Epithelial Cell Line Derived from Endometriotic Lesion Mimics Macrophage Nervous Mechanism of Pain Generation on Proteome and Metabolome Levels.

Benjamin Neuditschko,Samuel M Meier-Menches,Marlene C Gerner,Christopher Gerner,Iveta Yotova,Lukas Skos,Marlene Leibetseder,Julia Brunmair,Gerhard Hagn

doi:10.3390/biom11081230

Benjamin Neuditschko, Samuel M Meier-Menches + Show 7 more

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https://doi.org/10.3390/biom11081230

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Abstract

Endometriosis is a benign disease affecting one in ten women of reproductive age worldwide. Although the pain level is not correlated to the extent of the disease, it is still one of the cardinal symptoms strongly affecting the patients’ quality of life. Yet, a molecular mechanism of this pathology, including the formation of pain, remains to be defined. Recent studies have indicated a close interaction between newly generated nerve cells and macrophages, leading to neurogenic inflammation in the pelvic area. In this context, the responsiveness of an endometriotic cell culture model was characterized upon inflammatory stimulation by employing a multi-omics approach, including proteomics, metabolomics and eicosanoid analysis. Differential proteomic profiling of the 12-Z endometriotic cell line treated with TNFα and IL1β unexpectedly showed that the inflammatory stimulation was able to induce a protein signature associated with neuroangiogenesis, specifically including neuropilins (NRP1/2). Untargeted metabolomic profiling in the same setup further revealed that the endometriotic cells were capable of the autonomous production of 7,8-dihydrobiopterin (BH2), 7,8-dihydroneopterin, normetanephrine and epinephrine. These metabolites are related to the development of neuropathic pain and the former three were found up-regulated upon inflammatory stimulation. Additionally, 12-Z cells were found to secrete the mono-oxygenated oxylipin 16-HETE, a known inhibitor of neutrophil aggregation and adhesion. Thus, inflammatory stimulation of endometriotic 12-Z cells led to specific protein and metabolite expression changes suggesting a direct involvement of these epithelial-like cells in endometriosis pain development.

Highlights

Endometriosis is a chronic inflammatory disease describing the abnormal growth of uterine tissue outside of the uterine cavity in the pelvic area [1]
An in-vitro model of endometriosis was investigated to evaluate the effects of such inflammatory stimuli on endometriotic cells by means of a multi-omics approach, including untargeted shotgun proteomics, metabolomics and eicosanoid analysis (Figure S1)
Eicosanoid Analysis Reveals the Uptake of Eicosanoid Precursors by 12-Z Cells from the Eicosanoids from the supernatants of control and inflammatory stimulated 12-Z cells were enriched by a solid-phase extraction protocol and subsequently analyzed by mass spectrometry

Summary

Introduction

Endometriosis is a chronic inflammatory disease describing the abnormal growth of uterine tissue outside of the uterine cavity in the pelvic area [1]. Endometriotic cells are characterized by invasive phenotypes. They successfully attach to pelvic organs and cause pelvic inflammation [2,3]. Studies estimate that the disease is affecting about 1 in 10 women worldwide. The clinical symptoms include dysmenorrhea, dyspareunia, chronic pelvic pain and infertility. There exists no curative treatment [1]. As one of Biomolecules 2021, 11, 1230.

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