Effect of ammonium tetrathiomolybdate on the inflammatory state and sensory reinnervation of endometriotic-like lesions induced in mice

CONFORTI R; DELSOUC M; ZABALA A; VALLCANERAS S; MONTT-GUEVARA M; GIANNINI A; SIMONCINI T; CASAIS M

Amsterdam

Congreso; 15th Congress of the European Society of Gynecology; 2023

European Society of Gynecology

Context: Endometriosis (EDT) is a chronic, inflammatory, and highly prevalent gynecological disease. Pain is one of the main symptoms that greatly affect the life quality of patients. We have shown that ammonium tetrathiomolybdate (TM) inhibits the growth of endometriotic-like lesions in mice with a good safety profile. Since several studies suggest a relationship between inflammation and aberrant innervation with pain in EDT, it would be interesting to analyze possible changes in these processes as a consequence of TM administration. Objective: Our objective was to analyze whether oral administration of TM to EDT-induced mice alters the levels of proinflammatory cytokines and the sensory reinnervation of lesions. Methods: Female C57BL/6 mice were divided into three groups: 1) Sham (placebo surgery), 2) EDT, and 3) EDT+TM. The EDT induction consisted of autologous uterine tissue transplantation to the intestinal mesentery. TM was administered orally (0.30 mg/day/mouse) from postoperative day 15, and animals were euthanized one month after inducing pathology. Peritoneal fluid (PF) was collected to determine the levels of the cytokines TNF-α and IL-1β by ELISA. Lesions were processed for the analysis of the brain-derived neurotrophic factor (BDNF) and neural growth factor (NGF) protein expression by ELISA, and mRNA expression of Tnfa, Il1b, Bdnf, Ngf, p75 neurotrophin receptors (p75NTR), Pgp9.5 (a pan-neuronal marker), tachykinin 1 (Tac1), tachykinin receptor 1 (Tacr1), calcitonin-gene related peptide (Cgrp), calcitonin receptor-like receptor (Crlr), receptor activity-modifying protein 1 (Ramp1), and transient receptor potential cation channel V1 (Trpv1) by RT-qPCR. Results: TM administration decreased the levels of both proinflammatory cytokines in PF and their mRNA expression in lesions. Furthermore, the treatment decreased the expression of Bdnf and Ngf, involved in neuron survival and differentiation, and their common receptor p75NTR in the lesions. It even reduced the NGF protein expression. In the lesions, TM also decreased the mRNA expression of Pgp9.5, the sensory neuropeptides Tac1 and Cgrp, and the components required to form functional CGRP receptors (Crlr and Ramp1), compared to the EDT group. Even the treatment decreased the Trpv1 mRNA expression, which is associated with chronic pelvic pain in EDT. Conclusions: Our results suggest that TM decreases the levels of the main cytokines involved in EDT and affects the sensory reinnervation of endometriotic-like lesions induced in mice.