Reduced fertility in an adenomyosis mouse model is associated with an altered immune profile in the uterus during the implantation period
Authors: Bourdon M , Maget A S , Jeljeli M , Doridot L , Marcellin L , Thomas M , Chêne C , Chouzenoux S , Batteux F , Chapron C , Santullli P 
Published: November 2023

Abstract

STUDY QUESTION
Does a reduction in fertility and/or systemic immune cell change occur during the early implantation period in a mouse model of adenomyosis?

SUMMARY ANSWER
A reduction in fertility was observed in mice with adenomyosis, coinciding with local and systemic immune changes observed during the implantation period.

WHAT IS KNOWN ALREADY
Adenomyosis is a pathology responsible for impaired fertility in humans, with a still unclear pathophysiology. One hypothesis is that changes in immune cells observed in adenomyosis-affected uteri may alter fertility, notably the physiological immune environment necessary for successful implantation and a healthy pregnancy.

STUDY DESIGN, SIZE, DURATION
Randomly selected CD-1 female neonatal pups were orally dosed by administration of tamoxifen to induce adenomyosis (TAM group), while others received solvent only (control group). From 6 weeks of life, CD-1 mice of both groups were mated to study impaired fertility and related local and/or systemic immune cell changes during the early implantation period.

PARTICIPANTS/MATERIALS, SETTINGS, METHODS
To evaluate fertility and pregnancy outcomes, ultrasound imaging was performed at E (embryonic day) 7.5 and E11.5 to count the number of gestational sacs and the number of resorptions in eight mice of the TAM group and 16 mice of the control group. The mice were sacrificed at E18.5, and morphometric, functional (quantitative reverse transcription PCR; RT-qPCR), and histological analyses were performed on the placentas. To identify local and/or systemic immune changes during the early implantation period, 8 mice of the TAM group and 12 mice of the control group were sacrificed at E4.5. Uterine horns and spleens were collected for flow cytometry and RT-qPCR analyses to study the immune cell populations. To investigate the profile of the cytokines secreted during the early implantation period at the systemic level, supernatants from stimulated spleen cells were analyzed by multiplex immunoassay analysis.

MAIN RESULTS AND THE ROLE OF CHANCE
By ultrasound imaging, we observed a lower number of implantation sites (P &lt; 0.005) and a higher number of resorptions (P &lt; 0.001) in the TAM group, leading to smaller litters (average number of fetuses per litter: 1.00 [0.00; 5.25] in the TAM group versus 12.00 [9.50; 13.75] in the control group (P &lt; 0.001). Histological and morphometric analyses of the placentas at E18.5 showed a higher junctional/labyrinthine area ratio in the TAM group (P = 0.005). The expression levels of genes that play a role in vascularization and placental growth (Vegf (P &lt; 0.001), Plgf (P &lt; 0.005), Pecam (P &lt; 0.0001), and Igf2 (P = 0.002)) were reduced in the TAM group. In the TAM group, the percentages of macrophages, natural killer (NK) cells, and dendritic cells (DC) were significantly decreased in the uterus around the implantation period. However, the number of M1 macrophages was increased. Both macrophages and DC had an increased activation profile (higher expression of MCHII, P = 0.012; CD80, P = 0.015; CCR7, P = 0.043 for macrophages, and higher expression of CD206, P = 0.018; CXCR4, P = 0.010; CCR7, P = 0.006, MCHII, P = 0.010; and CD80, P = 0.012 for DC). In spleen, an increase in the activation of macrophages (CCR7, P = 0.002; MCHII, P = 0.001; and CD80, P = 0.034) and DC was observed in the TAM group (CCR7, P = 0.001; MCHII, P = 0.001; Ly6C, P = 0.015). In the uteri and the spleen, we observed increased percentages of CD4+ T lymphocytes (P = 0.0237 and P = 0.0136, respectively) in the TAM group and, in the uteri, an increased number of regulatory T cells (P = 0.036) compared with the controls.

LARGE SCALE DATA
Not applicable.

LIMITATIONS, REASONS FOR CAUTION
This study is limited by the use of an animal model and the lack of intervention.

WIDER IMPLICATIONS OF THE FINDINGS
These data support involvement of innate and adaptive immune cells in the implantation failure and the increased rate of resorption observed in the mouse model of adenomyosis. This substantiates the need for additional research in this domain, with the goal of addressing fertility challenges in women affected by this condition.

STUDY FUNDING/COMPETING INTEREST(S)
None.