Homogenized, engineered extracellular vesicles for intracranial targeting

用于颅内靶向的均质化、工程化细胞外囊泡

• 批准号: 10659682
• 负责人: Randy Carney
• 金额: $ 53.36万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-04 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659682
• 关键词: Address; Astrocytes; Behavior; Biochemical; Biodistribution; Biological; Biological Assay; Biological Availability; Biological Process; Bioreactors; Blood - brain barrier anatomy; Brain; Breast Cancer Cell; Cancer cell line; Cell Line; Cells; Chemicals; Clinical; Complex; Disease; Disseminated Malignant Neoplasm; Dose; Drug Delivery Systems; Drug Targeting; Endothelium; Engineering; Exhibits; FDA approved; Failure; Feedback; Formulation; Foundations; Glioblastoma; Goals; Good Manufacturing Process; Half-Life; Heterogeneity; Hybrids; Image; Immune; Label; Libraries; Ligands; Liposomes; Malignant Neoplasms; Malignant neoplasm of brain; Measures; Mechanics; Mediating; Medicine; Methods; Microfluidics; Modeling; Molecular; Mus; Neurodegenerative Disorders; Neuroglia; Neurons; Pathway interactions; Patients; Penetration; Performance; Pharmaceutical Preparations; Phase I Clinical Trials; Population; Property; Proteins; RNA; Reporting; Research; Safety; Shapes; Solid; Source; Sterility; Structure; System; Techniques; Testing; Therapeutic; Tissues; Toxic effect; Translating; Treatment Efficacy; Tropism; Vision; Work; biomaterial compatibility; blood-brain barrier crossing; cell type; clinical practice; combat; controlled release; delivery vehicle; design; drug release profile; efficacy study; expectation; extracellular vesicles; functional improvement; immune clearance; in vivo; innovation; interest; loss of function; mimetics; mouse model; nanoarchitecture; nanocarrier; nanomaterials; nanoparticle; nanoscale; nervous system disorder; neural; next generation; novel; novel strategies; particle; personalized care; pharmacokinetics and pharmacodynamics; receptor; safety study; safety testing; success; synthetic drug; temozolomide; tool; transcytosis; tumor; uptake

PROJECT SUMMARY/ABSTRACT The objective of the proposed research is to engineer a targeted biological nanoparticle platform with high intracranial delivery and glial cell targeting for broad applicability in drug delivery and imaging. A great deal of work has already been accomplished elucidating the ability of certain extracellular vesicles (EVs) to cross endothelial barriers, especially the blood-brain barrier (BBB). Other work has established that EVs exhibit excellent tropism towards particular tissues and cell types. The focus of this proposal is to understand the mechanisms by which certain EV subpopulations accomplish these feats, and to engineer them into a hybrid liposome-EV drug delivery platform. Given the plethora of recent research into EV structure and function, it is well known that they exhibit considerable compositional heterogeneity. But fundamental questions still exist as to how EV prescribed functions differ across these subpopulations. It is likely that off-target effects and inefficiencies in capturing native EV functions with engineered mimetics are due to their substantial heterogeneity. Our first hypothesis is that homogenization of EVs towards a narrow size range with uniform biomolecular content will result in a more potent and controllable drug delivery platform that maintains native EV function yet reduces off-target toxicity. Our second hypothesis is that fusion of homogenized EVs and liposomes with various functions (i.e., efficient BBB permeation through receptor mediated transcytosis) will deliver an engineered product combining desired functions. We plan on addressing these hypotheses through rigorous engineering to homogenize EVs (Aim 1) alongside biochemical assays to detangle the mechanisms important for EV intracranial delivery. We will utilize EVs isolated from gliatropic “experts”, namely a vast library of glioblastoma (GBM) patient derived primary cell lines, brain-metastasizing breast cancer cells, and other glial and neuronal cells like astrocytes and neurons. Key molecular players important for intracranial delivery identified from those studies will feedback into synthesis of engineered EVs (eEVs) via subsequent fusion with carrier EVs (Aim 2). For the engineered eEV product, we will also incorporate synthetic liposomes decorated with known ligands to trigger receptor mediated transcytosis through the BBB endothelial layer. To provide the greatest opportunity to measure efficiency of functional intracranial delivery, we plan to load formulated, labeled, and homogenized eEVs with a chemotherapeutic payload and determine drug-release profile, biodistribution, and efficacy in healthy mice with intact BBBs and then an orthotopic GBM model (Aim 3). The proposed work is important because it seeks to eliminate the highly confounding factor of particle-to-particle variability plaguing effective application of EVs as potent drug-delivery vehicles. Success in homogenizing eEVs will result in an increased understanding of their biological function and assist in their application to combat a wide variety of neurological disorders where current drug delivery approaches are thwarted by low intracranial delivery.

项目概要/摘要 该研究的目的是设计一个具有高靶向性的生物纳米颗粒平台。 颅内递送和神经胶质细胞靶向在药物递送和成像中具有广泛的适用性。大量的 阐明某些细胞外囊泡 (EV) 交叉能力的工作已经完成 内皮屏障，特别是血脑屏障（BBB）。其他工作已证实电动汽车表现出 对特定组织和细胞类型具有良好的趋向性。该提案的重点是了解 某些电动汽车亚群实现这些壮举的机制，并将它们设计成混合动力车 脂质体-EV药物递送平台。鉴于最近对电动汽车结构和功能的大量研究， 众所周知，它们表现出相当大的成分异质性。但基本问题仍然存在 EV 指定功能在这些亚人群中有何不同。脱靶效应和 使用工程模拟物捕获原生电动汽车功能效率低下的原因是它们的大量 异质性。我们的第一个假设是，电动汽车的同质化朝着狭窄的尺寸范围发展，并且具有统一的 生物分子内容将产生更有效、更可控的药物输送平台，以维持天然 EV 功能同时降低脱靶毒性。我们的第二个假设是同质化电动汽车和汽车的融合 具有各种功能的脂质体（即通过受体介导的转胞吞作用进行有效的 BBB 渗透）将 提供结合所需功能的工程产品。我们计划通过以下方式解决这些假设 严格的工程设计使 EV 均质化（目标 1），同时进行生化分析以理清机制 对于 EV 颅内输送很重要。我们将利用从神经胶质细胞“专家”中分离出来的电动汽车，即一个巨大的图书馆 胶质母细胞瘤 (GBM) 患者来源的原代细胞系、脑转移性乳腺癌细胞和其他神经胶质细胞 以及神经元细胞，如星形胶质细胞和神经元。确定了对颅内递送重要的关键分子参与者 这些研究的成果将通过随后与载体电动汽车的融合反馈到工程电动汽车（eEV）的合成中 （目标 2）。对于工程化的 eEV 产品，我们还将结合用已知的装饰物装饰的合成脂质体 配体通过 BBB 内皮层触发受体介导的转胞吞作用。为提供最大的 有机会测量功能性颅内输送的效率，我们计划加载配制好的、贴标签的和 具有化疗有效负载的均质化 eEV，并确定药物释放曲线、生物分布和 在具有完整 BBB 的健康小鼠和原位 GBM 模型中的疗效（目标 3）。拟议的工作是 很重要，因为它试图消除困扰粒子间变异性的高度混杂因素 EV 作为有效的药物输送工具的有效应用。电动汽车同质化的成功将导致 增加对其生物学功能的了解，并协助其应用来对抗多种疾病 神经系统疾病，目前的药物输送方法因颅内输送量低而受阻。