Natural Plasma Nano-EVs for Drug Delivery

用于药物输送的天然血浆纳米电动汽车

• 批准号: 10698612
• 负责人: Ajay Verma
• 金额: $ 27.54万
• 依托单位: FORMATION VENTURE ENGINEERING FOUNDRY INC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2025-03-14
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10698612
• 关键词: Antisense Oligonucleotides; Autologous; Binding; Biodistribution; Blood; Blood Plasma Volume; Bone Marrow; Brain; Cell Culture Techniques; Cell Line; Cells; Circulation; Clinical; Communication; Custom; Diameter; Disease model; Dose; Drug Carriers; Drug Delivery Systems; Drug Kinetics; Engineering; Half-Life; Human; Immunoassay; Intravenous; Ion-Exchange Chromatography Procedure; Lipids; Liposomes; Liver; Measures; Membrane; Methods; Molecular Profiling; Molecular Sieve Chromatography; Organ; Penetration; Pharmaceutical Preparations; Phase; Plasma; Positron-Emission Tomography; Procedures; Production; Property; Proteins; RNA; RNA marker; Radiolabeled; Reporting; Reproducibility; Research; Safety; Small Business Innovation Research Grant; Spleen; Surface; Testing; Therapeutic; Tissues; Tropism; cell immortalization; cell type; delivery vehicle; experience; extracellular vesicles; fluorescence imaging; human stem cells; immunogenicity; improved; in vivo; multiple omics; nano; nanoparticle; novel; pharmacologic; protein biomarkers; scale up; targeted delivery

ABSTRACT Targeted delivery of efficacious drug levels with minimal off-target effects remains a major pharmacological challenge as exemplified by the poor delivery of proteins, antisense oligonucleotides (ASO), and other therapeutics to may organs. Drug delivery platforms that allow for controlled pharmacokinetic profile and cell targeting, including nanoparticles, liposomes and cell-derived extracellular vesicles (EV), are thus aggressively pursued by biopharma. However, several key gaps remain in engineering the appropriate size and surface composition of drug carrier nanoparticles, which dictates their biodistribution. Despite their favorable immunogenicity profile, EVs generated from human stem cells or immortalized cell lines invariably show entrapment by reticuloendothelial (RES) cells in liver, spleen, and bone marrow following intravenous (IV) dosing. In contrast, the persistently high plasma level of endogenous, organ derived EVs likely reflects reduced clearance and longer circulation times, both properties important for allowing for efficient payload delivery to multiple target tissues. Also, surface molecular signatures comprised of exposed proteins and lipids, which vary across plasma EV subsets, are likely to dictate their tropism to specific organs or cells. This research will capitalize on the naturally engineered properties of endogenous EVs to develop the first human plasma derived drug delivery product. We have developed a novel and sensitive multiplexed immunoassay, suitable for the use with unprocessed plasma, to characterize plasma EV subsets based on specific profiles of surface-exposed proteins. This method enabled us to discover of a novel endogenous subset of nano-EVs (nEV), whose small diameter (10-40 nm) contrasts sharply with the 50-200 nm diameter reported for most EVs. The nEVs contain protein and RNA markers of cells representing several organs as well as surface lipid and protein features to reduce RES clearance. We hypothesize that this newly discovered EV subset is naturally adapted for long-range inter- organ communications, and thus ideally suited for drug delivery. This project will develop a robust and scalable method for isolation of large amounts of nEVs from commercially acquired human plasma. We will also quantitatively characterize in vivo nEVs PK and biodistribution using in vivo positron emission tomography (PET) and fluorescence imaging. The following specific aims will be pursued in the current proposal: Aim 1: Nano-EV production: scale-up and characterization. Aim 2: Analysis of nEV biodistribution and plasma half-life. By demonstrating that nEVs have improved biodistribution properties over traditional EVs and that they can be easily isolated in large amounts from human plasma then phase 2 of our SBIR will focus on optimizing drug loading and demonstrating efficacy in disease models.

摘要 以最小脱靶效应靶向递送有效的药物水平仍然是主要的药理学 例如，蛋白质、反义寡核苷酸（阿索）和其他寡核苷酸的不良递送是一个挑战。 对许多器官的治疗。允许受控的药代动力学特征和细胞毒性的药物递送平台 靶向，包括纳米颗粒，脂质体和细胞衍生的细胞外囊泡（EV），因此积极 被生物制药所追求。然而，在设计适当的尺寸和表面方面仍然存在几个关键差距 药物载体纳米颗粒的组成，这决定了它们的生物分布。尽管他们对 免疫原性概况，从人干细胞或永生化细胞系产生的EV总是显示出 静脉（IV）给药后肝、脾和骨髓中网状内皮（RES）细胞的截留 剂量。相比之下，内源性、器官来源的EV的持续高血浆水平可能反映了减少的内源性、器官来源的EV。 间隙和更长的循环时间，这两个特性对于允许有效的有效载荷输送到 多个靶组织。此外，表面分子特征包括暴露的蛋白质和脂质， 在血浆EV亚群中，可能决定了它们对特定器官或细胞的向性。这项研究将 利用内源性EV的天然工程特性开发第一个人血浆衍生的 药物输送产品。 我们已经开发了一种新型的和敏感的多重免疫测定，适用于未加工的 血浆，以基于表面暴露蛋白质的特异性谱表征血浆EV子集。该方法 使我们能够发现一种新的内源性纳米电动汽车（nEV），其小直径（10-40 nm） 与大多数EV报道的50-200 nm直径形成鲜明对比。nEV含有蛋白质和RNA 代表几个器官的细胞标志物以及表面脂质和蛋白质特征，以减少RES清除。 我们假设，这个新发现的EV子集自然适应于长距离的相互作用。 器官通信，因此非常适合于药物递送。 该项目将开发一种稳健且可扩展的方法，用于从商业上分离大量的nEV。 获得了人类血浆我们还将定量表征体内nEV的PK和生物分布， 体内正电子发射断层扫描（PET）和荧光成像。将努力实现以下具体目标 目标1：纳米电动汽车生产：规模扩大和特性鉴定。目的2：分析nEV 生物分布和血浆半衰期。通过证明nEV具有改善的生物分布特性， 传统的EV，它们可以很容易地从人血浆中大量分离，然后我们的第2阶段 SBIR将专注于优化药物载量并在疾病模型中证明疗效。