O-26: Effects of Recombinant-LH Supplementation on The Proteomic Profile of Follicular Fluid from Poor Responder Patients: Focus on Follicular Growth Factors and Oocyte Maturity Markers
Poor and fragmentary data was available regarding the effects of recombinant-LH (r-LH) in in-vitro granulosa and theca cells of human origin. No data was available from in vivo studies regarding the effects of r-LH supplementation on SCF,EGF,ERK-1/2 and AKT-1 pathways in the follicular fluid of older-poor-responder women undergoing IVF cycle. Evidence from in-vitro and animal studies seems to confirm that r-LH, in addition increasing cAMP levels (involved in steroidogenesis), activates the ERK-1/2 (proliferation) and AKT-1 (anti-apoptotic) pathways. The aim of the study project is to understand if the r-LH supplementation during IVF (in-vitro-fertilization) cycles in poor responder patients may influence the pathways involved in follicular growth and oocyte maturity. To understand which follicular signaling pathways are influenced by r-LH supplementation and finally to understand if these factors may explain the clinical advantages of r-LH supplementation in older poor-responder-patients.
Materials and methods
We recruited 28 poor responder patients older than 42 years. All patients underwent COH (controlled-ovarian-hyperstimulation) using r-FSH 300 IU/ day (recombinant-FSH) alone in the first cycle (s-COH group) and using r-FSH (300 IU/day) with r-LH (150 IU/day) supplementation (ex-COH group) in the second cycle. We compared follicular concentrations of SCF, EGF, Erk 1-2, p-Erk 1-2, Akt-1 and p-Akt-1 between the two groups. Follicular levels of SCF, EGF, Erk 1-2, p-Erk-1/2, Akt-1, p-Akt-1 were detected using appropriate ELISA-Kit and reported in pg/mL, ng/ mL or Unit/mL, according to the manufacturer’s indications.
Follicular levels (56 samples) of EGF, Erk 1-2, p- Erk-1/2, Akt-1, p-Akt-1 were significantly different between s-COH versus ex-COH, with the exception of SCF. In detail, mean value of EGF was 9.40 ± 2.92 vs. 11.75 ± 3.95 pg/mL (P<0.05), ERK 1-2 184.82 ± 50.15 vs. 332.14 ± 111.35 pg/ mL (P<0.001), p-ERK-1/2 20.89 ± 3.41 vs. 40.18 ± 10.37 U/ mL (P<0.001), AKT-1 5.35 ± 2.45 vs. 10.42 ± 2.08 ng/mL (P<0.001), p-AKT-1 28.07 ± 8.98 vs. 42.36 ± 10.06 U/mL (P<0.001), SCF 830.25 ± 364.09 vs. 735.43 ± 300.39 pg/mL (p: n. s.). The increasing intra-follicular levels of proteins (particularly in their active conformation) involved in cellular proliferation and anti-apoptotic pathways may explain the better clinical outcome observed after r-LH supplementation. The absence of significant variations in SCF levels confirmed both that this pathway is activated only by FSH stimulation and that differences collected in other pathways is generated by r-LH signaling.
In poor-responder patients, r-LH supplementation during IVF influences the pathways involved in follicular growth and oocyte maturity. The treatment significantly increases follicular levels of EGF, ERK-1/2 and AKT-1 and particularly the availability of the phosphorylated forms (active forms). These evidences explain the improvements in qualitative and quantitative ovarian response. This is the first in vivo study reporting these evidences, partially demonstrated by invitro/ experimental studies on animals.