title: Urine, peritoneal fluid and omental fat proteomes of reproductive age women: Endometriosis-related changes and associations with endocrine disrupting chemicals.
abstract: Endometriosis, ectopic growth of the uterine lining (endometrium), which affects 6-11% of reproductive age women, is associated with pelvic pain and infertility. We investigated the peritoneal fluid (PF), urine and omental fat (OF) proteomes of women with endometriosis vs. individuals with no surgically visualized endometriosis. All participants were enrolled in the NICHD-funded ENDO Study. A two-step proteomic study was performed. The first, a broad survey, employed a semi-quantitative gel LC-mass spectrometry (MS) workflow: SDS PAGE fractionation, trypsin digestion and LC-MS/MS. The results showed sample integrity but failed to detect any differences between women with and without endometriosis. The second step was a quantitative analysis of OF samples. We employed another sample set (n=30) from women ± disease and isobaric mass-tag (iTRAQ) chemistry to label peptides and 2D LC-MS/MS for protein identification and quantification. Three proteins-matrix metalloproteinase-9, neutrophil elastase, and FAM49B-were significantly lower in abundance in samples from women with endometriosis. Interestingly, neutrophil elastase and FAM49B levels were associated with higher levels of a subset of endocrine disrupting chemicals (EDCs) that were previously measured in the same samples. The results of these experiments showed the feasibility of associating endometriosis with changes in the OF protein repertoire and EDC levels.
  
  
  CONCLUSIONS
  Endometriosis, pathological growth of the uterine lining, is associated with significant morbidities, including pain and infertility. However, the causes of this common condition are poorly understood. This study determined whether endometriosis was associated with changes in the protein composition of peritoneal fluid, urine and/or omental fat. A protein of unknown function (FAM49B) and two proteinases (metalloproteinase-9, neutrophil elastase) were down regulated in OF samples from women with versus without endometriosis. These findings suggested proteinase imbalances at sites that were distant from the endometriotic lesions. Additionally, FAM49B and neutrophil elastase levels were associated with higher levels of a subset of environmental chemicals that were quantified in the same samples, suggesting other possible associations. Thus, this work generated hypotheses that will be tested in further studies.
authors.affiliation: Sandler-Moore Mass Spectrometry Core Facility, University of California San Francisco, San Francisco, CA, USA; Center for Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA; Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Katherine E
authors.initials: KE
authors.lastname: Williams
authors.affiliation: Sandler-Moore Mass Spectrometry Core Facility, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Olga
authors.initials: O
authors.lastname: Miroshnychenko
authors.affiliation: Sandler-Moore Mass Spectrometry Core Facility, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Eric B
authors.initials: EB
authors.lastname: Johansen
authors.affiliation: Sandler-Moore Mass Spectrometry Core Facility, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Richard K
authors.initials: RK
authors.lastname: Niles
authors.affiliation: Division of Intramural Population Health Research, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Rajeshwari
authors.initials: R
authors.lastname: Sundaram
authors.affiliation: Wadsworth Center, New York State Department of Health, Empire State Plaza, Albany, NY, USA.
authors.forename: Kurunthachalam
authors.initials: K
authors.lastname: Kannan
authors.affiliation: Sandler-Moore Mass Spectrometry Core Facility, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Albertolle
authors.affiliation: Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA; The Eli & Edythe Broad Center for Regeneration Medicine and Stem Cell Research, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Sandler-Moore Mass Spectrometry Core Facility, University of California San Francisco, San Francisco, CA, USA; Center for Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA; Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Namrata
authors.initials: N
authors.lastname: Prasad
authors.affiliation: Sandler-Moore Mass Spectrometry Core Facility, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Penelope M
authors.initials: PM
authors.lastname: Drake
authors.affiliation: Center for Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA; Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Linda C
authors.initials: LC
authors.lastname: Giudice
authors.affiliation: Sandler-Moore Mass Spectrometry Core Facility, University of California San Francisco, San Francisco, CA, USA; Center for Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA; Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Steven C
authors.initials: SC
authors.lastname: Hall
authors.affiliation: Sandler-Moore Mass Spectrometry Core Facility, University of California San Francisco, San Francisco, CA, USA; Center for Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA; Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA.
authors.forename: H Ewa
authors.initials: HE
authors.lastname: Witkowska
authors.affiliation: Division of Intramural Population Health Research, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Germaine M
authors.initials: GM
authors.lastname: Buck Louis
authors.affiliation: Sandler-Moore Mass Spectrometry Core Facility, University of California San Francisco, San Francisco, CA, USA; Center for Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA; Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California San Francisco, San Francisco, CA, USA; Division of Maternal Fetal Medicine, University of California San Francisco, San Francisco, CA, USA; The Eli & Edythe Broad Center for Regeneration Medicine and Stem Cell Research, University of California San Francisco, San Francisco, CA, USA; Human Embryonic Stem Cell Program, University of California San Francisco, San Francisco, CA, USA; Department of Anatomy, University of California San Francisco, San Francisco, CA, USA. Electronic address: sfisher@cgl.ucsf.edu.
authors.forename: Susan J
authors.initials: SJ
authors.lastname: Fisher
mesh_terms: D000273:Adipose Tissue / Q000378:metabolism* / Q000473:pathology; D000328:Adult; D001202:Ascitic Fluid / Q000378:metabolism* / Q000473:pathology; D003278:Contraceptives, Oral, Hormonal / Q000008:administration & dosage*; D004715:Endometriosis / Q000473:pathology / Q000652:urine*; D005260:Female; D006801:Humans; D019272:Leukocyte Elastase / Q000652:urine; D020780:Matrix Metalloproteinase 9 / Q000652:urine; D009852:Omentum / Q000378:metabolism* / Q000473:pathology; D020543:Proteome / Q000378:metabolism*
chemical_list: D003278:Contraceptives, Oral, Hormonal; D020543:Proteome; D019272:Leukocyte Elastase; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9
keywords: Endometriosis; Environmental chemicals; Mass spectrometry; Omental fat; Peritoneal fluid; Urine