The Transcriptome Landscape change of Human Granulosa Cells under gonadotropins Revealed by Small bulk Cell RNA Sequencing. 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). Overall design: One leading pre-ovulatory follicle per subject was used from three natural-cycle and four Gns-stimulated patients.