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: 6721471
• 负责人: KATHY L TIMMS
• 金额: $ 31.82万
• 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/6721471
• 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(申请人提供)：子宫内膜异位症在美国和加拿大影响550万育龄妇女和女孩，在全球范围内影响数百万人，导致盆腔疼痛和不孕不育。诊断和治疗需要昂贵的侵入性手术来识别和消融异位内膜组织。子宫内膜异位症是美国子宫切除术的三大原因之一；每年有超过150万例子宫切除术，估计成本超过50亿美元。然而，子宫内膜异位症的发病机制仍不明确。这项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发病机制相关的子宫内膜异位症分泌蛋白来开发新的医疗管理方法。这项研究源于子宫内膜异位症病变实际上合成和分泌结合珠蛋白(HP)的发现。有趣的是，子宫内膜异位结合珠蛋白(EHP)与肝性HP相比糖基化程度不同。初步数据支持EHP在支持子宫内膜异位症妇女疾病进程的异常免疫现象中的病理相关作用。需要检验的假设是，子宫内膜异位症患者的子宫内膜异位症组织通过表达EHP，避免了吞噬细胞的清除，同时刺激了腹膜巨噬细胞炎症细胞因子的分泌。反过来，巨噬细胞因子增加了子宫内膜异位症组织EHP的产生，在异位内膜和巨噬细胞之间建立了一个局部的前馈环路，有利于子宫内膜异位症的建立。 为了验证这一假设，将从没有和有子宫内膜异位症的妇女身上收集腹膜巨噬细胞和子宫内膜异位症病变。这些免疫细胞和组织将被用来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，包括趋化、黏附、吞噬、氧化代谢和激活，确定EHP对巨噬细胞吞噬功能的影响。2)研究EHP通过选择性改变EHP多糖和/或阻断腹膜巨噬细胞整合素，导致巨噬细胞功能异常的配体/受体机制。3)定量检测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是子宫内膜异位症组织、腹膜巨噬细胞还是两者共同导致这一病理机制，从而深入了解子宫内膜异位症的发病机制，并证实我们的前馈假说。因此，早期发现和创新治疗子宫内膜异位症的新的非侵入性策略可能会被开发出来，从而显著减轻这种疾病的健康负担。