Effects and plasma proteomic analysis of GLP-1RA versus CPA/EE, in combination with metformin, on overweight PCOS women: a randomized controlled trial

Abstract

Purpose: Polycystic ovary syndrome (PCOS) is characterized by reproductive dysfunctions and metabolic disorders. This study aims to compare the therapeutic effectiveness of glucagon-like peptide-1 receptor agonist (GLP-1RA) + Metformin (Met) versus cyproterone acetate/ethinylestradiol (CPA/EE) + Met in overweight PCOS women and identify potential proteomic biomarkers of disease risk in women with PCOS.

Methods: In this prospective, open-label randomized controlled trial, we recruited 60 overweight PCOS women into two groups at a 1:1 ratio to receive CPA/EE (2 mg/day: 2 mg cyproterone acetate and 35-μg ethinylestradiol,) +Met (1500 mg/day) or GLP-1 RA (liraglutide, 1.2-1.8 mg/day) +Met (1500 mg/day) for 12 weeks. The clinical effectiveness and adverse effects were evaluated, followed by plasma proteomic analysis and verification of critical biomarkers by ELISA.

Results: Eighty(80%) patients completed the study. Both interventions improved menstrual cycle, polycystic ovaries, LH(luteinizing hormone) and HbA1c(hemoglobin A1c) levels after the 12-week treatment. GLP-1RA + Met was more effective than CPA/EE + Met in reducing body weight, BMI (Body Mass Index), and waist circumference, FBG(fasting blood glucose), AUCI(area under curve of insulin),TC (Total Cholesterol), IL-6(Interleukin-6) and improving insulin sensitivity, and ovulation in overweight women with PCOS, with acceptable short-term side effects. CPA/EE + Met was more effective in improving hyperandrogenemia, including T(total testosterone), LH, LH/FSH(Luteinizing hormone/follicle-stimulating hormone), SHBG(sex hormone-binding globulin) and FAI (free androgen index). By contract, GLP-1RA+Met group only improved LH. Plasma proteomic analysis revealed that the interventions altered proteins involved in reactive oxygen species detoxification (PRDX6, GSTO1, GSTP1, GSTM2), platelet degranulation (FN1), and the immune response (SERPINB9).

Conclusions: Both CPA/EE+Met and GLP-1RA + Met treatment improved reproductive functions in overweight PCOS women. GLP-1RA + Met was more effective than CPA/EE + Met in reducing body weight, BMI, and waist, and improving metabolism, and ovulation in overweight women with PCOS, with acceptable short-term side effects. CPA/EE + Met was more effective in reducing hyperandrogenemia. The novel plasma biomarkers PRDX6, FN1, and SERPINB9, might be indicators and targets for PCOS treatment. TRIAL REGISTRATION CLINICALTIALS.

Gov trial no: NCT03151005. Registered 12 May, 2017, https://clinicaltrials.gov/ct2/show/NCT03151005 .

Keywords: Glucagon-like Peptide-1 Receptor Agonist (GLP-1 RA); Metformin; PCOS (Polycystic Ovary Syndrome); Proteomics Analysis.

Fig. 1

Study flowchart. PCOS polycystic ovary syndrome, GLP-1RA glucagon-like peptide-1 receptor agonist, CPA cyproterone acetate, EE ethinylestradiol

Fig. 2

Proteomic analysis of plasma from PCOS pa tients treated with CPA/EE+Met or GLP-1RA+Met. Volcano plots of differentially expressed plasma proteins from patients prior to or after treatment with CPA/EE+Met (A) or GLP-1RA+Met (B). Red and blue dots represent significantly up- or downregulated protein expression, respectively (fold change ≥1.5 and P value < 0.05, moderated paired t test by limma, n = 3). Gray dots show nonsignificantly changed proteins. (C) Gene ontology (biological process) enrichment analysis of differentially expressed proteins from the two groups. The rich factor was calculated by the number of proteins enriched in a specific item divided by the total number of proteins in that term. (D) Network showing interactions between drugs (orange diamonds), differentially expressed proteins (green circles), and selected enriched functional terms (blue squares). (E) Abundance heatmap of sixteen selected proteins prior to and after treatment

Fig. 3

Changes in the characteristics of clinical parameters were observed after CPA/EE+Met and GLP-1RA + Met treatments. These changes included weight (A), Waist (B), T (C), LH (D), LH/FSH (E), SHBG (F), FAI (G), Regular menstruation (H), HAb1c (I), HOMA-IR (J), TC (K), IL-6 (L), FINS (M), n = 30. The data are presented as mean ± SEM. * P value < 0.05, ** P value < 0.01, *** P value < 0.001 (vs. before treatment in each group). Changes in FINS (M), after CPA/EE+Met and GLP-1 RA+Met treatment * P value < 0.05 (vs. before treatment in GLP-1RA + Met)

Fig. 4

ELISA validation of plasma candidate protein marker levels. Plasma levels of PRDX6 (A), GSTO1 (B), GSTP1 (C), GSTM1 (D), FN1 (E) and SERPINB9 (F) were measured in the CPA/EE+Met and GLP-1RA+Met groups at baseline and post treatment. n = 27–30. * P value < 0.05, ** P value < 0.01, *** P value < 0.001, paired t test