ENDOMETRIOTIC HAPTOGLOBIN ALTERS MACROPHAGE FUNCTION

子宫内膜异位触珠蛋白改变巨噬细胞功能

• 批准号: 6616456
• 负责人: KATHY L TIMMS
• 金额: $ 30.6万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6616456
• 关键词: aerobiosis; blood lipoprotein biosynthesis; cellular pathology; chemotaxis; clinical research; cytokine; endometriosis; endometrium; enzyme linked immunosorbent assay; female; growth factor; haptoglobins; human subject; immunoregulation; integrins; leukocyte activation /transformation; macrophage; method development; molecular pathology; phagocytosis; polymerase chain reaction; protein structure function; secretory protein; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): Endometriosis affects 5 1/2 million reproductive age women and girls in the USA and Canada, and millions more worldwide, causing pelvic pain and infertility. Diagnosis and treatment require costly, invasive surgery to identify and ablate ectopic endometrial tissue. Endometriosis is one of the three top reasons for hysterectomy in the USA; over 1/2 million hysterectomies are performed annually at an estimated cost of more than $5 billion. Yet, the pathogenesis of endometriosis remains poorly defined. The long-term objectives of this research are to develop novel methods of medical management by characterizing endometriotic secretory proteins that correlate with the cellular and molecular pathogenic mechanisms of endometriosis. This research evolves from the discovery that endometriotic lesions actually synthesize and secrete haptoglobin (Hp). Intriguingly, endometriotic haptoglobin (eHp) is differentially glycosylated compared to hepatic Hp. Preliminary data support a pathologically relevant role for eHp in the aberrant immunological phenomena that support the disease process in women with endometriosis. The hypothesis to be tested is that by expressing eHp, endometriotic tissues from women with endometriosis avoid phagocytic eradication while stimulating peritoneal macrophage inflammatory cytokine secretion. In turn, the macrophage cytokines increase endometriotic tissue eHp production, creating a local, feed-forward loop between ectopic endometrium and macrophages favoring the establishment of endometriosis. To test this hypothesis, peritoneal macrophages, and endometriotic lesions when present, will be collected from women without and with endometriosis. These immune cells and tissues will be used to investigate three specific aims: 1) Identify the effects of eHp on peritoneal macrophage phagocytosis by analyzing the five steps of macrophage function in vitro including chemotaxis, adherence, ingestion, oxidative metabolism and activation. 2) Characterize a ligand/receptor mechanism whereby eHp causes aberrant macrophage function, by selectively altering eHp glycans and/or blocking peritoneal macrophage integrins. 3) Quantify the effects of macrophage inflammatory cytokines and growth factors on eHp synthesis and secretion. These experiments will provide insight into the pathogenesis of endometriosis by determining if endometriotic tissues, peritoneal macrophages or both are responsible for this pathology, if this mechanism is unique to women with endometriosis and confirm our feedforward hypothesis. As a result, novel non-invasive strategies for early detection and innovative treatment of endometriosis may be developed that markedly reduce the health burden of this malady.

描述（由申请人提供）：子宫内膜异位症影响美国和加拿大的550万育龄妇女和女孩，以及全球数百万人，引起盆腔疼痛和不孕症。诊断和治疗需要昂贵的侵入性手术来识别和消融异位子宫内膜组织。子宫内膜异位症是美国子宫切除术的三大原因之一;每年进行超过1/5百万例子宫内膜切除术，估计费用超过50亿美元。然而，子宫内膜异位症的发病机制仍然不清楚。本研究的长期目标是通过表征与子宫内膜异位症的细胞和分子致病机制相关的子宫内膜分泌蛋白来开发新的医疗管理方法。这项研究是从发现幽门螺杆菌病变实际上合成和分泌触珠蛋白（Hp）。有趣的是，与肝Hp相比，嗜酸性触珠蛋白（eHp）的糖基化差异。初步数据支持eHp在异常免疫现象中的病理相关作用，这些异常免疫现象支持子宫内膜异位症妇女的疾病过程。有待检验的假设是，通过表达eHp，子宫内膜异位症妇女的增生组织避免了吞噬细胞的根除，同时刺激腹腔巨噬细胞的炎性细胞因子分泌。反过来，巨噬细胞细胞因子增加异位组织eHp的产生，在异位子宫内膜和巨噬细胞之间形成有利于建立子宫内膜异位症的局部前馈回路。 为了检验这一假设，将从患有和不患有子宫内膜异位症的女性中收集腹膜巨噬细胞和增生性病变（如果存在）。这些免疫细胞和组织将被用于研究三个特定的目的：1）通过分析体外巨噬细胞功能的五个步骤（趋化、粘附、摄取、氧化代谢和活化）来确定eHp对腹腔巨噬细胞吞噬功能的影响。2)通过选择性改变eHp聚糖和/或阻断腹膜巨噬细胞整合素，表征eHp导致巨噬细胞功能异常的配体/受体机制。3)定量巨噬细胞炎性细胞因子和生长因子对eHp合成和分泌的影响。这些实验将通过确定子宫内膜异位症组织、腹腔巨噬细胞或两者是否对这种病理负责来深入了解子宫内膜异位症的发病机制，如果这种机制是子宫内膜异位症女性独有的，并证实我们的前馈假设。因此，新的非侵入性策略，早期发现和创新的治疗子宫内膜异位症可能会开发，显着减少这种疾病的健康负担。