Similar miRNomic signatures characterize the follicular fluids collected after follicular and luteal phase stimulations in the same ovarian cycle
Danilo Cimadomo, Ramona Carmelo, Elvira Immacolata Parrotta, Stefania Scalise, Gianluca Santamaria, Erminia Alviggi, Maria Teresa De Angelis, Gianmarco Sarro, Alberto Vaiarelli, Roberta Venturella, Laura Rienzi, Fulvio Zullo, Filippo Maria Ubaldi, Giovanni Cuda
Journal of Assisted Reproduction and Genetics – 2019 Nov 7 – Received: 12 July 2019 / Accepted: 4 October 2019 – https://doi.org/10.1007/s10815-019-01607-6
Purpose To detect putative differences in the miRNomic profile of follicular fluids collected after follicular-phase-stimulation (FPS-FFs) and paired luteal-phase-stimulation (LPS-FFs) in the same ovarian cycles (DuoStim).
Methods Exploratory study at a private IVF center and University involving FPS-FFs and paired-LPS-FFs collected from 15 reduced ovarian reserve and advanced maternal age women undergoing DuoStim (n = 30 paired samples).
The samples were combined in 6 paired pools (5 samples each) and balanced according to maternal age and number of cumulus-oocyte-complexes. Micro-RNAs were isolated and sequenced.
Four miRNAs were then selected for further validation on 6 single pairs of FPS-FFs and LPS-FFs by qPCR.
Results Forty-three miRNAs were detected in both FPS-FFs and paired-LPS-FFs after sequencing and no statistically significant differences were reported.
Thirty-three KEGG pathways were identified as regulated from the detected miRNAs. Four miRNAs (miR-146b, miR-191, miR-320a, and miR-483) were selected for qPCR validation since consistently expressed in our samples and possibly involved in the regulation/establishment of a healthy follicular environment.
Again, no significant differences were reported between FPS-FFs and paired-LPS-FFs, also when the analysis was corrected for maternal age and number of cumulus- oocyte-complexes in generalized linear models.
Conclusions These data complement the embryological, chromosomal, and clinical evidence of equivalence between FPS and LPS published to date.
Keywords: Follicular fluid . DuoStim . Double stimulation . Luteal phase stimulation . miRNA