1. Effect of GnRH-agonist downregulation on serum AMH levels: a prospective cohort study with repeated measurements
P. Drakopoulos1, A. van de Vijver1, J. Parra2, E. Anckaert1, J. Schiettecatte1, J. Smitz1, C. Blockeel1, M. Hund3, W. D. J. Verhagen- Kamerbeek3, Y. He4, H. Tournaye1, N. P. polyzos5,6,7
1Centre for Reproductive Medicine, Universitair Ziekenhuis Brussel, Vrije Universiteit Brussel, Brussels, Belgium; 2Department of Statistics, Universitair Ziekenhuis Brussel, Vrije Universiteit Brussel, Brussels, Belgium; 3Roche Diagnostics International Ltd, Rotkreuz, Switzerland; 4Roche Diagnostics GmbH, Penzberg, Germany; 5Department of Reproductive Medicine, Dexeus University Hospital, Barcelona Spain; 6Faculty of Medicine and Pharmacy, Vrije Universiteit Brussel, Brussels, Belgium; 7Faculty of Health, Aarhus University, Aarhus, Denmark

2. Study design
Prospective cohort study performed at a center for reproductive medicine in a tertiary university hospital (July 2015 – March 2016)
Primary endpoint: effect of a GnRH-agonist on serum AMH concentration after 7 and 14 days of treatment.
†Baseline measurement taken before treatment was initiated
SC, subcutaneous; UZB, Universitair Ziekenhuis Brussel

3. Patient demographics and baseline characteristics
All patients
(n=52)
Mean ± SD age, years
35.8 ± 3.6
Mean BMI ± SD, kg/m2
23.5 ± 3.6
Race, n (% of total study population)
White
Asian
Black
Other
48 (92.3)
1 (1.9)
2 (3.9)
Smoking status, n (% of total study population
Yes No
Missing
4 (7.7)
47 (90.4)
Mean ± SD AMH at baseline, ng/mL
1.65 ± 0.8
Mean AFC ± SD at baseline
11.3 ± 4.3
AFC class, n (% of total study population)
0–7
8–15
>15
6 (11.5)
35 (67.3)
5 (9.6)
AFC, antral follicle count; AMH, anti-Müllerian hormone; BMI, body mass index; SD, standard deviation

4. Box plots of serum AMH concentrations at baseline, and Days 7 and 14 of GnRH-agonist treatment (n=52)
Serum AMH concentrations followed a v-shape from baseline through Day 7 to 14
***P<0.001 versus baseline; Circles represent individual patient data; crosses are the mean value; horizontal lines summarize the median (within the box) and the first and third quartiles; whiskers are 1.5 x the interquartile range.
AMH, anti-Müllerian hormone

5. Median % change in concentration (95% CI) [P value*]
Percentage changes in serum concentrations of AMH, FSH, LH, E2 and progesterone (n=52)
Hormone
Median % change in concentration (95% CI) [P value*]
Day 7 vs baseline
Day 14 vs baseline
Day 14 vs Day 7
AMH
–14.9 (–23.0, –10.1)
[<0.05]
17.4 (10.4, 35.9)
[<0.001]
48.3 (34.7, 57.4)
FSH
–19.8 (–27.5, 3.04)
[NS]
–7.31 (–25.0, 22.6)
5.47 (–6.97, 22.6) LH
–9.96 (–17.6, 17.2)
–43.2 (–55.4, –33.1)
–47.6 (–52.2, –41.0) E2
–28.6 (–60.6, 25.3)
–94.8 (–95.9, –93.4)
–90.5 (–94.5, –81.0)
Progesterone
–69.2 (–85.5, 0.168)
–95.8 (–97.7, –94.5)
–90.9 (–94.7, –79.1)
Significant decreases in serum LH, E2 and progesterone at Day 14 confirm the ability of GnRH-agonist treatment to downregulate pituitary function
*Wilcoxon signed rank test, median of the percentage change Statistical significance for AMH was defined as P<0.05; for other hormones P< (Bonferroni correction)
AMH, anti-Müllerian hormone; E2, estradiol; FSH, follicle stimulating hormone; LH, luteinizing hormone; NS, not significant

6. Individual serum AMH concentrations for the four aliquots measured for each patient per visit: baseline, and Days 7 and 14 (n=52) 4 3 2 1 BL 7 14
Serum AMH (ng/mL) 5 6 9 10 11 12 13 15 16 17 18 19 20 21 22 23 24 26 27 28 29 30 31 32 33 34 35 36 37 40 41 43 44 45 46 47 48 49 50 51 52 53 54 57
SJ1
SJ2
Elecsys® AMH assay demonstrated excellent precision across the four aliquots measured for each serum sample
Circles represent the four aliquots for each sample at each visit BL, baseline

7. Conclusions
GnRH-agonist treatment had a statistically significant effect on serum AMH concentrations in women with infertility who were planning to undergo ovarian stimulation.
In a clinical setting, these changes may lead to an inaccurate prediction of ovarian reserve.
Serum AMH should therefore be measured before starting GnRH- agonist treatment to allow for reliable assessment of ovarian reserve and prediction of response to ovarian stimulation.