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.

项目摘要/摘要 这项研究的目标是设计一种具有高效率的靶向生物纳米颗粒平台 颅内递送和神经胶质细胞靶向在药物递送和成像方面具有广泛的适用性。大量的 已经完成的工作是阐明某些细胞外小泡(Ev)的交叉能力。 内皮屏障，尤其是血脑屏障。其他研究已经证实，电动汽车展示了 对特定的组织和细胞类型有极好的趋向性。这项建议的重点是理解 某些电动汽车群体完成这些壮举的机制，并将它们设计成混合动力车 脂质体-EV给药平台。鉴于最近对电动汽车结构和功能的过多研究，它是 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，如 电动汽车规定的功能在这些亚群中是如何不同的。很可能是偏离目标的影响和 使用工程模拟技术捕获本地电动汽车功能的效率低下是由于它们的大量 异质性。我们的第一个假设是电动汽车的同质化朝着具有统一的窄尺寸范围 生物分子含量将导致更强大和更可控的药物输送平台，保持天然EV 功能还能减少非靶标毒性。我们的第二个假设是，同质化的电动汽车和 具有多种功能(即通过受体介导的细胞穿透有效地渗透血脑屏障)的脂质体将 提供融合了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 严格的工程使EVS均质(目标1)，同时进行生化分析以确定其机制 对EV颅内分娩很重要。我们将使用从神经质疏松症专家那里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、脑转移的乳腺癌细胞和其他胶质细胞 以及星形胶质细胞和神经元等神经细胞。确认对颅内递送重要的关键分子角色 这些研究将通过随后与载体电动汽车的融合来反馈到工程电动汽车(EEVS)的合成 (目标2)。对于设计的EEV产品，我们还将加入用已知物质装饰的合成脂质体 通过血脑屏障内皮层触发受体介导的跨细胞反应的配体。以提供最大的 为了有机会衡量功能性颅内分娩的效率，我们计划加载配方、标签和 用化疗有效载荷均质化EEVS，并确定药物释放情况、生物分布和 BBBS完整的健康小鼠的疗效，然后建立原位GBM模型(目标3)。建议的工作是 重要是因为它寻求消除困扰粒子到粒子变异性的高度混杂因素 电动汽车作为强有力的药物输送工具的有效应用。同质化EEVS的成功将导致 增加对其生物学功能的了解，并帮助其应用于抗击各种 目前的药物输送方式因低颅脑输送而受阻的神经性疾病。