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

该提案描述了一种创新的营养/药理学策略，以预防增殖性视网膜病变 在小鼠疾病模型中进行，并期望在以后的糖尿病视网膜病变（DR）患者的临床试验中进行。 DR具有炎症和血管生成成分。我们将单独和交互地评估co-3的功效。 长链多不饱和脂肪酸（LCPUFA）、二十二碳六烯酸和二十碳五烯酸，以及 塞来昔布是一种选择性环氧合酶（考克斯）-2抑制剂，旨在靶向炎症和血管生成因子 参与增殖性DR的发病机制。co-3LCPUFA集中在视网膜中， 可以通过和依赖于饮食摄入的食物来改变，这些食物在所有西方饮食中都没有被消费，所以这些 脂质是预防DR干预的合理选择。初步结果显示，考克斯-2抑制剂可预防 缺血诱导视网膜新生血管形成，且考克斯-2抑制与co-3 LCPUFA具有协同作用。应该 塞来昔布被证明会导致心脏疾病，我们将改用阿司匹林（考克斯1，2抑制剂）。 为了确定补充干预措施，将使用一种新技术来灵敏地测量 特定脂质、炎症和血管生成的mRNA的绝对拷贝数随治疗的微小变化 参与疾病的途径。该组将包括（但不限于）COX 1、2和LOX，血管 内皮生长因子及其受体1、2、细胞粘附分子（ICAM-1、CD 18）、基质金属蛋白酶 2，9，细胞因子（TNF α，TGF β），核激素受体（NF κ B）和一氧化氮合成酶。评估 干预和可能的新DR替代标志物我们将评估co-3 LCPUFA的视网膜和血液水平， 花生四烯酸和类二十烷酸（血栓烷，白藜芦醇，白三烯）在小鼠消耗平衡， 含或不含co-3LCPUFA的等热量饮食。对于这项翻译工作的机构间和内部- 学科研究小组已成立，专家在分子和细胞生物学领域， 血管生成（L Smith，（PI），D Carper，J-Y Tsai，NEI），临床研究（E Chew，J-P SanGiovanni，NEI）， 营养和脂质生物化学（N Salem，NIAAA，C Serhan，哈佛）， 如果成功，这项工作可能对预防增殖性DR产生巨大影响，因为干预措施是安全的， 价格便宜，并且可以容易地实现。初步结果表明，考克斯-2抑制剂，co-3 LCPUFA和 组合抑制视网膜病。这些研究的创新之处在于：1 .首次探讨了 含或不含考克斯-2抑制剂的LCPUFAs对DR的影响2.一种发现DR绝对拷贝数的新技术 疾病和干预期间的mRNA允许比较干预措施以评估补充治疗。 3.脂质分析可能产生机制以及新的DR和风险的替代标志物。

This proposal describes an innovative nutritional/pharmacological strategy to prevent proliferative retinopathy in a mouse model of disease with the expectation of a later clinicaltrial of patients with diabetic retinopathy(DR). DR has inflammatory and angiogenic components. We will evaluate alone and interactively the efficacy of the co-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. co-3LCPUFAs 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 inhibitionis synergistic with co-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 (butis 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.TGFp), nuclear hormone receptor (NFicB), and nitric oxide synthetase. To assess intervention and possible new surrogate markers of DR we will assess retinal and blood levels of co-3 LCPUFAs, arachidonic acid and eicosanoids (thromboxanes, prostaglandins, leukotrienes) in mice consuming balanced, isocaloric diets with, or without co-3LCPUFAs. 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, co-3LCPUFAs 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.