SUPPRESSING MACROPHAGE FUNCTIONS IN ATHEROSCLEROSIS

抑制动脉粥样硬化中的巨噬细胞功能

• 批准号: 2002455
• 负责人: Marko Z Radic
• 金额: $ 7.55万
• 依托单位: ALLEGHENY UNIVERSITY OF HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-03-01 至 1998-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2002455
• 关键词: CD antigens; atherosclerosis; cell adhesion molecules; cell line; cellular pathology; chimeric proteins; collagenase; cytogenetics; extracellular matrix; flow cytometry; gene expression; gene induction /repression; genetic regulation; integrins; low density lipoprotein receptor; macrophage; monocyte; pathologic process; thromboplastin; tissue /cell culture; transcription factor; transfection; western blottings

Macrophage participate or play a leading role in all stages of atherosclerosis. Therefore, identifying and regulating genes that code for macrophage functions in atherosclerotic lesions has the potential of preventing many aspects of this vascular disease. The present application proposes to adapt anti-DNA antibodies for the purpose of repressing genes that promote the atherosclerotic process and depend on the AP-1 transcription factor for expression. Preliminary data suggest that the AP-1 dependent transcription of the interstitial collagenase (MMP-1) gene is effectively repressed by intracellular fusion proteins consisting of a single-chain anti-DNA Fv fragment (scFv) and leucine zipper (Lz) domains derived from c-Jun or c-Fos. The goal of the present proposal is to extend these studies to other AP-l activated genes expressed by macrophage that are implicated in aspects of atherosclerosis. Expression vectors for anti-DNA scFv-LZ fusion proteins will be transfected into the human U937 and THP-1 monocytic cell lines as well as into purrried monocytes from peripheral blood. The expression of the genes for the following proteins will be investigated to establish the extent to which they are susceptible to AP-1 inhibition: 1. The beta2 integrin family adhesion molecule CD11c (p150,95) that is predominantly expressed on monocytes and macrophage and contributes to their adhesion to activated endothelial cells; 2. The matrix metalloprotease MMP-1 that is expressed in a cell tppe selective manner by monocytic cells and contributes to extracellular matrix (ECM) degradation and fibrous plaque rupture; 3. The scavenger receptor that becomes expressed as monocytes differentiate into foam cells and that mediates oxidized low density lipoprotein absorption; 4. Tissue factor, an enzyme expressed on the surface of foam cells in atherosclerotic lesions that acts as a cofactor for Factor VIIa and thus participates in the first reaction of the extiiinsic pathway of coagulation. There is preliminary evidence that each of these genes requires AP-1 for trnscription and therefore should be repressed by AP-1 inhibition. It is hoped that the proposed experiments will lay the foundation for the future testing of these treatments in animal models of atherosclerosis.

巨噬细胞在各个阶段都参与或起主导作用 动脉粥样硬化。因此，识别和调控编码的基因 巨噬细胞在动脉粥样硬化病变中的功能具有潜力 预防这种血管疾病的许多方面。现在的 申请建议采用抗 DNA 抗体，目的是 抑制促进动脉粥样硬化过程并依赖的基因 AP-1 转录因子的表达。初步数据 表明间质的 AP-1 依赖性转录 胶原酶（MMP-1）基因被细胞内有效抑制 由单链抗 DNA Fv 片段组成的融合蛋白 (scFv) 和亮氨酸拉链 (Lz) 结构域源自 c-Jun 或 c-Fos。这 本提案的目标是将这些研究扩展到其他 AP-l 巨噬细胞表达的激活基因与各方面有关 动脉粥样硬化。用于抗 DNA scFv-LZ 融合的表达载体 蛋白质将被转染到人类 U937 和 THP-1 单核细胞中 细胞系以及外周血中的咕噜单核细胞。 以下蛋白质的基因表达将是 进行调查以确定他们易受影响的程度 AP-1 抑制： 1. β2 整合素家族粘附分子 CD11c (p150,95) 主要表达于单核细胞和巨噬细胞，并有助于 它们与活化的内皮细胞的粘附； 2. 细胞tppe中表达的基质金属蛋白酶MMP-1 单核细胞的选择性方式并有助于细胞外 基质（ECM）降解和纤维斑块破裂； 3. 清道夫受体以单核细胞的形式表达 分化成泡沫细胞并介导氧化低密度 脂蛋白吸收； 4. 组织因子，一种在泡沫细胞表面表达的酶 动脉粥样硬化病变，作为因子 VIIa 和的辅助因子 因此参与了外源途径的第一个反应 凝固。 初步证据表明这些基因中的每一个都需要 AP-1 转录，因此应通过 AP-1 抑制来抑制。 希望所提出的实验能够为 未来在动物模型中测试这些治疗方法 动脉粥样硬化。