Williams Katherine E, Miroshnychenko Olga, Johansen Eric B, Niles Richard K, Sundaram Rajeshwari, Kannan Kurunthachalam, Albertolle Matthew, Zhou Yan, Prasad Namrata, Drake Penelope M, Giudice Linda C, Hall Steven C, Witkowska H Ewa, Buck Louis Germaine M, Fisher Susan J
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.
Sandler-Moore Mass Spectrometry Core Facility, University of California San Francisco, San Francisco, CA, USA.
J Proteomics. 2015 Jan 15;113:194-205. doi: 10.1016/j.jprot.2014.09.015. Epub 2014 Oct 13.
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.
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.
子宫内膜异位症是子宫内膜的异位生长，影响6%至11%的育龄妇女，与盆腔疼痛和不孕有关。我们研究了患有子宫内膜异位症的女性与未发现手术可见子宫内膜异位症的个体的腹腔液（PF）、尿液和网膜脂肪（OF）蛋白质组。所有参与者均参加了由美国国立儿童健康与人类发展研究所资助的子宫内膜异位症研究。进行了两步蛋白质组学研究。第一步是广泛调查，采用半定量凝胶液相色谱-质谱（MS）工作流程：SDS-PAGE分级分离、胰蛋白酶消化和LC-MS/MS。结果显示样本完整性，但未检测到患有和未患有子宫内膜异位症的女性之间存在任何差异。第二步是对OF样本进行定量分析。我们使用了另一组来自患或未患疾病女性的样本（n = 30），并采用等压质量标签（iTRAQ）化学方法标记肽段，通过二维液相色谱-质谱联用技术进行蛋白质鉴定和定量。三种蛋白质——基质金属蛋白酶-9、中性粒细胞弹性蛋白酶和FAM49B——在患有子宫内膜异位症的女性样本中的丰度显著降低。有趣的是，中性粒细胞弹性蛋白酶和FAM49B的水平与先前在相同样本中测量的一部分内分泌干扰化学物质（EDC）的较高水平相关。这些实验结果表明了将子宫内膜异位症与OF蛋白质组变化和EDC水平相关联的可行性。
子宫内膜异位症是子宫内膜的病理性生长，与包括疼痛和不孕在内的重大疾病相关。然而，这种常见病症的病因尚不清楚。本研究确定子宫内膜异位症是否与腹腔液、尿液和/或网膜脂肪的蛋白质组成变化有关。在患有与未患有子宫内膜异位症的女性的OF样本中，一种功能未知的蛋白质（FAM49B）和两种蛋白酶（基质金属蛋白酶-9、中性粒细胞弹性蛋白酶）表达下调。这些发现表明在远离子宫内膜异位病变的部位存在蛋白酶失衡。此外，FAM49B和中性粒细胞弹性蛋白酶水平与在相同样本中定量的一部分环境化学物质的较高水平相关，提示了其他可能的关联。因此，这项工作产生了将在进一步研究中进行检验的假设。
