ENDOMETRIOSIS-ASSOCIATED SECRETORY PROTEINS

子宫内膜异位症相关分泌蛋白

• 批准号: 2201537
• 负责人: KATHY L TIMMS
• 金额: $ 10.01万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-04-01 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2201537
• 关键词: antiserum; biomarker; complementary DNA; diagnosis design /evaluation; endometriosis; endometrium; enzyme linked immunosorbent assay; female; fertility; gene expression; genetic library; human genetic material tag; human subject; human tissue; immunocytochemistry; laboratory rabbit; laboratory rat; menstrual cycle; molecular cloning; northern blottings; nucleic acid sequence; pregnancy; progesterone; protein purification; protein sequence; protein structure function; radioimmunoassay; reproductive system disorder diagnosis; secretory protein; tissue /cell culture; western blottings

Endometriosis is a disease that causes debilitating pain and loss of fertility in women. Despite these devastating effects, the mechanism by which this enigmatic disease exerts these pathogenicities is unknown. Although histologically similar to uterine endometrium, endometriotic tissue is biochemically different from its uterine counterpart in a variety of fashions. The altered biochemical characteristics of the endometriotic tissue may be involved with anomalies of the immune system, ovulation, fertilization, and implantation which have all been implicated as causes of endometriosis-associated infertility. Using a rat model for endometriosis, two previously unknown endometriosis-associated proteins have been identified. A progesterone- induced uterine protein, PUP-1, may play a role in normal reproductive processes including implantation. Curiously, synthesis and/or secretion of PUP-1 by the endometriotic implant is 24 hours out of phase with that of the uterine endometrium. A second protein, EP-1, is synthesized by endometriotic but not endometrial tissue. EP-1 may be a useful marker of endometriosis and have a role in the pathophysiology of the disease. PUP-1 and EP-1 have recently been identified in culture media from purified rat and human endometrial and endometriotic implant stromal cells, respectively. The goal of this proposal is to purify and characterize both rat and human PUP-1 and EP-1 in order to obtain insight into the structure, mode of expression and physiological function of these proteins. The specific aims of this project are to: 1) develop a purification scheme for PUP-1 and EP-1; 2) generate a) antisera, b) radioimmunoassays and c) limited protein sequence data for PUP-1 and EP-1; and 3) molecularly clone cDNA representing PUP-1 and EP-1 and obtain nucleotide sequence data. Protein purification will employ cell culture and chromatographic techniques. the antisera, radioimmunoassays and protein sequence data derived from the purified proteins will be used to detect and monitor the distribution of PUP-1 and EP-1 in various tissues and in sera throughout the reproductive cycle, during early pregnancy and following steroid treatment as well as to obtain DNA sequence data for PUP-1 and EP-1. The nucleic acid sequence data will be compared to other sequence information in data banks to provide further insight into the identity and possible function of the proteins. These studies will support long term goals which include determining the role of PUP-1 and EP-1 in reproductive function and the pathophysiology of endometriosis. The characterization of PUP-1 and EP-1 may also lead to the development of non-invasive serum markers for progesterone- dependent endometrial function and the disease endometriosis. Ultimately, these studies may lead to improved diagnostic, prognostic and therapeutic methods for the management of endometriosis.

子宫内膜异位症是一种导致衰弱性疼痛和子宫内膜异位症的疾病 女性的生育能力。 尽管有这些破坏性影响，但该机制 这种神秘的疾病究竟是由何种疾病产生这些致病性的，目前尚不清楚。 尽管组织学上与子宫内膜相似，但子宫内膜异位症 组织在生化上与子宫对应物不同 各种时尚。 生化特性的改变 子宫内膜异位组织可能与免疫系统异常有关， 排卵、受精和着床都受到影响 作为子宫内膜异位症相关不孕的原因。 使用子宫内膜异位症大鼠模型，两种以前未知的 子宫内膜异位症相关蛋白已被鉴定。 黄体酮—— 诱导子宫蛋白 PUP-1 可能在正常生殖中发挥作用 过程包括植入。 奇怪的是，合成和/或分泌 子宫内膜异位植入物产生的 PUP-1 与子宫内膜异位植入物产生的 PUP-1 的相位相差 24 小时 子宫内膜。 第二种蛋白质 EP-1 是由以下物质合成的 子宫内膜异位但不是子宫内膜组织。 EP-1 可能是一个有用的标记 子宫内膜异位症的发生并在该疾病的病理生理学中发挥作用。 最近在以下来源的培养基中发现了 PUP-1 和 EP-1： 纯化的大鼠和人子宫内膜和子宫内膜异位植入基质 细胞，分别。 该提案的目标是纯化和表征大鼠和 人类 PUP-1 和 EP-1 以深入了解其结构、模式 这些蛋白质的表达和生理功能。 具体的 该项目的目标是：1）开发PUP-1的纯化方案 和 EP-1； 2) 生成 a) 抗血清、b) 放射免疫测定和 c) 有限 PUP-1 和 EP-1 的蛋白质序列数据； 3) 分子克隆 cDNA 代表PUP-1和EP-1并获得核苷酸序列数据。 蛋白质纯化将采用细胞培养和色谱法 技术。 抗血清、放射免疫测定和蛋白质序列数据 从纯化的蛋白质中提取的蛋白质将用于检测和监测 PUP-1 和 EP-1 在各种组织和血清中的分布 生殖周期、怀孕早期和使用类固醇后 处理以及获得 PUP-1 和 EP-1 的 DNA 序列数据。 这 核酸序列数据将与其他序列信息进行比较 存储在数据库中，以进一步了解身份和可能的情况 蛋白质的功能。 这些研究将支持长期目标，其中包括确定 PUP-1 和 EP-1 在生殖功能和病理生理学中的作用 子宫内膜异位症。 PUP-1 和 EP-1 的表征也可能导致 开发黄体酮非侵入性血清标记物 依赖子宫内膜功能和子宫内膜异位症。 最终，这些研究可能会改善诊断、预后和治疗 治疗子宫内膜异位症的治疗方法。