Gene Expression Profiles of Human Granulosa Cells Treated With Bioequivalent Doses of Corifollitrophin Alfa or Recombinant Human Follicle-Stimulating Hormone

Using recombinant DNA technologies, a new chimeric gene containing the coding sequences of the follicle stimulating hormone (FSH) β-subunit and the C-terminal peptide of the human chorionic gonadotrophin (hCG) β-subunit have been designed to generate a new gonadotrophin named corifollitrophin alfa. Corifollitrophin alfa has longer elimination half-life and a slower rate of absorption compared with FSH, which makes corifollitrophin alfa a long-acting hormone employed as a substitute of the recombinant FSH (recFSH) in the controlled ovarian stimulation (COS). The purpose of this study is to compare the gene expression profiles elicited by bioequivalent doses of corifollitrophin alfa or recFSH in primary cultures of human granulosa cells (hGCs). Both gonadotrophins exert their functions by binding FSH receptors (FSHRs), activating signaling pathways that increase the cyclic adenosine monophosphate (cAMP) intracellular content. Bioequivalence has been defined as the dose/duration of gonadotrophin treatment able to promote the same amount of intracellular cAMP. Primary cultured hGCs were treated with different doses of either gonadotrophin and the cAMP was measured after different incubation times to establish the bioequivalence. Results obtained by comparing the bioequivalent treatments, showed that corifollitrophin alfa is more effective than recFSH in inducing aromatase gene expression after 6 and 24 hours from the initial stimulation in agreement with its long-acting characteristic’s. Primary cultures of human granulosa cells from five different patients were treated with bioequivalent doses of corifollitrophin alfa or rhFSH then washed to remove the unbuond hormone. Controls and gonadotrophin-stimulated samples were processed for total RNA extraction and hybridization on Affymetrix microarrays after 24 hours of incubation.