Dietary control of angiogenesis in retinopathy model; basis for clinical trials

描述(由申请人提供)：这项提案描述了一种创新的营养/药理学策略，以预防小鼠疾病模型中的增殖性视网膜病变，并期望在以后的糖尿病视网膜病变(DR)患者的临床试验中使用。DR有炎症和血管生成两种成分。我们将单独和交互评估？-3长链多不饱和脂肪酸(LCPUFAs)二十二碳六烯酸和二十碳五烯酸以及选择性环氧合酶(COX)-2抑制剂塞来昔布的疗效，Celecoxib是一种选择性环氧合酶(COX)-2抑制剂，旨在针对与增殖性DR？-3LCPUFAs有关的炎症和血管生成因子。？-3LCPUFAs集中在视网膜中，这种状态可以通过饮食摄入量改变，并且依赖于从所有西方饮食中摄入的食物，因此这些脂类是预防Dr.Dr.的干预措施的合理选择。初步结果显示，COX-2抑制剂可以预防缺血诱导的视网膜新生血管，并且-2抑制与？-3LCPUFAs具有协同作用。如果塞来昔布被证明会导致心脏病，我们将改用阿司匹林(COX 1，2抑制药)。为了确定抑制机制和确定补充干预措施，将使用一种新技术来敏感地测量治疗过程中特定脂质、炎症和血管生成途径的mRNAs的绝对拷贝数的微小变化。该小组将包括(但不限于)COX1，2和LOX，血管内皮生长因子和受体1，2，细胞黏附分子(ICAM-1，CD18)，基质金属蛋白酶2，9，细胞因子(TNFa，TGF？)，核激素受体(NF？B)和一氧化氮合酶。为了评估干预措施和可能的DR新的替代标记物，我们将评估摄入平衡、等热量饮食的小鼠视网膜和血液中？-3LCPUFAs、花生四烯酸和二十碳二烯(血栓烷、前列腺素、白三烯)的水平。在这项翻译工作中，已经成立了一个机构间和学科内的研究小组，成员包括以下领域的专家：分子和细胞生物学与血管生成(L·史密斯)、临床研究(E Chew、J-P SanGiovanni，NEI)、营养与脂质生物化学(N Salem、NIAAA、C Serhan、哈佛大学)。如果成功，这项工作可能会对预防增生性DR产生巨大影响，因为干预措施安全、廉价，而且易于实施。初步结果表明，COX-2抑制剂、β-3 LCPUFAs及其联合应用可抑制视网膜病变。这些研究的创新之处在于：1.他们首次检测了在使用或不使用COX-2抑制剂的情况下，LCPUFA对DR的影响。2.一种新的技术，即在疾病和干预期间发现mRNA的绝对拷贝数，允许比较干预措施，以评估补充治疗。3.血脂分析可能产生DR和Risk的机制和新的替代标记物。

DESCRIPTION (provided by applicant): This proposal describes an innovative nutritional/pharmacological strategy to prevent proliferative retinopathy in a mouse model of disease with the expectation of a later clinical trial of patients with diabetic retinopathy (DR). DR has inflammatory and angiogenic components. We will evaluate alone and interactively the efficacy of the ?-3 long-chain polyunsaturated fatty acids (LCPUFAs) docosahexaenoic acid and eicosapentaenoic acid and Celecoxib, a selective cyclooxygenase (COX)-2 inhibitor, designed to target inflammatory and angiogenic factors implicated in the pathogenesis of proliferative DR. ?-3 LCPUFAs are concentrated in the retina and the status is modifiable by and dependent on dietary intake from foods that are not consumed in all Western diets, so these lipids are reasonable choices for interventions to prevent DR. Preliminary results show COX-2 inhibitors prevent ischemia-induced retinal neovascularization and that COX-2 inhibition is synergistic with ?-3 LCPUFAs. Should Celecoxib prove to cause cardiac disease we will switch to aspirin (COX 1,2 inhibition). To determine mechanism of inhibition and to determine complementary interventions, a new technique will be used to sensitively measure small changes with treatment in absolute copy number of mRNAs of specific lipid, inflammatory and angiogenic pathways involved in disease. The panel will include (but is not limited to) COX1,2, and LOX, vascular endothelial growth factor and receptors 1,2, cell adhesion molecules (ICAM-1, CD18), matrix metalloproteinases 2,9, cytokines (TNFa, TGF?), nuclear hormone receptor (NF?B), and nitric oxide synthetase. To assess intervention and possible new surrogate markers of DR we will assess retinal and blood levels of ?-3 LCPUFAs, arachidonic acid and eicosanoids (thromboxanes, prostaglandins, leukotrienes) in mice consuming balanced, isocaloric diets with, or without ?-3 LCPUFAs. For this translational work an inter-institutional and intra-disciplinary research group has been established with experts in the fields of molecular and cellular biology and angiogenesis (L Smith, (PI), D Carper, J-Y Tsai, NEI), clinical research (E Chew, J-P SanGiovanni, NEI), nutrition and lipid biochemistry (N Salem, NIAAA, C Serhan, Harvard). If successful, this work could have a great impact on preventing proliferative DR, as the interventions are safe, inexpensive, and can be easily implemented. Preliminary results show that COX-2 inhibitors, ?-3 LCPUFAs and combination inhibit retinopathy. These studies are innovative in that: 1. they are the first to examine the effect of LCPUFAs with or without COX-2 inhibitors on DR. 2. a new technique of finding absolute copy numbers of mRNA during disease and intervention allows comparison of interventions to evaluate complementary treatments. 3. lipid analysis may yield mechanism as well as new surrogate markers of DR and risk.