Fragments of TrpRS to Treat Neovascular Eye Diseases

描述（由申请人提供）：绝大多数导致灾难性视力丧失的疾病是由于异常血管生成。病理性视网膜或脉络膜新生血管分别导致糖尿病视网膜病变（DR）和年龄相关性黄斑变性（ARMD）的视力丧失。虽然抑制异常血管生成不一定能治愈潜在的疾病，但它可以通过预防与新血管形成相关的并发症（如出血和水肿）来保护视力。我们一直在研究的抗血管生成活性的片段的右旋糖氨酰-tRNA合成酶（TrpRS）。在正常人细胞中，TrpRS以全长形式和截短形式（mini-TrpRS）存在，其中氨基末端结构域由于前体mRNA的选择性剪接而缺失。后一种形式优先在暴露于干扰素-w的细胞中合成。mini-TrpRS的进一步截短产生42 kD形式（T2），其是迄今为止评价的TrpRS的血管抑制形式中最有效的。 在本申请中，我们提出进一步表征TrpRS片段的抗血管生成活性，并鉴定用于新生血管性眼病临床试验的候选药物和递送系统。具体而言，我们将：（1）检查TrpRS片段在正常和异常眼血管生成的调节中的生理作用;（2）鉴定和表征这些片段结合的视网膜受体;（3）表征具有抗血管生成活性的TrpRS片段的结构方面，并使用该信息来模拟具有类似活性的小分子拮抗剂;（4）开发基于病毒、细胞和靶向脂质体的载体，用于递送T2以在多种动物模型中抑制眼部新生血管形成;和（5）开始对这些载体进行药代动力学和毒理学研究，重组蛋白和/或小分子治疗剂作为人类临床试验的第一步，用于治疗新生血管性眼病，如新生血管性ARMD、增殖性DR和红变性青光眼。

DESCRIPTION (provided by applicant): The vast majority of diseases that cause catastrophic loss of vision do so as a result of abnormal angiogenesis. Pathological retinal or choroidal neovascularization lead to visual loss in diabetic retinopathy (DR) and age related macular degeneration (ARMD), respectively. While inhibition of abnormal angiogenesis would not necessarily cure the underlying diseases, it would preserve vision by preventing complications associated with neovascularization such as hemorrhage and edema. We have been studying the anti-angiogenic activity of fragments of tryptophanyl-tRNA synthetase (TrpRS). In normal human cells TrpRS exists as both the full length form and a truncated form (mini-TrpRS) in which an amino-terminal domain is deleted due to alternative splicing of the pre-mRNA. This latter form is preferentially synthesized in cells exposed to interferon-w. Further truncation of mini-TrpRS results in a 42 kD form (T2) that is the most potent of the angiostatic forms of TrpRS evaluated to date. In this application we propose to further characterize the anti-angiogenic activity of TrpRS fragments and identify a candidate drug and delivery system for use in clinical trials of neovascular eye diseases. Specifically, we will: (1) examine the physiological role of TrpRS fragments in the regulation of normal and abnormal ocular angiogenesis; (2) identify and characterize the retinal receptor to which these fragments bind; (3) characterize the structural aspects of TrpRS fragments with anti-angiogenic activity and use this information to model small molecular antagonists with similar activity; (4) develop viral-, cell- and targeted liposome-based vectors for the delivery of T2 to inhibit ocular neovascularization in a variety of animal models; and (5) begin pharmacokinetic and toxicology studies on these vector, recombinant protein and/or small molecule therapeutics as a first step towards human clinical trials for the treatment of neovascular eye diseases such as neovascular ARMD, proliferative DR and rubeotic glaucoma.