To Treat Some Ocular Diseases with Id1/Id3 Inhibitors

用 Id1/Id3 抑制剂治疗一些眼部疾病

• 批准号: 6834194
• 负责人: CAROL L MESCHTER
• 金额: $ 10.25万
• 依托单位: COMPARATIVE BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2006-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6834194
• 关键词: RNA interference; angiogenesis; angiogenesis inhibitors; antisense nucleic acid; diabetic retinopathy; drug delivery systems; drug design /synthesis /production; eye disorder chemotherapy; gene induction /repression; hyperoxia; laboratory mouse; macular degeneration; nonhuman therapy evaluation; oligonucleotides; small interfering RNA

The Id1 and Id3 genes and proteins are critical components of the angiogenic process and participate in a wide range of processes involved in cellular differentiation and proliferation. It is now known that Id proteins help regulate the release and recruitment of bone marrow derived and circulating endothelial precursor cells which help form the new blood vessels of tumors. Recently this same observation has been extended to neovascularization in the eye. For this reason, we propose that either preventing expression of Id1/Id3 genes or inhibiting the functions of the Id1 and Id3 proteins in the eye could be an effective way to prevent or treat ocular neovascularization. Inhibiting pathologic ocular angiogenesis is of critical importance because aberrant blood vessel formation leads to severe visual loss in age-related macular degeneration (ARMD), diabetic retinopathy (DR) and retinopathy of prematurity (ROP). We, and one other group, have successfully used an antisense molecule to Id1 to block angiogenesis in an animal model unrelated to the eye. Such an agent might be useful if administered intravitreally to the eye. In addition to its therapeutic potential, an antisense molecule would also find significant use as a tool for proof-of-concept experiments in animal models of human ocular disease. Thus, the over-all goal of the proposed research is to confirm that inhibition of Id1/Id3 gene expression is a useful approach for treating ocular diseases characterized by pathologic neovascularization. To accomplish these goals, we propose the following three specific aims: (1) Examine the role of Id1/Id3 in retinal angiogenesis by quantifying retinal neovascularization in a mouse model of ischemia-induced retinal neovascularization in mice deficient in Id1 and Id3 and by using an intravitreally delivered anti-DNA antisense oligonucleotide with known activity for blocking Idl expression and angiogenesis to duplicate the anti-angiogenic phenotype of the Id1/Id3 knockout mouse, (2) Use in vitro/in vivo Matrigel studies and the mouse hyperoxia model to test the activity of siRNA oligonucleotides and one additional antisense molecule, and (3) Optimize one or more DNA-based or siRNA acting oligonucleotides that demonstrate promising activity by evaluating various vehicles to improve the duration and effect of the agent and establishing a dose-response relationship for at least one of the active oligonucleotides. Accomplishing these aims will demonstrate the feasibility of using specific oligonucleotides to inhibit ocular angiogenesis by blocking Id1 expression. Phase II activities will expand the approach to include blocking Id3 expression, exploring the possibility of blocking both Id1 and Id3 expression and further optimization of the oligonucleotides and possibly oligonucleotide combinations in terms of activity, pharmacokinetics and bioavailability. In addition, candidate oligonucleotides will be evaluated in one or more additional animal models of human ocular disease such as the argon laser photocoagulation model in mice.

Id 1和Id 3基因和蛋白是血管生成过程的关键组分，并参与细胞分化和增殖所涉及的广泛过程。现在已知Id蛋白有助于调节骨髓来源的和循环的内皮前体细胞的释放和募集，所述内皮前体细胞有助于形成肿瘤的新血管。最近，同样的观察结果已经扩展到眼睛中的新血管形成。因此，我们提出，阻止Id 1/Id 3基因的表达或抑制眼内Id 1和Id 3蛋白的功能可能是预防或治疗眼内疾病的有效方法。 眼部新生血管抑制病理性眼部血管生成至关重要，因为异常血管形成导致年龄相关性黄斑变性（ARMD）、糖尿病视网膜病变（DR）和早产儿视网膜病变（ROP）中的严重视力丧失。我们和另一个小组已经成功地使用Id 1的反义分子来阻断与眼睛无关的动物模型中的血管生成。如果玻璃体内施用至眼睛，这样的药剂可能是有用的。除了其治疗潜力之外，反义分子还将作为在人类眼部疾病的动物模型中进行概念验证实验的工具而发现重要用途。因此，所提出的研究的总体目标是证实Id 1/Id 3的抑制 基因表达是治疗以病理性眼病为特征的眼病的有用方法， 新生血管形成为实现这些目标，我们提出以下三个具体目标：（1）通过在Id 1和Id 3缺陷的小鼠中定量缺血诱导的视网膜新生血管形成的小鼠模型中的视网膜新生血管形成，并通过使用具有已知的阻断Id 1表达和血管生成的活性的玻璃体内递送的抗DNA反义寡核苷酸来复制抗DNA反义寡核苷酸，来检查Id 1/Id 3在视网膜血管生成中的作用。Id 1/Id 3敲除小鼠的血管生成表型，（2）使用体外/体内Matrigel研究和小鼠高氧模型来测试siRNA寡核苷酸和一种另外的反义分子的活性，和（3）优化一个或多个DNA-通过评估各种载体来改善试剂的持续时间和效果，和建立至少一种活性寡核苷酸的剂量-反应关系。 实现这些目标将证明使用特异性寡核苷酸通过阻断Id 1表达来抑制眼部血管生成的可行性。II期活动将扩展该方法，包括阻断Id 3表达，探索阻断Id 1和Id 3表达的可能性，并进一步优化寡核苷酸和可能的寡核苷酸组合的活性，药代动力学和生物利用度。此外，将在一种或多种另外的人类眼部疾病的动物模型中评估候选寡核苷酸，例如小鼠中的氩激光光凝模型。