Protein therapeutics for the chorioretina

脉络膜视网膜的蛋白质疗法

• 批准号: 7594095
• 负责人: SOFIA P BECERRA
• 金额: $ 78.75万
• 依托单位: NATIONAL EYE INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7594095
• 关键词: Adverse effects; Angiogenesis Inhibitors; Angiostatins; Animal Model; Animals; Antibiotics; Binding; Cells; Choroid; Choroidal Neovascularization; Complex; Detection; Development; Devices; Diet; Disease; Disease regression; Docosahexaenoic Acids; Dose; Enzymes; Evaluation; Extracellular Matrix; Eye; Fluorescein; Fluoresceins; Freezing; Gelatinase A; Goals; Imaging Techniques; Injection of therapeutic agent; Laser injury; Lasers; Matrix Metalloproteinases; Measures; Microscopy; Molecular; Ovalbumin; Peptides; Phospholipases A; Prevention; Proteins; Rattus; Relative (related person); Research; Retina; Rodent; Route; Sclera; Serpins; Serum; Signal Transduction; Solutions; Specificity; Techniques; Temperature; Work; copolymer; day; design; enzyme activity; extracellular; in vivo; novel; pigment epithelium-derived factor; pigment epithelium-derived factor receptor; therapeutic protein

We completed the development of a novel imaging technique for the evaluation and quantification of laser-induced choroidal neovascularization (CNV) complexes in rodents. Using this technique we continued evaluating the efficacy of PEDF-derived peptides and other antiangiogenic factors on experimental CNV complex development. The route of delivery was a subconjunctival injection of protein solution. PEDF solutions were injected before and after laser injury or even after vessels had been formed. We found that PEDF, small peptides derived from the PEDF amino-end region, and angiostatin inhibited CNV development. The efficacy of PEDF subconjunctival injections on regression of fully developed vessels was observed. We continued the studies on the effects of extracellular matrix degrading enzymes on PEDF and PEDF-derived peptides. The MMP-2 enzyme activity on itself and on PEDF was pH sensitive. We also continued studying the effects of doxycyclin (bacteriostatic antibiotic with anti-metalloproteolytic effects) on CNV. Doxycyclin administered orally inhibited experimental CNV complex development. We determined the levels of matrix metalloproteinases type 2 and 9 and of PEDF in serum of doxy treated animals. We found that PEDF levels had increased with doxycyclin treatments. We evaluated potential matrixes for the ocular delivery of proteins. Subconjunctival injections of a temperature sensitive copolymer containing fluoresceinated ovalbumin (a serpin closely related to PEDF) were performed in rat eyes. Microscopy of frozen cross sections of eyes showed detection of fluorescein signal distributed in the sclera, choroid, RPE and retina even at 14 days post-injection. PEDF stimulates the phospholipase A activity of a PEDF receptor, and the release of DHA from RPE cells. The effects of dietary DHA in experimental CNV in vivo were examined. We measured the volumes of CNV complexes of rodents subjected to DHA deficient and adequate diets. CNV complex volumes in animals with the adequate diet were significantly decreased relative to those in animals with the DHA deficient diet.

我们完成了一种新型成像技术的开发，用于评估和定量啮齿动物中激光诱导的脉络膜新生血管形成（CNV）络合物。使用这种技术，我们继续评估PEDF衍生的肽和其他抗血管生成因子对实验CNV复合发育的疗效。递送途径是蛋白质溶液的结膜下注入。在激光损伤之前和之后，甚至在形成血管后，都注入PEDF溶液。我们发现，PEDF，源自PEDF氨基末端区域的小肽，Angiostatin抑制了CNV的发育。观察到PEDF亚结肠膜注射对完全发育血管回归的功效。 我们继续研究细胞外基质降解酶对PEDF和PEDF衍生肽的影响。 MMP-2酶的活性本身和PEDF对pH敏感。我们还继续研究强甲环蛋白（具有抗替代溶解作用的抑菌抗生素）对CNV的影响。多西环蛋白口服抑制实验CNV复合物的发育。我们确定了在Doxy治疗的动物的血清中，基质金属蛋白酶的水平以及PEDF的水平。我们发现，多西尔环蛋白治疗的PEDF水平增加了。 我们评估了蛋白质递送的潜在基质。 在大鼠眼中，对含有荧光卵蛋白的温度敏感共聚物（与PEDF密切相关的SERPIN）的温度敏感共聚物注入。冻结的眼睛横截面的显微镜显示，即使在注射后14天，也发现了巩膜，脉络膜，RPE和视网膜分布的荧光素信号。 PEDF刺激PEDF受体的磷脂酶A活性，并从RPE细胞中释放DHA。 检查了饮食DHA在体内实验性CNV中的影响。我们测量了受DHA不足和足够饮食的啮齿动物CNV复合物的体积。 相对于DHA缺乏饮食的动物，足够饮食的动物中的CNV复合体积显着减少。