Acoustically targeted, high-resolution, site-specific, transretinal delivery of macromolecules

声学靶向、高分辨率、位点特异性、经视网膜输送大分子

• 批准号: 10706971
• 负责人: Benjamin J Frankfort
• 金额: $ 19.05万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10706971
• 关键词: Acoustics; Acute; Affect; Age; Animal Experimentation; Animals; Blindness; Blood - brain barrier anatomy; Blood Circulation; Blood capillaries; Blood-Retinal Barrier; Brain; Cells; Clinic; Clinical; Clinical Trials; Contracts; Contrast Media; Definity; Diameter; Disease; Drug Delivery Systems; Dyes; Evans blue stain; Eye; Eye diseases; FDA approved; Focused Ultrasound; Frequencies; Gene Delivery; Gene Expression; Gene Transduction Agent; Genes; Genetic Materials; Hemorrhage; Histology; Hour; Human; Inflammation; Inherited; Injections; Intravenous; Long-Term Effects; Methodology; Methods; Microbubbles; Muller' s cell; Mus; Operative Surgical Procedures; Ophthalmologic Surgical Procedures; Outcome; Patients; Peripheral; Persons; Petechiae; Prevalence; Procedures; Process; Proteins; Protocols documentation; RPE65 protein; Reaction; Regional Disease; Reporter Genes; Resolution; Retina; Retinal Detachment; Retinal Diseases; Retinal Ganglion Cells; Risk Reduction; Rodent; Safety; Site; Specialist; Specificity; Surgeon; System; Techniques; Testing; Thick; Tight Junctions; Tissues; Tropism; Veins; Viral; Viral Vector; Virus; Work; adeno-associated viral vector; bone; cell type; cellular transduction; delivery vehicle; detector; experience; gene replacement therapy; gene therapy; improved; innovation; intravenous injection; macromolecule; nanometer; pressure; promoter; prototype; real time monitoring; retinal damage; sight restoration; skills; small molecule; stem; subretinal injection; success; therapeutic gene; transduction efficiency; ultrasound

Project summary We propose to develop a method of non-surgical, spatially precise, ultrasound-enhanced delivery of macromolecules to the retina. The need for such delivery methods stems from the prevalence of recent success in the treatment of retinal disorders by gene therapy. Inherited retinal diseases affect several million people worldwide and often result in vision loss and blindness. Most retinal disorders are incurable, but recent advances in gene therapy have restored vision and hope to many. In animal research, gene delivery to the eye has also helped uncover mechanisms of retinal disease. However, in both the clinic and animal research, delivery of genes to the eye is challenging. Currently, the delivery of genetic material requires surgery performed by a vitreo-retinal surgeon to access the subretinal space for the injection of genetic material. This process is technically difficult and can result in serious complications. Here, we will develop a new method of macromolecule and gene delivery to the retina that does not require eye surgery or intraocular injection of genetic material. In this method called, Enhanced Transretinal Ultrasound Delivery (ETUDE), we combine our experience in focused-ultrasound gene delivery and retinal disorders. In ETUDE, focused ultrasound (FUS) is targeted with high precision to a small region of retina. Then, a clinically-approved microbubble contrast agent is injected through a peripheral vein. In the brain, such a contrast agent responds to ultrasound and exerts mild pressure on interior lumen of capillaries. This pressure then opens the tight junctions in blood-brain barrier (BBB) and allows for free passage of molecules up to ~20 nanometers in diameter. This BBB opening lasts for several hours and has was previously used for delivery of small molecules, proteins, and viral vectors. The retina contains a similar vasculature referred to as a blood-retinal barrier (BRB). We hypothesize that BRB and BBB react similarly to the FUS in presence of an ultrasound contrast agent and will similarly enable site-specific delivery of molecules to the eye. To enable high-precision, high-safety gene delivery to the eye we propose to use an innovative method of spatial targeting of ultrasound. We will use high-frequency ultrasound and record the ultrasound echo of the microbubble contrast agent in the retina. We expect to target retinal ~300-micron sized regions spanning the retinal thickness in mice. Throughout this project, we will develop safe protocols for ETUDE, and quantitatively characterize its efficiency, spatial precision, and any potential tissue damage in gene delivery of intravenously applied viral vectors, and enable cell-type specific gene delivery to retinal ganglion cells (RGC). If successful, we will have enabled a safe, non-surgical, site-specific, gene and macromolecule delivery to specific retinal cell-types.

项目摘要 我们建议开发一种非手术，空间精确，超声增强的方法 大分子到视网膜。这种交付方法的需求源于最近成功的盛行 通过基因治疗治疗视网膜疾病。继承的视网膜疾病影响数百万人 在全球范围内，通常会导致视力丧失和失明。大多数视网膜疾病都是无法治愈的，但最近的进步 在基因疗法中，已经恢复了许多人的视力和希望。在动物研究中，向眼睛的基因传递也 帮助发现了视网膜疾病的机制。但是，在诊所和动物研究中 眼睛的基因具有挑战性。目前，遗传物质的递送需要A进行手术 玻璃体视网膜外科医生进入视网膜下空间以注射遗传物质。这个过程是 在技​​术上困难，可能导致严重的并发症。在这里，我们将开发一种新方法 大分子和基因递送到视网膜，不需要眼科手术或眼内注射遗传 材料。在这种称为增强的转视网膜超声输送（ETUDE）的方法中，我们结合了我们的 具有重点散发基因递送和视网膜疾病的经验。在Etude中，聚焦超声（FUS）是 以高精度针对视网膜的一小部分。然后，通过临床批准的微泡造影剂 通过外围静脉注射。在大脑中，这种对比剂对超声反应并发挥轻度 毛细血管内腔的压力。然后，这种压力打开了血脑屏障（BBB）的紧密连接 并允许直径高达约20纳米的分子自由通过。这个BBB开放持续了几个 小时并以前用于递送小分子，蛋白质和病毒载体。视网膜 包含类似的脉管系统，称为血视网膜屏障（BRB）。我们假设BRB和BBB 在存在超声对比剂的情况下，与FUS的反应类似，并将类似地启用特定地点 分子递送到眼睛。为了使高精度，高安全基因传递到眼睛，我们建议 使用一种创新的超声空间靶向方法。我们将使用高频超声并记录 视网膜中微气泡对比剂的超声回响。我们希望靶向视网膜〜300微米 大小的区域跨越小鼠的视网膜厚度。在整个项目中，我们将为 效率，空间精度和基因中任何潜在的组织损伤表征其效率，空间精度和任何潜在的组织损伤 静脉内应用病毒载体的递送，并启用细胞类型的特异性基因递送到视网膜神经节细胞 （RGC）。如果成功，我们将启用安全，非手术，特定地点，基因和大分子的递送 特定的视网膜细胞类型。