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Abstract
BackgroundEndometriosis is a chronic and underdiagnosed disease which affects 5–10% of women of childbearing age and is characterized by growth of endometrial tissue outside of the uterus, most often in the peritoneal cavity. Delay in diagnosis is a major problem for management of this disorder, and treatment is often not initiated until the disease has progressed for many years. Although the exact etiology of endometriosis remains unknown, retrograde menstruation is recognized as a common underlying factor leading to the deposit of menstrual effluent (ME) into the peritoneal cavity. Differences in the cellular biology and genetics of the cells within ME are therefore likely to explain why endometriosis develops in only a subset of women.MethodsPatients with and without endometriosis were consented to provide ME. ME was analyzed by flow cytometry for CD45- and CD45+ cell populations or used to isolate stromal fibroblast cells. ME-derived stromal fibroblast cells were assessed using decidualization assays following the addition of cAMP and IGFBP-1 concentrations in the culture supernatants were measured by ELISA. In addition, RNA was collected and analyzed by RNA-Seq and qPCR for markers of decidualization and to identify differentially expressed genes in ME-derived stromal fibroblast cells obtained from controls and subjects with endometriosis (±cAMP).ResultsFlow cytometry analysis of cell subsets within the CD45+ fraction of ME revealed a significant decrease in the number of uterine NK cells in endometriosis patients compared with controls (p < 0.01). No other significant differences within either the CD45+ or CD45- cell populations were observed. Most strikingly, ME-derived stromal fibroblast cells cultured from endometriosis subjects showed impaired decidualization potential compared with controls. Highly significant differences in decidualization response were detected by measuring IGFBP-1 production at multiple time points after cAMP stimulation (p = 0.0025 at 6 h; p = 0.0045 at 24 h; p = 0.0125 at 48 h). RNA-Seq and qPCR analyses were used to identify genes differentially expressed by ME-derived stromal fibroblast cells obtained from endometriosis and control subjects.ConclusionsMenstrual effluent can be useful for investigating the pathobiology of endometriosis and for developing a non-invasive diagnostic for endometriosis which may lead to earlier and more effective treatments for this common disorder.
Highlights
Endometriosis is a chronic and underdiagnosed disease which affects 5–10% of women of childbearing age and is characterized by growth of endometrial tissue outside of the uterus, most often in the peritoneal cavity
Subject recruitment and enrollment Endometriosis subjects were recruited through the Research OutSmarts Endometriosis (ROSE) study and control subjects were recruited from the Genotype and Phenotype (GaP) registry; both studies were approved by the Institutional Review Board (IRB) of Northwell Health
menstrual effluent (ME) contains CD45+ and CD45- cell populations and ME collected from endometriosis subjects has fewer uterine natural killer (NK) cells Flow cytometric analysis was performed on fresh ME collected from control and endometriosis subjects for both CD45+ leukocyte populations and CD45- cell populations (Fig. 1a, b)
Summary
Endometriosis is a chronic and underdiagnosed disease which affects 5–10% of women of childbearing age and is characterized by growth of endometrial tissue outside of the uterus, most often in the peritoneal cavity. Endometriosis is a chronic disease characterized by the growth of endometrial-like tissue outside of the uterus It affects 5 to 10% of women globally (Giudice, 2010a). One of the most challenging problems of endometriosis is the prolonged time between disease onset and definitive diagnosis by laparoscopic surgery, which is approximately 7 to 10 years (Hadfield et al, 1996) This delay in diagnosis and early treatment is caused, in part, by the lack of non-invasive diagnostic tests for endometriosis (Ballard et al, 2006; Giudice, 2010b)
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