Eicosanoid Gene Therapy

描述（由申请人提供）：本项目将为原发性开角型青光眼（POAG）提供基于慢病毒载体的基因治疗。在上一个资助周期中，我们建立了一个强大的载体平台和实验能力，用于将基因递送到涉及房水动力学的前房结构。我们发现，慢病毒载体可以有效地和非毒性的小梁网，并实现持久的表达相称的慢性POAG。在家猫和食蟹猴中建立了两种大型动物模型。在这里，我们探索了类花生酸的基因治疗潜力，类花生酸是一种有效的脂质衍生信号分子，包括类花生素、血栓烷和白三烯。我们假设，由于PGF 21激动剂已被证明具有降低眼内压（IOP）的活性，我们可以通过基因治疗来操纵眼内的前列腺素生理学，以实现持续的IOP降低。在初步研究中，我们在体外和体内设计了一个完整的前列腺素生物合成和反应途径，最初关注环氧合酶-2（考克斯-2）和PGF 21途径，因为其在IOP生理学中的已知作用及其作为药理学靶点（拉坦前列素和相关药物）的成功开发。我们得到了合成的，序列优化的cDNA，这是克服极端mRNA的责任。这种基因治疗的结果在持续的，无毒的眼压降低猫。我们将开发和使用慢病毒载体在组织培养试验，灌注人供体的眼睛，和两个动物模型，以建立一个实用的，完全翻译的眼压降低基因治疗。将系统研究先进的慢病毒载体表达变量和替代PG途径，以了解类花生酸对前房生理学的影响，并最大限度地发挥基因治疗的潜力。公共卫生相关性。 该项目将开发基于慢病毒载体的原发性开角型青光眼（POAG）基因治疗。我们将确定持续降低IOP的最佳前列腺素生物合成和反应途径，我们将获得有关眼内前列腺素作用的新知识。

DESCRIPTION (provided by applicant): This project will provide lentiviral vector-based gene therapy for primary open angle glaucoma (POAG). In the previous funding cycle we established a robust vector platform and experimental capacity for gene delivery to anterior chamber structures involved in aqueous humor dynamics. We showed that lentiviral vectors could effectively and non-toxically transduce the trabecular meshwork and achieve sustained expression commensurate to the chronicity of POAG. Two large animal models were established in the domestic cat and Cynomolgus monkey. Here we have explored the gene therapy potential of eicosanoids, the potent lipid- derived signaling molecules that include the prostaglandins, thromboxanes and leukotrienes. We hypothesize that since PGF21 agonists have proven intraocular pressure (IOP)-lowering activity, we can manipulate prostaglandin physiology in the eye via gene therapy to achieve sustained IOP reduction. In preliminary studies, we engineered a complete prostaglandin biosynthesis and response pathway in vitro and in vivo, with initial focus on cyclooxygenase-2 (COX-2) and on the PGF21 pathway because of its known role in IOP physiology and its successful exploitation as a pharmacologic target (latanoprost and related drugs). We derived synthetic, sequence optimized cDNAs that were essential to overcoming extreme mRNA liability. This gene therapy results in sustained, non-toxic IOP reduction in the cat. We will develop and use lentiviral vectors in tissue culture assays, perfused human donor eyes, and the two animal models to establish a practical, fully translational IOP-reduction gene therapy. Advanced lentiviral vector expression variables and alternative PG pathways will be systematically studied to learn about the effects of eicosanoids on anterior chamber physiology and maximize gene therapy potential. PUBLIC HEALTH RELEVANCE. The project will develop lentiviral vector-based gene therapy for primary open angle glaucoma (POAG). We will determine optimal prostaglandin biosynthesis and response pathways for sustained IOP reduction and we will gain new knowledge about the roles of prostaglandins in the eye.