AG13764 and AG13711 Reverses VEGF-Induced Choroidal Neovascularization in Rat Eye

AG13764 和 AG13711 逆转 VEGF 诱导的大鼠眼脉络膜新生血管形成

基本信息

批准号：
7734628
负责人：
SHELDON S MILLER
金额：
$ 5.31万
依托单位：
NATIONAL EYE INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

项目摘要

Purpose: Age-related macular degeneration (AMD) is the major cause of blindness for people over 60. In the wet form of AMD compounds targeting growth factor signaling pathways such as VEGF have been a major focus for therapeutic interventions. In a previously developed rat model of CNV, we utilized two receptor tyrosine kinase inhibitors (RTKi) to block VEGFR-1, VEGFR-2 and PDGFR signaling following the establishment of CNV. Methods: AAV-VEGF165 was injected into the subretinal space of rats at postnatal days 15-17. Six weeks later, a suspension of RTK inhibitors, AG013764 or AG013711, was injected intraperitoneally (IP, twice daily) or intravitreally (every five days) over a two week period. FITC-dextran whole-mounts of RPE-choroid-sclera were prepared after the animals were sacrificed. CNV area was quantified using Neurolucida to measure the hyperfluorescence on FITC-dextran whole-mounts. Histology and immunohistochemistry were performed as described previously. Results: VEGF expression in control and treated eyes was confirmed by immunohistochemistry and histological sections indicated recovery of retinal morphology and CNV reduction in treated eyes. In the animals injected by IP with AG013764 or AG013711 the mean CNV level was reduced by 25 to 33% compared to control, but this effect did not achieve statistical significance. Intravitreal injections of AG013764 or AG013711 reduced the level of CNV by approximately 60% compared to control (p< 0.005 or p< 0.05, respectively). Conclusions: These data show that two RTK inhibitors, AG013764 or AG013711, delivered intravitreally, significantly reduce blood vessel proliferation in this AAV-VEGF165 model of CNV.

目的：与年龄相关的黄斑变性（AMD）是60岁以上患者失明的主要原因。在AMD化合物的潮湿形式中，靶向生长因子信号传导途径（例如VEGF）一直是治疗干预措施的主要重点。在先前开发的CNV大鼠模型中，我们利用了两个受体酪氨酸激酶抑制剂（RTKI）来阻止CNV建立后VEGFR-1，VEGFR-2和PDGFR信号传导。方法：在产后15-17天，将AAV-VEGF165注入大鼠的视网膜下空间。六个星期后，在两周内对RTK抑制剂，AG013764或AG013711的悬浮液被腹膜内（每天两次）或玻璃体内（每五天）注射。在牺牲动物后，制备了FITC-DEXTRAN的RPE-螺状螺旋体的整体。使用Neurolucida对CNV区域进行定量，以测量FITC-DEXTRAN全座上的超荧光。如前所述进行了组织学和免疫组织化学。结果：通过免疫组织化学证实了对照和治疗的眼睛中的VEGF表达，组织学切片表明，治疗的眼睛中的视网膜形态和CNV减少了。与对照相比，在IP注入AG013764或AG013711的动物中，平均CNV水平降低了25％至33％，但这种效应没有达到统计学意义。与对照相比，玻璃体内注射AG013764或AG013711的CNV水平降低了约60％（分别为P <0.005或P <0.05）。结论：这些数据表明，在这种AAV-VEGF165的CNV模型中，两种RTK抑制剂AG013764或AG013711显着降低了血管增殖。