BETA 1 INTEGRIN ANTAGONISM & RETINAL NEOVASCULARIZATION

Beta 1 整合素拮抗作用

• 批准号: 6350899
• 负责人: DONALD R SENGER
• 金额: $ 17.51万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6350899
• 关键词: angiogenesis; cell adhesion; diabetic retinopathy; immunoprecipitation; in situ hybridization; inhibitor /antagonist; integrins; laboratory mouse; monoclonal antibody; northern blottings; protein structure function; receptor expression; retina; tissue /cell culture; vascular endothelial growth factors; vascular endothelium

DESCRIPTION: (Applicant's Description) The broad long term objective of this collaborative research plan is to identify new strategies for controlling the retinal neovascularization (NV) that leads to blindness in patients with diabetic retinopathy, age-related macular degeneration, and retinopathy of prematurity. Vascular endothelial growth factor (VEGF), a potent hypoxia-inducible angiogenic cytokine, has been implicated causally in retinal NV suggesting that VEGF and the mechanisms by which VEGF promotes angiogenesis are important targets for therapy. VEGF markedly induces expression of the alpha-1-beta-1, alpha-2-beta-1, and alpha- 5-beta-3 cell surface integrins in dermal microvascular endothelial cells (ECs), and antagonism of alpha-1-beta-1, alpha-2-beta-1, and alpha-5 integrins inhibits VEGF-driven angiogenesis in skin. In addition, targeting of alpha-5 integrins has shown promise for inhibition of retinal NV. However, the significance of alpha-1-beta-1, alpha-2-beta-1, and other beta-1 integrins for retinal NV has not been investigated. Therefore, the proposed experimental plan will: (1) investigate VEGF regulation of integrin expression by retinal ECs in vitro with particular emphasis on beta-1 integrins, and determine if the same integrin mRNAs induced by VEGF in vitro are induced similarly during retinal NV in vivo; and (2) investigate inhibition of retinal NV in vivo by monoclonal antibodies (Abs) that block alpha-1-beta-1 and alpha-2-beta-1 integrins and monoclonal Abs that block other beta-1 integrins induced in retinal ECs by VEGF. Northern analyses will test whether and to what extent VEGF, similar to its action towards dermal microvascular ECs, induces isolated bovine retinal ECs to express mRNAs encoding the alpha-1 and alpha-2 integrin subunits. Also, cell surface labeling and immunoprecipitation will be employed to determine if VEGF induces retinal ECs to express the alpha-1-beta-1 and alpha- 2-beta-1 heterodimers at the cell surface. Similarly, VEGF regulation of other beta-1 integrins in retinal ECs will also be investigated. In situ hybridization analyses of a well-defined mouse model of proliferative refinopathy will determine whether integrin subunit mRNAs induced by VEGF in vitro are similarly induced in ECs during retinal NV in vivo. Finally, specific monoclonal Abs which independently block alpha-1-beta-1, alpha-1- beta-1, and other beta-1 integrins induced by VEGF in retinal ECs will be tested for inhibition of retinal NV in vivo. Thus far, the importance of beta-1 integrins for proliferative retinopathies has not been explored. The proposed experiments will fill major gaps in knowledge by providing crucial information on the significance of beta-1 integrins for retinal NV. Moreover, it is anticipated that findings here will suggest new strategies for therapeutic intervention.

描述：（申请人的描述） 这项合作研究计划的广泛长期目标是 确定控制视网膜新生血管（NV）的新策略 导致糖尿病视网膜病变患者失明， 黄斑变性和早产儿视网膜病。 血管内皮 生长因子（VEGF）是一种有效的缺氧诱导的血管生成细胞因子， 与视网膜NV有因果关系，表明VEGF及其机制 VEGF促进血管生成是治疗的重要靶点。 VEGF 显著诱导α-1-β-1、α-2-β-1和α- 真皮微血管内皮细胞5-β-3细胞表面整合素 (ECs)和α-1-β-1、α-2-β-1和α-5整联蛋白的拮抗作用 抑制皮肤中VEGF驱动的血管生成。 此外，针对阿尔法-5 整联蛋白已显示出抑制视网膜NV的前景。 但 α-1-β-1、α-2-β-1和其它β-1整联蛋白对于 视网膜NV尚未被研究。 因此，建议的实验 计划：（1）研究VEGF对视网膜整合素表达的调节， 特别强调β-1整合素的体外EC，并确定 在体外由VEGF诱导的相同整合素mRNA在 体内视网膜NV;和（2）研究体内视网膜NV的抑制作用 阻断α-1-β-1和α-2-β-1的单克隆抗体（Abs） 整合素和单克隆抗体，其阻断在细胞中诱导的其它β-1整合素， 视网膜内皮细胞VEGF。 北方分析将测试是否以及在多大程度上VEGF，类似于 其对真皮微血管内皮细胞的作用，诱导分离的牛视网膜 EC表达编码α-1和α-2整联蛋白亚基的mRNA。 此外，细胞表面标记和免疫沉淀将被用于 确定VEGF是否诱导视网膜EC表达α-1-β-1和α-1-β-1 2-β-1异源二聚体。 类似地，VEGF调节 还将研究视网膜EC中的其它β-1整联蛋白。 原位 明确定义的增殖性小鼠模型的杂交分析 胶质瘤病将决定VEGF诱导的整合素亚基mRNA是否在胶质瘤中起作用。 在体内视网膜NV过程中，在EC中类似地诱导体外细胞。 最后， 特异性单克隆抗体，其独立地阻断α-1-β-1，α-1- 在视网膜EC中由VEGF诱导的β-1和其他β-1整联蛋白将被 测试体内视网膜NV的抑制。 到目前为止， β-1整联蛋白对增殖性视网膜病的作用还没有研究。 的 拟议的实验将填补知识的主要空白，提供关键的 关于β-1整合素对视网膜NV的意义的信息。 此外，委员会认为， 预计，这里的发现将提出新的战略， 治疗干预