Vascular Gene Delivery and Early Disease Biomarkers in Diabetic Retinopathy

糖尿病视网膜病变的血管基因传递和早期疾病生物标志物

• 批准号: 10247765
• 负责人: Daniel Mark Lipinski
• 金额: $ 36.09万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10247765
• 关键词: Abbreviations; Ablation; Address; Affect; Affinity; Age related macular degeneration; Anatomy; Antibodies; Binding; Biological Markers; Blindness; Blood; Blood Vessels; Capsid; Cardiovascular system; Catheterization; Cauterize; Cell Maintenance; Cells; Choroidal Neovascularization; Clinical Management; Clinical Trials; Degenerative Disorder; Development; Diabetic Retinopathy; Disease; Disease Progression; Dose; Early Diagnosis; Endothelial Cells; Endothelium; Etiology; Eye; Eye diseases; Florida; Functional disorder; Future; Gene Delivery; Gene Expression; Gene Transfer; Genetic Materials; Goals; Guanosine Triphosphate; Health; Heparin; Hereditary Disease; Histology; Human; Imaging Techniques; Individual; Insulin-Dependent Diabetes Mellitus; Intervention; Intracarotid; Knowledge; Laboratories; Lasers; Lead; Life; Lymphoid Tissue; Mediating; Metabolic; Methodology; Miniature Swine; Modeling; Monitor; Mutation; Nutrient; Operative Surgical Procedures; Ophthalmology; Ophthalmoscopes; Optical Coherence Tomography; Outcome; Oxygen; Pathologic; Patients; Permeability; Photoreceptors; Publications; Rattus; Recombinant adeno-associated virus (rAAV); Research; Resolution; Retina; Retinal Diseases; Retinal Neovascularization; Scanning; Specificity; Streptozocin; Structure; Sulfate; Symptoms; Techniques; Technology; Testing; Therapeutic; Therapeutic Intervention; Transgenes; Translational Research; Translations; Treatment Efficacy; Tropism; Universities; Vascular Diseases; Vascular Endothelial Cell; Vascular Endothelium; Virion; Vision; Vision research; Visual impairment; Waste Products; Wisconsin; Work; adaptive optics; adeno-associated viral vector; artery infusion; candidate identification; cellular transduction; clinical translation; clinically relevant; collaborative environment; design; diabetes management; diabetic; diabetic rat; effective therapy; fluorescence imaging; fundus imaging; gene therapy; imaging modality; imaging program; improved; in vivo; laser photocoagulation; medical schools; mutant; neurosensory; non-invasive imaging; novel; ocular imaging; ophthalmic artery; porcine model; prevent; response; retinal neuron; therapeutic evaluation; therapeutic gene; therapy development; vascular abnormality; vascular endothelial dysfunction; vector

ABSTRACT c The vasculature endothelium forms a selectively permeable barrier that facilitates transfer of nutrients, oxygen and waste products between the retina and the blood. Therefore, diseases affecting the structure and function of the vascular endothelium, such as diabetic retinopathy (DR) and age-related macular degeneration (AMD), have a devastating effect on the health of the retina and ultimately lead to severe visual impairment. Traditional treatment approaches focus on ameliorating disease symptoms that lead to vision loss, including retinal and choroidal neovascularization. Whilst effective, treatments such as laser photocoagulation are both invasive and destructive, requiring frequent interventions throughout the patient's lifetime, leading to the ablation of neurosensory retina as new blood vessels are cauterized. Moreover, these treatments fail to address the pathologic abnormalities within vascular endothelial cells (VECs) that underlie abnormal blood vessel function in DR. As such, they serve only to temporarily limit progression of the disease. In contrast to existing treatments, gene therapy represents an attractive therapeutic alternative, potentially allowing for the permanent correction of vascular dysfunction prior to the development of sight- threatening complications. The inability to efficiently deliver genetic material to vascular endothelial cells currently prohibits development of any gene therapy treatment aimed at preventing DR. We have recently taken the first step to overcoming this barrier by elucidating a recombinant adeno-associated virus (rAAV) vector mutant with enhanced affinity for VECs. We propose to further develop these vector technologies and optimize their surgical delivery through the following specific aims: 1) Evaluate endothelial cell transduction and maintenance of gene expression in normal and diabetic vasculature; 2) Characterize early stage biomarkers of DR progression and efficacy of endothelial cell gene therapy, and 3) Assess endothelial cell transduction following rAAV administration by selective intra-ophthalmic artery infusion (SIOAI). Utilizing a well-established rat model of type I diabetes (T1D) we anticipate the development of a strategy to effectively deliver genetic material in both normal and dysfunctional VECs. In doing so, we will utilize various advanced imaging modalities to quantin order to maximize the clinical translation of the proposed DR gene therapy, we will optimize key aspects relating to the targeted intravascular delivery of rAAV using a mini-swine model that accurately recapitulates human cardiovascular and ocular anatomy. The Ocular Gene Therapy Laboratory (OGTL) and Advanced Ocular Imaging Program (AOIP) at the Medical College of Wisconsin, together the University of Florida Department of Ophthalmology, provide the perfect collaborative environment to complete the proposed work. Finally, our proposal addresses an emerging need identified in the NEI Publication “Vision Research: Needs, Gaps, and Opportunities”: “develop novel, noninvasive imaging techniques for monitoring electrical or metabolic activity of retinal neurons in vivo, ideally at the spatial resolution of photoreceptors or better for early detection of disease and monitoring of therapeutic intervention.”

摘要c 脉管系统内皮形成促进营养物转移的选择性渗透屏障， 视网膜和血液之间的氧气和废物。因此，影响结构的疾病 和血管内皮功能，如糖尿病视网膜病变（DR）和年龄相关性黄斑病变 视网膜变性（AMD）对视网膜的健康具有破坏性影响，并最终导致严重的视网膜病变。 视力障碍传统的治疗方法侧重于改善疾病症状， 视力丧失，包括视网膜和脉络膜新生血管。虽然有效，但激光等治疗方法 光凝是侵入性和破坏性的，需要在患者的整个过程中频繁干预。 寿命，导致神经感觉视网膜的消融，因为新的血管被烧灼。此外，委员会认为， 这些治疗不能解决血管内皮细胞（VEC）内的病理异常， 因此，它们只能暂时限制DR的进展， 这种疾病与现有的治疗方法相比，基因治疗是一种有吸引力的治疗方法， 潜在地允许在视力发育之前永久矫正血管功能障碍， 危险的并发症无法有效地将遗传物质输送到血管内皮细胞， 细胞目前禁止开发任何旨在预防DR的基因治疗。 最近通过阐明重组腺相关病毒， 病毒（rAAV）载体突变体，其对VEC具有增强的亲和力。我们建议进一步开发这些载体 技术，并通过以下具体目标优化其手术递送：1）评估内皮细胞 正常和糖尿病血管系统中的细胞转导和基因表达的维持; 2）表征 DR进展的早期生物标志物和内皮细胞基因治疗的功效，以及3）评估 通过选择性眼内动脉输注给予rAAV后的内皮细胞转导 （SIOAI）。利用完善的I型糖尿病（T1 D）大鼠模型，我们预期开发一种新的糖尿病模型。 在正常和功能失调的VEC中有效递送遗传物质的策略。为此，我们将 利用各种先进的成像模式，以量化，以最大限度地提高临床翻译的 提出的DR基因治疗，我们将优化与靶向血管内递送相关的关键方面， rAAV使用微型猪模型，准确地概括了人类心血管和眼部解剖结构。 眼科基因治疗实验室（OGTL）和高级眼科成像计划（AOIP） 威斯康星州医学院和佛罗里达大学眼科系共同提供 完善的协作环境来完成建议的工作。最后，我们的建议涉及一个 NEI出版物“视觉研究：需求、差距和机会”中确定的新兴需求： “开发新的，非侵入性的成像技术，用于监测视网膜的电或代谢活动， 在体内的神经元，理想地在光感受器的空间分辨率或更好地用于疾病的早期检测， 监测治疗干预”。