Secretory Profile Analysis of Human Granulosa Cell Line Following Gonadotropin Stimulation

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

doi:10.3390/ijms26094108

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Open AccessArticle

Secretory Profile Analysis of Human Granulosa Cell Line Following Gonadotropin Stimulation

by

Francesca Mancini

^1,†

,

Emanuela Teveroni

^1,†

,

Michela Cicchinelli

²

,

Federica Iavarone

^2,3

,

Anna Laura Astorri

¹,

Giuseppe Maulucci

^4,5

,

Cassandra Serantoni

^4,5

,

Duaa Hatem

^4,5

,

Daniela Gallo

^6,‡,

Carla Di Nardo

⁷,

Andrea Urbani

^2,3,

Alfredo Pontecorvi

⁷,

Domenico Milardi

^1,7,*

and

Fiorella Di Nicuolo

¹

International Scientific Institute Paul VI, Università Cattolica del Sacro Cuore, Fondazione Policlinico Universitario A. Gemelli IRCCS, 00168 Rome, Italy

²

Department of Basic Biotechnological Sciences, Intensivological and Perioperative Clinics, Università Cattolica del Sacro Cuore, 00168 Rome, Italy

³

Clinical Chemistry, Biochemistry and Molecular Biology Operations (UOC), Fondazione Policlinico Universitario A. Gemelli IRCCS, 00168 Rome, Italy

⁴

Department of Neuroscience, Section of Biophysics, Università Cattolica del Sacro Cuore, 00168 Rome, Italy

⁵

Fondazione Policlinico Universitario A. Gemelli IRCCS, 00168 Rome, Italy

⁶

Dipartimento Scienze della Salute della Donna, del Bambino e di Sanità Pubblica, Fondazione Policlinico Universitario A. Gemelli IRCCS, Lgo A. Gemelli 8, 00168 Roma, Italy

⁷

Division of Endocrinology, Fondazione Policlinico Universitario A. Gemelli IRCCS, 00168 Rome, Italy

^*

Author to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

^‡

Current address: Department of Toxicology & Pathology Sciences, European Research Biology Center (ERBC), Via Tito Speri 12/14, 00071 Pomezia, Italy.

Int. J. Mol. Sci. 2025, 26(9), 4108; https://doi.org/10.3390/ijms26094108

Submission received: 27 February 2025 / Revised: 16 April 2025 / Accepted: 17 April 2025 / Published: 25 April 2025

(This article belongs to the Special Issue Central and Local Modulators of Reproduction and Fertility: An Update (4th Edition))

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Abstract

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.

Keywords: KGN; granulosa cells; secretomic analysis; FSH; hCG; semaphorin

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MDPI and ACS Style

Mancini, F.; Teveroni, E.; Cicchinelli, M.; Iavarone, F.; Astorri, A.L.; Maulucci, G.; Serantoni, C.; Hatem, D.; Gallo, D.; Di Nardo, C.; et al. Secretory Profile Analysis of Human Granulosa Cell Line Following Gonadotropin Stimulation. Int. J. Mol. Sci. 2025, 26, 4108. https://doi.org/10.3390/ijms26094108

AMA Style

Mancini F, Teveroni E, Cicchinelli M, Iavarone F, Astorri AL, Maulucci G, Serantoni C, Hatem D, Gallo D, Di Nardo C, et al. Secretory Profile Analysis of Human Granulosa Cell Line Following Gonadotropin Stimulation. International Journal of Molecular Sciences. 2025; 26(9):4108. https://doi.org/10.3390/ijms26094108

Chicago/Turabian Style

Mancini, Francesca, Emanuela Teveroni, Michela Cicchinelli, Federica Iavarone, Anna Laura Astorri, Giuseppe Maulucci, Cassandra Serantoni, Duaa Hatem, Daniela Gallo, Carla Di Nardo, and et al. 2025. "Secretory Profile Analysis of Human Granulosa Cell Line Following Gonadotropin Stimulation" International Journal of Molecular Sciences 26, no. 9: 4108. https://doi.org/10.3390/ijms26094108

APA Style

Mancini, F., Teveroni, E., Cicchinelli, M., Iavarone, F., Astorri, A. L., Maulucci, G., Serantoni, C., Hatem, D., Gallo, D., Di Nardo, C., Urbani, A., Pontecorvi, A., Milardi, D., & Di Nicuolo, F. (2025). Secretory Profile Analysis of Human Granulosa Cell Line Following Gonadotropin Stimulation. International Journal of Molecular Sciences, 26(9), 4108. https://doi.org/10.3390/ijms26094108

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Secretory Profile Analysis of Human Granulosa Cell Line Following Gonadotropin Stimulation

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