MODEL FOR GLUCOSE INDUCED BASEMENT MEMBRANE THICKENING

葡萄糖引起的基底膜增厚模型

• 批准号: 2734150
• 负责人: MICHAEL Peter SARRAS
• 金额: $ 19.45万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-07-01 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2734150
• 关键词: Cnidaria; SDS polyacrylamide gel electrophoresis; alternatives to animals in research; basement membrane; cell cell interaction; cellular pathology; collagen; enzyme activity; epithelium; extracellular matrix; fibronectins; genetic library; genetic translation; immunocytochemistry; laminin; medical complication; membrane biogenesis; membrane structure; messenger RNA; metalloendopeptidases; model; molecular pathology; monoclonal antibody; nucleic acid probes; polymerization; western blottings

A thickening in basement membranes (or epithelial-associated extracellular matrix (ECM)) is a common feature of a number of disease states. For example, ECM thickening has been observed 1) associated with the renal tubules and glomeruli (GBM) of patients with polycystic kidney disease (PKD), 2) associated with the seminiferous tubules in male patients with certain forms of impaired fertility, 3) associated with the GBM of patients with Alport syndrome, and 4) in patients with diabetes mellitus. Progressive secondary complications of these diseases have been tied to abnormalities associated with this ECM thickening. Although an explanation as to the mechanisms by which abnormal ECM thickening occurs is not currently available, it is evident that the normal biosynthesis and maintenance of ECM is an important aspect of healthy differentiated tissues. As indicated, the cellular and molecular basis for alterations in ECM formation is not understood, but may be explained in terms of problems in the balance of ECM component synthesis, polymerization, and turnover. These processes all involve cell/ECM interactions to some degree and imply that ECM formation is normally tightly controlled by both cellular and extracellular processes. Experimental approaches to evaluate the mechanisms which govern normal ECM formation and the mechanisms by which abnormally thickened ECM occurs have been hampered by a lack of in vitro and in vivo cellular models. In order to approach these problems we have developed an in vivo cellular model in which epithelial-associated ECM formation can be experimentally induced in a short time frame for subsequent analysis of the cellular mechanisms involved in the process. In addition, methods have been developed to trigger the formation of abnormally thickened ECM so that both the normal and abnormal process can be directly compared and evaluated. The model we have developed is simply comprised of an epithelial bilayer with an intervening ECM. In addition, we have determined that this in vivo model responds to hyperglycemic conditions by thickening its ECM as observed in various pathological conditions. As opposed to vertebrate animal models currently available however, the cell system we utilize develops an ECM within 24-96 hr and doubles the thickness of this ECM within this same time frame when exposed to elevated levels of glucose. This model was developed using the Cnidarian, Hydra vulgaris. The proposed project will utilize this in vivo model to analyze normal ECM formation and determine if abnormal thickening of ECM results from 1) cellular abnormalities in the synthesis and accumulation of ECM components, 2) abnormalities in the extracellular assembly of ECM components., and/or 3) abnormalities in the degradation of ECM components. A combination of morphological, biochemical, and molecular approaches will be utilized to test these hypotheses. This project will provide basic information on the cellular mechanisms of ECM formation under normal conditions and under conditions in which ECM thickening occurs.

基底膜（或上皮相关细胞外膜）增厚 矩阵（ECM））是许多疾病状态的共同特征。 为了 例如，已观察到 ECM 增厚 1) 与肾功能相关 多囊肾病患者的肾小管和肾小球 (GBM) (PKD), 2) 与男性患者的曲细精管相关 某些形式的生育能力受损，3) 与 GBM 相关 阿尔波特综合征患者，4) 糖尿病患者。 这些疾病的进行性继发并发症与 与这种 ECM 增厚相关的异常。 虽然一个 解释异常 ECM 增厚发生的机制 目前还没有，很明显正常的生物合成和 维持ECM是健康分化的一个重要方面 组织。 如前所述，改变的细胞和分子基础 ECM 的形成尚不清楚，但可以用以下方式解释： ECM组分合成、聚合和平衡的问题 周转。 这些过程都涉及细胞/ECM 相互作用 程度并意味着 ECM 的形成通常受到两者的严格控制 细胞和细胞外过程。 评估实验方法 控制正常 ECM 形成的机制以及通过 异常增厚的 ECM 的发生受到缺乏 体外和体内细胞模型。 为了解决这些问题我们 开发了一种体内细胞模型，其中上皮相关 ECM 的形成可以在短时间内通过实验诱导 随后分析该过程中涉及的细胞机制。 此外，还开发了一些方法来触发形成 异常增厚的 ECM，以便正常和异常过程都可以 直接进行比较和评价。 我们开发的模型很简单 由上皮双层和插入的 ECM 组成。 此外， 我们已经确定该体内模型对高血糖有反应 如在各种病理中观察到的那样，通过增厚其 ECM 来改善病情 状况。 与目前可用的脊椎动物模型相反 然而，我们使用的细胞系统会在 24-96 小时内形成 ECM，并且 当暴露时，在同一时间范围内该 ECM 的厚度加倍 导致葡萄糖水平升高。 该模型是使用 刺胞动物、普通九头蛇。 拟议的项目将在体内利用这一点 分析正常 ECM 形成并确定是否异常增厚的模型 ECM 的结果来自 1) 细胞合成异常和 ECM成分积累，2）细胞外异常 ECM 组件的组装。和/或 3) 降解异常 ECM 组件。 形态学、生化学和 将利用分子方法来检验这些假设。 这 项目将提供ECM细胞机制的基本信息 在正常条件下和 ECM 条件下形成 发生增厚。

项目成果

Identification and characterization of hydra metalloproteinase 2 (HMP2): a meprin-like astacin metalloproteinase that functions in foot morphogenesis.

水螅金属蛋白酶 2 (HMP2) 的鉴定和表征：一种类似 meprin 的虾蛋白金属蛋白酶，在足部形态发生中发挥作用。

• DOI: 10.1242/dev.127.1.129
• 发表时间: 2000
• 期刊: Development (Cambridge, England)
• 作者: Yan,L;Fei,K;Zhang,J;Dexter,S;SarrasJr,MP
• 通讯作者: SarrasJr,MP

A cnidarian homologue of translationally controlled tumor protein (P23/TCTP).

翻译控制肿瘤蛋白 (P23/TCTP) 的刺胞动物同源物。

• DOI: 10.1007/s004270000088
• 发表时间: 2000
• 期刊: Development genes and evolution
• 影响因子: 2.4
• 作者: Yan,L;Fei,K;Bridge,D;SarrasJr,MP
• 通讯作者: SarrasJr,MP

Molecular and biological characterization of a zonula occludens-1 homologue in Hydra vulgaris, named HZO-1.

寻常水螅中的 zonula occlusionns-1 同源物（名为 HZO-1）的分子和生物学特征。

• DOI: 10.1007/s004270000103
• 发表时间: 2000
• 期刊: Development genes and evolution.
• 作者: Fei,K;Yan,L;Zhang,J;SarrasJr,MP
• 通讯作者: SarrasJr,MP

Molecular, biochemical and functional analysis of a novel and developmentally important fibrillar collagen (Hcol-I) in hydra.

对水螅中一种新型且对发育重要的纤维状胶原蛋白 (Hcol-I) 进行分子、生化和功能分析。

• DOI: 10.1242/dev.127.21.4669
• 发表时间: 2000
• 期刊: Development (Cambridge, England)
• 作者: Deutzmann,R;Fowler,S;Zhang,X;Boone,K;Dexter,S;Boot-Handford,RP;Rachel,R;SarrasJr,MP
• 通讯作者: SarrasJr,MP

BMP-1 and the astacin family of metalloproteinases: a potential link between the extracellular matrix, growth factors and pattern formation.

• DOI: 10.1002/bies.950180604
• 发表时间: 1996-06
• 期刊: BioEssays : news and reviews in molecular, cellular and developmental biology
• 作者: M. Sarras