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: 10176484
• 负责人: Ambika Goel Bajpayee
• 金额: $ 23.55万
• 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 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10176484
• 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; 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; regeneration potential; regenerative; repaired; residence; stem cell exosomes; 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.

项目概要 骨关节炎 (OA) 与严重的关节疼痛、炎症和慢性软骨退化有关。 间充质干细胞 (MSC) 衍生的外泌体正在成为治疗 OA 的有前途的疗法，因为它们携带 诱导细胞迁移、增殖、分化等再生过程的蛋白质和遗传物质 和矩阵合成。它们在跨多种关节组织和细胞类型的生物和运输串扰中的作用， 然而，目前尚不清楚。此外，外泌体脂质双层的负电荷阻碍了它们的渗透 进入带负电的软骨。软骨的高负固定电荷密度提供了独特的 有机会利用静电相互作用来增强组织内的运输、摄取和保留 外泌体通过使它们带正电荷。我们设计了一种两亲性软骨穿透阳离子 由于其最佳电荷，肽（CP）可以快速扩散到整个组织厚度，被 细胞，并在健康软骨和关节炎软骨中长时间结合。该项目将设计 软骨靶向 MSC 外泌体锚定于 CP 和抗分解代谢 OA 生物制剂 IL-1Ra (IL-1 抑制剂）在最佳浓度。目前，广泛使用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，使其 临床翻译复杂。该项目将使用一种简单的一步合成方法将 CP 和 IL-1Ra 接枝到 外泌体脂质双层。因此，CP-外泌体可以使用软骨作为药物库和靶细胞，从而增强 最佳负载的 IL-1Ra 对其受体的可用性，同时保留其内在的治疗潜力。 目标 1 将设计 CP 移植的 MSC-外泌体 (CP-Exo) 并表征其软骨内运输特性 在健康和关节炎状态下。使用细胞因子，它们的运输串扰和跨多种细胞类型的摄取 将研究受挑战的软骨细胞和滑膜细胞共培养物，以了解它们的治疗作用是否 益处来自软骨或滑膜靶向或两者。目标2将合成重组脂质融合IL-1Ra 它将以不同的密度固定在外泌体双层上，形成混合载体 IL-1Ra-CP-Exo。它是 将使用细胞因子激发的软骨-滑膜外植体共培养物评估生物活性，并与 游离 IL-1Ra 和未修饰的外泌体。目标 3 将表征关节动力学、软骨内摄取和 CP-Exo 在健康和受伤大鼠膝盖中的生物分布，以及 IL1-Ra-CP-Exo 在抑制中的生物功效 将使用创伤后 OA 大鼠模型评估损伤诱导的分解代谢信号传导。该项目铺装 利用外泌体的内在治疗潜力进行软骨修复及其可定制的方法 开发为药物载体，可调节软骨内运输特性，易于药物锚定 以及多种促软骨蛋白药物和抗体的可控负载。这个项目的成功 可以将外泌体快速临床转化为无细胞、非免疫原性的药物递送平台 软骨和其他带负电的组织，如半月板、椎间盘、眼睛等。

项目成果

Sustained intra-cartilage delivery of interleukin-1 receptor antagonist using cationic peptide and protein-based carriers.

使用阳离子肽和基于蛋白质的载体持续软骨内递送白细胞介素 1 受体拮抗剂。

• DOI: 10.1016/j.joca.2023.01.573
• 发表时间: 2023
• 期刊: Osteoarthritis and cartilage
• 影响因子: 7
• 作者: Mehta,S;Boyer,TL;Akhtar,S;He,T;Zhang,C;Vedadghavami,A;Bajpayee,AG
• 通讯作者: Bajpayee,AG

Effects of polycationic drug carriers on the electromechanical and swelling properties of cartilage

聚阳离子药物载体对软骨机电和溶胀性能的影响

• DOI: 10.1016/j.bpj.2022.06.024
• 发表时间: 2022
• 期刊: Biophysical Journal
• 影响因子: 3.4
• 作者: Warren, Matthew R.;Vedadghavami, Armin;Bhagavatula, Sanjana;Bajpayee, Ambika G.
• 通讯作者: Bajpayee, Ambika G.

Milk exosomes with enhanced mucus penetrability for oral delivery of siRNA.

• DOI: 10.1039/d0bm01497d
• 发表时间: 2021-06-15
• 期刊: Biomaterials science
• 影响因子: 6.6
• 作者: Warren MR;Zhang C;Vedadghavami A;Bokvist K;Dhal PK;Bajpayee AG
• 通讯作者: Bajpayee AG