Anti-catabolic drug anchored cationic exosomes for cartilage targeting and repair

用于软骨靶向和修复的抗分解代谢药物锚定的阳离子外泌体

• 批准号: 9809789
• 负责人: Ambika Goel Bajpayee
• 金额: $ 19.63万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9809789
• 关键词: Antibodies; Arthralgia; Arthritis; Binding; Biochemical; Biodistribution; Biological; Biological Assay; Biological Transport; Cartilage; Cations; Cells; Charge; Chondrocytes; Chronic; Clinical; Coculture Techniques; Complex; Custom; Degenerative polyarthritis; Development; Diffuse; Dose; Drug Carriers; Drug Delivery Systems; Drug Transport; Electrostatics; Encapsulated; Engineering; Escherichia coli; Exhibits; Eye; Fibroblasts; Gene Expression; Genetic Engineering; Genetic Materials; Histology; Hybrids; Hydrophobicity; Inflammation; Inflammatory; Injections; Injury; Interleukin-1; Intervertebral disc structure; Intra-Articular Injections; Joints; Kinetics; Knee; Lipid Bilayers; Lipids; Mediating; Meniscus structure of joint; Mesenchymal Stem Cells; Modeling; One-Step dentin bonding system; Penetration; Peptides; Pharmaceutical Preparations; Process; Property; Proteins; Rattus; Recombinants; Role; Signal Transduction; Synovial Cell; Synovial Membrane; Tail; Therapeutic; Thick; Time; Tissues; Toxic effect; Translating; Treatment Efficacy; anakinra; base; biomaterial compatibility; bone; cartilage degradation; cartilage repair; cell motility; cell type; clinical translation; cytokine; density; design; engineered exosomes; exosome; in vivo; inhibitor/antagonist; injured; microCT; preservation; receptor; regenerative; repaired; residence; success; uptake

Project Summary Osteoarthritis (OA) is associated with severe joint pain, inflammation, and chronic cartilage degeneration. Mesenchymal stem cells (MSCs) derived exosomes are emerging as promising therapeutics for OA as they carry proteins and genetic materials that induce regenerative processes like cell migration, proliferation, differentiation and matrix synthesis. Their role in biological and transport crosstalk across multiple joint tissues and cell types, however, remains unclear. Additionally, the negative charge of exosome lipid bilayer hinders their penetration into the negatively charged cartilage. The high negative fixed charge density of cartilage offers a unique opportunity to utilize electrostatic interactions to enhance intra-tissue transport, uptake, and retention of exosomes by making them positively charged. We have designed an amphipathic cartilage penetrating cationic peptide (CP) that can rapidly diffuse through full tissue thickness due to their optimal charge, be up-taken by cells, and bind within for extended periods in both healthy and arthritic cartilage. This project will engineer cartilage targeting MSC-exosomes anchored with CPs and with an anti-catabolic OA biologic, IL-1Ra (IL-1 inhibitor) in optimal concentrations. Currently, extensive genetic engineering approaches are used to produce customized exosomes encapsulating biologics, which may compromise their intrinsic composition making their clinical translation complex. The project will use a simple one-step synthesis of grafting CP and IL-1Ra on exosome lipid bilayer. CP-exosomes can thus use cartilage as a drug depot and target cells thereby enhancing the availability of optimally loaded IL-1Ra to its receptors while preserving their intrinsic therapeutic potential. Aim 1 will engineer CP grafted MSC-exosome (CP-Exo) and characterize its intra-cartilage transport properties in healthy and arthritic states. Their transport crosstalk and uptake across multiple cell types using cytokine challenged chondrocyte and synovial cell co-cultures will be studied to understand whether their therapeutic benefits arise from cartilage or synovium targeting or both. Aim 2 will synthesize recombinant lipid fused IL-1Ra that will be anchored in different densities on exosome bilayer to form a hybrid vehicle, IL-1Ra-CP-Exo. Its bioactivity will be evaluated using cytokine challenged cartilage-synovium explant co-cultures and compared with free IL-1Ra and unmodified exosomes. Aim 3 will characterize joint kinetics, intra-cartilage uptake and biodistribution of CP-Exo in healthy and injured rat knees, and bio efficacy of IL1-Ra-CP-Exo in suppressing injury induced catabolic signaling will be evaluated using rat models of post traumatic OA. The project paves way for utilizing the intrinsic therapeutic potential of exosomes for cartilage repair as well as for its customizable development as a drug carrier allowing for adjustable intra-cartilage transport properties, easy drug anchoring and controllable loading of a variety of pro-chondrogenic protein drugs and antibodies. The success of this project can enable rapid clinical translation of exosomes as a cell-free, non-immunogenic platform for drug delivery to cartilage and other negatively charged tissues like meniscus, intervertebral discs, eye etc.

项目摘要 骨关节炎（OA）与严重的关节疼痛，炎症和慢性软骨变性有关。 间充质干细胞（MSC）衍生的外泌体在OA携带时正在成为有希望的治疗剂 诱导细胞迁移，增殖，分化等再生过程的蛋白质和遗传材料 和基质合成。它们在多种关节组织和细胞类型的生物和运输串扰中的作用， 但是，尚不清楚。此外，外泌体脂质双层的负电荷阻碍了其穿透 进入负电荷的软骨。软骨的高负固定电荷密度可提供独特 利用静电相互作用来增强组织内运输，吸收和保留的机会 外泌体通过使它们积极地充电。我们设计了一种两亲的软骨穿透阳离子 肽（CP），由于其最佳电荷而可以通过完整的组织厚度迅速扩散，并通过 细胞，并在健康和关节炎软骨中长期结合。这个项目将设计 靶向MSC - 异位体的软骨，并用CPS锚定和抗代谢OA生物学IL-1RA（IL-1）（IL-1） 抑制剂）最佳浓度。目前，使用广泛的基因工程方法来生产 定制的外泌体封装生物制剂，这可能会损害其内在成分 临床翻译复合体。该项目将使用一个简单的一步合成CP和IL-1RA的合成 外泌体脂质双层。因此，CP诊断可以将软骨用作药物仓库，并可以增强目标细胞 最佳加载IL-1RA的受体可用性，同时保留其内在的治疗潜力。 AIM 1将设计CP移植的MSC-Exosos（CP-EXO），并表征其内部运输特性 在健康和关节炎状态。他们使用细胞因子在多种细胞类型的跨多种细胞类型的传输串扰和吸收 将研究受挑战的软骨细胞和滑膜细胞共培养 益处是由软骨或滑膜靶向产生的。 AIM 2将合成重组脂质融合IL-1RA 这将固定在外泌体双层上的不同密度中，形成混合动力汽车IL-1RA-CP-EXO。它是 将使用细胞因子挑战的软骨和养殖共培养评估生物活性，并将其与 游离IL-1RA和未修饰的外泌体。 AIM 3将表征关节动力学，体内摄取和 CP-EXO在健康和受伤的大鼠膝盖中的生物分布，以及IL1-RA-CP-EXO抑制的生物效能 损伤诱导的分解代谢信号将使用创伤后OA的大鼠模型进行评估。项目铺路 利用外泌体的内在治疗潜力进行软骨维修及其可定制的方法 作为药物载体的开发，允许可调节的内部运输特性，易于锚定药物 以及各种促疾病的蛋白质药物和抗体的可控加载。这个项目的成功 可以使外泌体的快速临床翻译作为无细胞的非免疫原性平台，用于递送药物 软骨和其他带负电的组织，例如弯月面，椎间盘，眼睛等。