Enhanced intratympanic delivery of therapeutics to treat and prevent hearing loss using nanovesicles in the porcine model

在猪模型中使用纳米囊泡增强治疗剂的鼓室内递送以治疗和预防听力损失

基本信息

批准号：
10665079
负责人：
Adele Moatti
金额：
$ 10.23万
依托单位：
NORTH CAROLINA STATE UNIVERSITY RALEIGH
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-15 至 2024-06-30
项目状态：
已结题

项目摘要

PROJECT SUMMARY My long-term goal is to help develop and deliver regeneration therapeutics useful for the clinical treatment of hearing loss. Unfortunately, at this time there is no high throughput, non-destructive method of therapeutic delivery into the cochlea/inner ear. The objectives of this proposal, the next step toward the attainment of this long-term goal, are to use a relevant large animal model (the pig) to i) develop an ex-vivo model system, which mimics human round window membrane (RWM), to track the passage of therapy-related substances through the RWM, and ii) create a method to enhance the intratympanic delivery of therapeutics into the inner ear using nanovesicles including extracellular vesicles (exosomes). The central hypothesis, supported by our preliminary data, is that an ex-vivo porcine RWM model can be used to determine optimal conditions for the transport of exosomes (or other nanovesicles) to deliver cargo through the RWM into the inner ear, thereby facilitating the high-efficiency delivery of therapeutics. The rationale for this proposal is that its successful completion is likely to offer a framework whereby a new pool of therapeutics can be tested for non-surgical delivery into the inner ear, in a large animal model that has the preclinical advantage over rodents in terms of size, physiology, and genetic similarity (amino-acid sequences of common deafness genes) to humans. The following specific aims will be pursued during K99 (aim 1 and 2) and R00 phase: Aim 1) To further validate the ex-vivo porcine RWM model to demonstrate drug permeability equivalent to that reported for human tissue; Aim 2) To identify and evaluate exosomes and other nanovesicles enhancing cargo transport across porcine RWM in-vitro; Aim 3) To evaluate in-vivo transport across porcine RWM by nanovesicles. Under the first aim, we will measure the permeability of therapeutics with known delivery efficiencies and some promising therapy related materials using a preliminary viability-verified ex-vivo RWM model and compare those with values reported for human tissue. For the second aim, nanovesicles including: RWM exosomes (successfully isolated and characterized as a preliminary result), mesenchymal stem cell exosomes (gold standard), and liposomes (FDA approved), that are loaded with promising therapeutics will be evaluated for the efficiency of transport using the ex-vivo RWM model. Finally, for the last aim, once the parameters associated with optimal transport through the RWM are established with the ex-vivo model, we will inject therapeutic-loaded nanovesicles through the porcine tympanic membrane and measure their passage across RWM and uptake by cochlear cells in vivo. Upon completion of the K99, the expected outcomes are 1. Availability of a safe and translatable platform to test transport of therapeutics into the inner ear, and 2. Data on the efficiency of nanovesicles as novel nonsurgical transport of promising therapeutics to the ear. These results are expected to have a positive impact because they could improve drug screening for delivery and is likely to boost delivering novel regenerative therapeutics to treat and prevent hearing loss.

项目概要我的长期目标是帮助开发和提供可用于临床治疗的再生疗法听力损失。不幸的是，目前还没有高通量、非破坏性的治疗方法。输送到耳蜗/内耳。本提案的目标、实现这一目标的下一步长期目标是使用相关的大型动物模型（猪）来 i) 开发离体模型系统，该系统模仿人类圆窗膜（RWM），追踪治疗相关物质通过 RWM，以及 ii) 创建一种方法，使用以下方法增强鼓室内治疗药物输送至内耳：纳米囊泡，包括细胞外囊泡（外泌体）。我们初步支持的中心假设数据表明，离体猪 RWM 模型可用于确定运输的最佳条件外泌体（或其他纳米囊泡）通过 RWM 将货物输送到内耳，从而促进高效率地提供治疗。该提案的理由是它很可能成功完成提供一个框架，可以测试新的治疗方法是否通过非手术方式输送到体内在大型动物模型中，耳朵在大小、生理学和功能方面比啮齿类动物具有临床前优势与人类的遗传相似性（常见耳聋基因的氨基酸序列）。具体目标如下将在 K99（目标 1 和 2）和 R00 阶段进行：目标 1) 进一步验证离体猪 RWM 证明药物渗透性与人体组织报告的药物渗透性相当的模型；目标 2) 识别并评估外泌体和其他纳米囊泡，增强猪 RWM 体外的货物运输；目标 3) 至评估纳米囊泡跨猪 RWM 的体内运输。在第一个目标下，我们将衡量具有已知递送效率的治疗剂的渗透性和一些有前景的治疗相关材料初步可行性验证的离体 RWM 模型，并将其与报告的人体组织值进行比较。对于第二个目标，纳米囊泡包括：RWM 外泌体（成功分离并表征为初步结果）、间充质干细胞外泌体（金标准）和脂质体（FDA 批准），将使用离体 RWM 模型评估装载有前景的治疗药物的运输效率。最后，对于最后一个目标，一旦建立了与通过 RWM 的最佳运输相关的参数对于离体模型，我们将通过猪鼓膜注射载有治疗药物的纳米囊泡并测量它们穿过 RWM 的通道以及体内耳蜗细胞的摄取。完成 K99 后，预期成果是 1. 提供一个安全且可翻译的平台来测试治疗药物的运输内耳，以及 2. 纳米囊泡作为新型非手术运输有前景的治疗方法的效率数据到耳朵。这些结果预计将产生积极影响，因为它们可以改善药物筛选交付，并可能促进新型再生疗法的交付，以治疗和预防听力损失。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Tissue clearing and three-dimensional imaging of the whole cochlea and vestibular system from multiple large-animal models.

多个大型动物模型的整个耳蜗和前庭系统的组织透明化和三维成像。

DOI：
发表时间：
2023-04-13
期刊：
影响因子：
0
作者：
Moatti, Adele;Cai, Yuheng;Li, Chen;Popowski, Kristen D;Cheng, Ke;Ligler, Frances S;Greenbaum, Alon
通讯作者：
Greenbaum, Alon