Oral therapy for diabetic retinopathy using ACE2/Ang1-7 bioencapsulated in plant cells

使用生物封装在植物细胞中的 ACE2/Ang1-7 口服治疗糖尿病视网膜病变

• 批准号: 9128010
• 负责人: HENRY DANIELL
• 金额: $ 40.7万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9128010
• 关键词: Acids; Alzheimer' s Disease; Amyloid; Angiotensin II; Animal Model; Attenuated; Bioreactors; Blood; Blood Circulation; Blood Vessels; Brain; Cardiovascular system; Cell Wall; Cells; Chloroplasts; Chorea; Clinical Trials; Complications of Diabetes Mellitus; Data; Development; Diabetes Mellitus; Diabetic Retinopathy; Edible Plants; Encapsulated; Enzymes; Equilibrium; Foundations; Functional disorder; Gene Delivery; Gene Expression; Generations; Genes; Genome; Glucose; Goals; Health; Human; Hyperactive behavior; Inflammation; Inflammatory; Insulin Resistance; Kidney Diseases; Lettuce - dietary; Lipids; Maintenance; Mediating; Microbe; Mus; Myelin Basic Proteins; Neuraxis; Oral; Organ; Oxidative Stress; Pathologic; Pathology; Pathway interactions; Patients; Peptidyl-Dipeptidase A; Physiology; Plant Leaves; Plants; Prevention; Production; Proteins; Renin-Angiotensin System; Retina; Retinal; Retinal Diseases; Retinal Ganglion Cells; Risk Factors; Role; Signal Pathway; Stomach; Testing; Therapeutic; Tissues; Toxin; Transgenic Organisms; Treatment Efficacy; Vascular Diseases; base; cell type; clinical application; clinical investigation; commensal microbes; cost; diabetic; diabetic patient; endothelial dysfunction; equilibration disorder; glucose metabolism; improved; in vivo; innovation; lipid metabolism; member; neurovascular; novel; novel strategies; oxidative damage; prevent; receptor; receptor mediated endocytosis; relating to nervous system; therapeutic protein; translational study; uptake; vascular inflammation

 DESCRIPTION: Diabetic retinopathy (DR) is the most common diabetic complication developing in both type1 and type2 diabetic patients and insofar is incurable. Hyperactivity of the renin angiotensin system (RAS), resulting in elevated concentrations of Angiotensin II (Ang II), is central to pathways leading to increased vascular inflammation, oxidative stress and endothelial dysfunction in diabetes and its associated complications including DR. The discovery of ACE2 established the existence of a protective axis of the RAS as a key regulator counteracting the deleterious effects of Ang II. We have recently demonstrated that increased expression of ACE2 and Ang1-7, the two key members of the protective axis, via AAV-mediated retinal gene delivery reduced diabetes-induced retinal pathology, validating that enhancing the ACE2/Ang1-7/MAS axis is a promising approach for treating patients with DR. In addition, enhancing Ang1-7 and ACE2 also improves lipid and glucose metabolism, ameliorates insulin resistance and confers protection against a variety of pathological conditions including diabetes-induced nephropathy and cardiovascular dysfunction. Thus an ideal strategy would be to enhance ACE2/Ang1-7 both systemically and locally at target tissues. This proposal takes advantage of an innovative concept based on the use of plant chloroplasts as highly efficient bioreactors for production and oral delivery of therapeutic proteins bio encapsulated in plant cells. The therapeutic protein is fused to chorea toxin subunit B, which not only facilitate the transmucosal transport into circulation, but also enhance tissue cell uptake by GM1 receptor mediated endocytosis. We have recently shown that orally delivered therapeutic proteins are widely distributed in organs including brain and retina crossing blood brain/retina barriers; oral delivery of bio encapsulated myelin basic protein ameliorated amyloid burden in advanced human and mouse Alzheimer's disease brains and retinas, prevented loss of retinal ganglion cells. Thus this novel concept provides a more efficient, low cost production and patient-friendly delivery of therapeutic proteins, particularly to the central nervous system, which has long been a major challenge. The goals of this proposal are to evaluate the efficacy of oral delivery of Ang1- 7 and ACE2 bio encapsulated in plant cells in prevention and treatment of DR in suitable animal models and to generate and characterize transplastomic ACE2 and Ang1-7 expression in edible lettuce in order to advance bio encapsulated protein delivery approach towards clinical investigations. Result from these proposed studies will provide proof-of-concept on feasibility and efficacy in animal models of DR using oral delivery of plant chloroplast- based therapeutic proteins, and set the foundation for initiating clinical trials by oral delivery of bio encapsulate ACE2 and Ang1-7 for prevention and treatment of DR.

 描述：糖尿病视网膜病变 (DR) 是 1 型和 2 型糖尿病患者中最常见的糖尿病并发症，并且目前无法治愈。肾素血管紧张素系统 (RAS) 过度活跃，导致血管紧张素 II (Ang II) 浓度升高，是导致糖尿病及其相关并发症（包括 DR）血管炎症、氧化应激和内皮功能障碍增加的途径的核心。 ACE2 的发现证实了 RAS 保护轴的存在，作为抵消 Ang II 有害作用的关键调节因子。我们最近证明，通过 AAV 介导的视网膜基因传递，保护轴的两个关键成员 ACE2 和 Ang1-7 的表达增加，可减少糖尿病引起的视网膜病理，证实增强 ACE2/Ang1-7/MAS 轴是治疗 DR 患者的一种有前途的方法。此外，增强 Ang1-7 和 ACE2 还可以改善脂质和葡萄糖代谢，改善胰岛素抵抗，并针对多种病理状况提供保护，包括糖尿病引起的肾病和心血管功能障碍。因此，理想的策略是在靶组织处全身和局部增强 ACE2/Ang1-7。该提案利用了基于使用植物叶绿体作为高效生物反应器的创新概念，用于生产和口服递送生物封装在植物细胞中的治疗性蛋白质。该治疗蛋白与舞蹈病毒素亚基 B​​ 融合，不仅促进跨粘膜转运进入循环，而且还通过 GM1 受体介导的内吞作用增强组织细胞的摄取。我们最近表明，口服治疗蛋白广泛分布在包括大脑和视网膜在内的器官中，跨越血脑/视网膜屏障；口服生物封装髓磷脂碱性蛋白可改善晚期人类和小鼠阿尔茨海默病大脑和视网膜的淀粉样蛋白负担，防止视网膜神经节细胞损失。因此，这一新颖的概念提供了一种更有效、更低成本的生产和对患者友好的治疗性蛋白质递送，特别是对中枢神经系统的递送，这长期以来一直是一个重大挑战。该提案的目标是评估在合适的动物模型中口服生物封装在植物细胞中的 Ang1-7 和 ACE2 在预防和治疗 DR 中的功效，并生成和表征可食用生菜中的转质体 ACE2 和 Ang1-7 表达，以推进生物封装蛋白递送方法的临床研究。这些拟议研究的结果将为使用基于植物叶绿体的治疗性蛋白口服递送的 DR 动物模型的可行性和有效性提供概念验证，并为启动通过口服递送生物封装 ACE2 和 Ang1-7 预防和治疗 DR 的临床试验奠定基础。