The Transcriptome Landscape change of Human Granulosa Cells under gonadotropins Revealed by Small bulk Cell RNA Sequencing

STUDY QUESTION: Can superovulation dose of gonadotropins (Gns) affect the transcriptome of granulosa cells and follicular fluid (FF) hormone levels? SUMMARY ANSWER: Cell cycle- and meiosis-associated genes were down-regulated by Gns stimulation, whereas immune- and cytokine-associated genes were up-regulated. Hormone levels were also affected by Gns stimulation. WHAT IS KNOWN ALREADY: Superovulation treatment had adverse effects on maturation, spindle function, and chromosomal aneuploidy in oocytes. STUDY DESIGN, SIZE, DURATION: One leading pre-ovulatory follicle per subject was used from three natural-cycle and four Gns-stimulated patients. PARTICIPANTS/MATERIALS, SETTING, METHODS: Granulosa cells and FF were collected from the same leading follicle from each patient. RNA was extracted from granulosa cells and subjected to deep sequencing and analysis. Follicle-stimulating hormone (FSH), estradiol (E2), androstenedione (AND), testosterone (T), luteinizing hormone (LH), and progesterone (P) levels in FF were measured by immunoassays. Student s t test was used for statistical analysis. MAIN RESULTS AND ROLE OF CHANCE: A total of 715 genes were up-regulated, and 287 genes were down-regulated, in the Gns-stimulated group relative to the Control group. Gene Ontology analysis revealed that the down-regulated genes were enriched in cell cycle and meiosis pathways, primarily those associated with follicle or oocyte maturation and quality. On the other hand, the up-regulated genes were enriched in functions related to immunity and cytokine Ccytokine receptor interactions. In stimulated vs. natural-cycle follicles, E2 and LH levels were significantly reduced (P < 0.001). One leading pre-ovulatory follicle per subject was used from three natural-cycle and four Gns-stimulated patients.