. Quantitative real-time polymerase chain reaction, immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) were employed to assess the expression of GNRH1 and microtubule-associated protein 2 (MAP2), as well as the secretion levels of GnRH and γ-aminobutyric acid (GABA). At day 27, 28 and 29, GnRH neurons derived from the CPP group exhibited significantly elevated GNRH1 mRNA levels, along with higher fluorescence intensities of GNRH1 and MAP2 compared to those from the HC group. ELISA further demonstrated that GnRH concentrations in CPP were significantly higher at all examined time points compared with the HC group. In contrast, GABA concentrations were reduced in CPP. These findings indicated that CPP patient-derived GnRH neurons displayed intrinsically accelerated maturation and GnRH hypersecretion, accompanied by attenuated GABAergic inhibitory function. Together, this study has successfully established a CPP cell model based on iPSCs derived from CPP patient, providing a novel experimental platform for in-depth exploration of the pathogenesis and drug intervention for CPP in the future.