Mancini Francesca, Teveroni Emanuela, Cicchinelli Michela, Iavarone Federica, Astorri Anna Laura, Maulucci Giuseppe, Serantoni Cassandra, Hatem Duaa, Gallo Daniela, Di Nardo Carla, Urbani Andrea, Pontecorvi Alfredo, Milardi Domenico, Di Nicuolo Fiorella
International Scientific Institute Paul VI, Università Cattolica del Sacro Cuore, Fondazione Policlinico Universitario A. Gemelli IRCCS, 00168 Rome, Italy.
Department of Basic Biotechnological Sciences, Intensivological and Perioperative Clinics, Università Cattolica del Sacro Cuore, 00168 Rome, Italy.
Int J Mol Sci. 2025 Apr 25;26(9):4108. doi: 10.3390/ijms26094108.
Granulosa cell (GC) differentiation, stimulated by FSH and LH, drives oocyte maturation and follicle development. FSH promotes GC proliferation, and LH triggers ovulation. In clinical practice, hCG is used to mimic LH. Despite various controlled ovarian stimulation (COS) protocols employing exogenous gonadotropins and GnRH analogs to prevent premature ovulation, their effectiveness and safety remain debated. To identify markers predicting a positive treatment response, the secretome of gonadotropin-stimulated GC using the human granulosa-like tumor cell line (KGN) via proteomics was analyzed. Additionally, a novel 2D-FFT quantitative method was employed to assess cytoskeleton fiber aggregation and polymerization, which are critical processes for GC differentiation. Furthermore, the activation of key kinases, focal adhesion kinase (FAK), and Rho-associated coiled-coil-containing protein kinase 1 (ROCK-1), which are implicated in cytoskeleton dynamics and hormone signaling, was evaluated. The proteomic analysis revealed significant modulation of proteins involved in extracellular matrix organization, steroidogenesis, and cytoskeleton remodeling. Notably, the combined FSH/hCG treatment led to a dynamic upregulation of the semaphorin pathway, specifically semaphorin 7A. Finally, a significant reorganization of the cytoskeleton network and signaling was detected. These findings enhance our understanding of folliculogenesis and suggest potential novel molecular markers for predicting patient responses to gonadotropin stimulation.
在促卵泡生成素（FSH）和促黄体生成素（LH）的刺激下，颗粒细胞（GC）分化驱动卵母细胞成熟和卵泡发育。FSH促进GC增殖，而LH触发排卵。在临床实践中，人绒毛膜促性腺激素（hCG）被用于模拟LH。尽管有各种使用外源性促性腺激素和促性腺激素释放激素（GnRH）类似物来防止过早排卵的控制性卵巢刺激（COS）方案，但其有效性和安全性仍存在争议。为了确定预测阳性治疗反应的标志物，通过蛋白质组学分析了使用人颗粒样肿瘤细胞系（KGN）的促性腺激素刺激的GC的分泌蛋白质组。此外，采用了一种新颖的二维快速傅里叶变换（2D - FFT）定量方法来评估细胞骨架纤维的聚集和聚合，这是GC分化的关键过程。此外，还评估了参与细胞骨架动力学和激素信号传导的关键激酶，粘着斑激酶（FAK）和含Rho相关卷曲螺旋的蛋白激酶1（ROCK - 1）的激活情况。蛋白质组学分析揭示了参与细胞外基质组织、类固醇生成和细胞骨架重塑的蛋白质的显著调节。值得注意的是，联合FSH/hCG治疗导致信号素通路，特别是信号素7A的动态上调。最后，检测到细胞骨架网络和信号传导的显著重组。这些发现加深了我们对卵泡发生的理解，并提示了预测患者对促性腺激素刺激反应的潜在新分子标志物。
