Eicosanoid Gene Therapy

项目描述（申请人提供）：本项目为原发性开角型青光眼（POAG）提供慢病毒载体基因治疗。在之前的资助周期中，我们建立了一个强大的载体平台和实验能力，用于将基因传递到房水动力学中涉及的前房结构。我们发现慢病毒载体可以有效且无毒地转导小梁网，并实现与POAG的慢性性相称的持续表达。建立了家猫和食蟹猴两种大型动物模型。本研究探讨了类二十烷的基因治疗潜力，类二十烷是一种强有力的脂质衍生信号分子，包括前列腺素、血栓烷和白三烯。我们假设，由于PGF21激动剂已被证明具有降低眼压（IOP）的活性，我们可以通过基因治疗来操纵眼睛中的前列腺素生理，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，最初的重点是环氧化酶-2 （COX-2）和PGF21途径，因为它在IOP生理中已知的作用，并成功地作为一个药理学靶点（latanoprost和相关药物）。我们获得了合成的、序列优化的cdna，这些cdna对于克服极端的mRNA依赖性至关重要。这种基因治疗导致猫持续的、无毒的IOP降低。我们将开发和使用慢病毒载体在组织培养试验、灌注的人类供体眼睛和两种动物模型中，以建立一种实用的、完全翻译的降低眼压的基因疗法。将系统地研究先进的慢病毒载体表达变量和替代PG途径，以了解类二十烷醇对前房生理的影响，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基因治疗原发性开角型青光眼（POAG）。我们将确定最佳的前列腺素生物合成和持续IOP降低的反应途径，我们将获得关于前列腺素在眼睛中的作用的新知识。

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.