Identification of eye-homing peptides and their use for targeted liposomal drug delivery in posterior uveitis

眼归巢肽的鉴定及其在后葡萄膜炎靶向脂质体药物递送中的应用

• 批准号: 10612913
• 负责人: KAMAL D MOUDGIL
• 金额: $ 19.31万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10612913
• 关键词: Address; Adjuvant Arthritis; Adverse effects; Affect; Alzheimer' s Disease; Amino Acids; Animal Cancer Model; Animals; Antibodies; Antigens; Arthritis; Attention; Autoantigens; Autoimmune; Autoimmune Diseases; Autoimmunity; Bacteriophages; Binding; Biological Availability; Biology; Blindness; Blood Vessels; Blood-Retinal Barrier; Cells; Characteristics; Choroid; Chronic Disease; Collaborations; Cyclic Peptides; Cysteine; Disease; Drug Delivery Systems; Drug Targeting; Encapsulated; Endothelium; Experimental Autoimmune Encephalomyelitis; Eye; Eye diseases; Gene Expression Profiling; Hemorrhage; Heterogeneity; Home; Homing; Human; Hypoxia; Immune; Immunization; Individual; Inflammation; Inflammation Mediators; Joints; Lesion; Libraries; Ligands; Liposomes; Lymphoid Tissue; Mediator; Methodology; Methods; Methotrexate; Modeling; Molecular; Molecular Profiling; Multiple Sclerosis; Neural Retina; Organ; Oxygen; Pathogenesis; Pathogenicity; Pathology; Peptide Library; Peptide Phage Display Library; Peptides; Peripheral; Pharmaceutical Preparations; Pharmacodynamics; Posterior Uveitis; Procedures; Process; Proteins; Publishing; Rattus; Reporting; Retina; Retinal Detachment; Retinal Diseases; Rheumatoid Arthritis; Rodent; Site; Specificity; T-Lymphocyte; Testing; Tissues; Topical application; Toxic effect; Uvea; Uveitis; Vascular Endothelium; Visual impairment; Work; anterior chamber; autoimmune pathogenesis; autoimmune uveitis; autoreactive T cell; design; human model; improved; in vivo; innovation; interstitial retinol-binding protein; intravitreal injection; medication safety; migration; molecular marker; nanoparticle; neoplastic cell; novel; novel therapeutics; posterior eyeball chamber; protein aminoacid sequence; screening; systemic toxicity; tool; trafficking; tumor

Human posterior segment uveitis of autoimmune origin is a chronic disease that can impair vision and result in blindness. A variety of autoantigens, including interphotoreceptor retinoid binding protein (IRBP), have been implicated in uveitis pathogenesis. The T cells reactive against these antigens access the eye through the blood-retinal barrier and cause local inflammation and tissue damage. Two of the main challenges in non- infectious posterior uveitis are – 1) to define the molecular changes induced during uveitis in endothelium of the choroidal and retinal vasculature, and 2) to devise novel ways to direct the systemically-administered drugs primarily into the eye to enhance their efficacy but minimize adverse effects. We hypothesize that the vascular endothelium of the eye in uveitis is characterized by unique molecular markers that facilitate both selective migration of the pathogenic T cells into the target organ and cellular interaction with the inducers/ mediators of inflammation and tissue damage. Accordingly, the identification of peptide ligands (by phage peptide library screening) that interact with such markers would be invaluable tools to study disease pathogenesis. Furthermore, these peptide ligands can be exploited for targeted drug delivery to the eye with the objective of enhancing efficacy, but reducing systemic toxicity of those drugs. In collaboration with Dr. Erkki Ruoslahti (Sanford-Burnham, La Jolla), who pioneered the concept of vascular ‘address molecules’ or ‘zip codes’, we published a study on the molecular profiling of the synovial vasculature in adjuvant arthritis in rats (PNAS 2011). We identified unique joint-homing peptide ligands using an innovative approach of ex vivo/ in vivo enrichment of clones from a phage peptide-display library. Recently, we also reported CNS-homing peptides in the EAE model of human multiple sclerosis. The advantage of using phages to detect tissue-specific markers is that there is no a priori bias in predicting the peptide sequences/motifs. And, unlike antibodies, phage peptides interact with functional domains of target molecules. We now plan to apply this methodology to rat experimental autoimmune uveitis (EAU). The aims of our study in collaboration with experts in EAU (Dr. Caspi and Dr. Sun), [in vascular biology (Dr. Teesalu from Ruoslahti group), and in pharmacodynamics (Dr. Swaan)] are: Aim 1. To identify peptide ligands homing to the diseased eye in EAU using the ex vivo/ in vivo screening of phage peptide library. (a) To identify unique peptide-encoding phages that home to the vasculature and retina of the diseased eye; and (b) to determine the relative specificity of select phages/ peptides [for other tissues within the eye as well as other organs in EAU versus control (healthy/EAE) rats.] Aim 2. To use the eye-homing peptides for targeted drug delivery to the diseased sites in EAU. To use a specific peptide to guide liposomes encapsulating a drug (e.g., methotrexate) [to the diseased sites in eyes in EAU, and compare the pharmacodynamics and safety of that drug] with that using plain liposomes or free drug. The results would advance understanding of pathogenesis of human uveitis and designing of novel therapies.

自身免疫性起源的人后段葡萄膜炎是一种慢性疾病， 导致失明。多种自身抗原，包括光感受器间类维生素A结合蛋白（IRBP）， 与葡萄膜炎发病机制有关。对这些抗原有反应的T细胞通过以下途径进入眼睛： 血视网膜屏障，并导致局部炎症和组织损伤。两个主要的挑战，在非 感染性后葡萄膜炎是：1）定义葡萄膜炎期间在内皮细胞中诱导的分子变化， 脉络膜和视网膜血管系统，以及2）设计新的方法来引导全身给药的药物 主要是进入眼睛，以提高其功效，但最大限度地减少副作用。我们假设血管 葡萄膜炎中的眼内皮的特征在于独特的分子标记物， 病原性T细胞向靶器官的迁移以及与病原性T细胞的诱导物/介导物的细胞相互作用 炎症和组织损伤。因此，肽配体的鉴定（通过噬菌体肽库 筛选）与这些标志物相互作用的基因将是研究疾病发病机制的宝贵工具。 此外，这些肽配体可用于向眼睛的靶向药物递送，目的是 增强功效，但降低这些药物的全身毒性。与Erkki Ruoslahti博士合作 （Sanford-Burnham，拉霍亚）是血管“地址分子”或“邮编”概念的先驱，我们 发表了一项关于大鼠佐剂性关节炎滑液血管系统分子谱的研究（PNAS 2011年）。我们使用体外/体内的创新方法鉴定了独特的联合归巢肽配体 从噬菌体肽展示文库富集克隆。最近，我们还报道了CNS归巢肽在 人类多发性硬化症的EAE模型。利用PCR检测组织特异性标志物的优势 在预测肽序列/基序时不存在先验偏差。与抗体不同，噬菌体 肽与靶分子的功能结构域相互作用。我们现在计划将这种方法应用于大鼠 实验性自身免疫性葡萄膜炎（EAU）。我们与EAU专家（Caspi博士）合作研究的目的是 和Sun博士），[血管生物学（Ruoslahti组的Teesalu博士）和药效学（Swaan博士）] 目标：1。使用离体/体内免疫荧光技术鉴定EAU中归巢至患病眼的肽配体， 噬菌体肽库的筛选。(a)为了鉴定独特的肽编码序列， （B）确定所选血管和视网膜的相对特异性， 肽[EAU与对照（健康/EAE）大鼠眼内其他组织以及其他器官。] 目标2.目的利用眼归巢肽靶向治疗EAU。使用 引导包封药物的脂质体的特异性肽（例如，甲氨蝶呤）[在眼睛的患病部位， EAU，并比较该药物与普通脂质体或游离药物的药效学和安全性。 本研究结果有助于加深对葡萄膜炎发病机制的认识和设计新的治疗方法。