Development of Anti-Fouling Peptide-Nanoparticle Conjugates for the Delivery of siRNA to Fractures

开发用于将 siRNA 递送至骨折的抗污肽-纳米颗粒缀合物

• 批准号: 9907178
• 负责人: Clyde Thomas Overby
• 金额: $ 4.55万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9907178
• 关键词: Adsorption; Amino Acids; Antibodies; Betaine; Biomimetics; Blood Circulation; Bone callus; Carbodiimides; Cations; Cells; Characteristics; Charge; Chemicals; Chemistry; Contralateral; Degenerative polyarthritis; Development; Disease; Drug Delivery Systems; Drug Kinetics; Dyes; Femoral Fractures; Formulation; Foundations; Fracture; Fracture Healing; Future; Half-Life; Heparin; Immune response; In Vitro; Inbred BALB C Mice; Injections; Kidney; Label; Lead; Libraries; Limb structure; Liver; Liver diseases; Measurement; Measures; Mediating; Membrane Proteins; Mesenchymal Stem Cells; Microscopy; Modeling; Modification; Monitor; Mononuclear; Mus; Musculoskeletal; Musculoskeletal System; Osteoporosis; Pathway interactions; Peptides; Peptidylprolyl Isomerase; Phagocytes; Pharmaceutical Preparations; Polymers; Property; Proteins; RNA delivery; Radial; Randomized; Research; Reverse Transcriptase Polymerase Chain Reaction; Serum; Serum Proteins; Small Interfering RNA; Surface; System; Tendinopathy; Testing; Therapeutic; Time; Tissues; Torsion; Treatment Efficacy; base; bone; bone healing; design; ethylene glycol; extracellular; healing; immunogenicity; improved; in vivo; inhibitor/antagonist; knock-down; light scattering; macrophage; nanoparticle; nanoparticle delivery; novel strategies; prediction algorithm; prevent; protein aggregation; protein aminoacid sequence; therapeutic nanoparticles; trafficking; ubiquitin-protein ligase; uptake

Small interfering RNA (siRNA) are a promising class of drugs that are limited by delivery challenges. Naked siRNA has poor cellular uptake and rapid degradation in vivo, thus siRNA is typically administered in nanoparticle (NP) formulations. To circumvent extracellular barriers of degradation and poor uptake as well as intracellular lysosomal trafficking, our lab pioneered the development of pH-responsive cationic diblock tercopolymer nanoparticles (pHCNPs) for the delivery of siRNA, which greatly improve siRNA therapeutic efficacy. Local in vivo delivery of siRNA-pHCNPs has shown promise in bone and other tissues, but many diseases, including many of the musculoskeletal system require systemic administration. Systemic delivery of NPs is hampered due to serum adsorption, leading to mononuclear phagocyte system (MPS) uptake and poor circulation times, and may potentiate immunogenicity. In fact, many siRNA-NP therapeutics that are approved or in late stages of development take advantage of MPS accumulation in liver, targeting hepatic diseases. Reducing protein-NP interactions is key to improving the systemic delivery of siRNA-NPs to reach other target tissues. Poly(ethylene glycol) (PEG) modification (PEGylation) of NPs is the current standard to reduce protein adsorption, but it also reduces NP efficacy by hampering uptake and may induce immunological responses due to anti-PEG antibodies. Zwitterionic (ZI) moieties have shown great promise in reducing protein adsorption and are less disruptive to NP functional characteristics than PEG. Biomimetic ZI peptides (ZIPs) are a promising new approach to improve polymeric NP pharmacokinetic properties. In particular, semi-randomized ZIPs (srZIPs) allow testing of charge sequence semi-independently of amino acid composition. These attributes altogether lead to the hypothesis that semi-randomized ZIPs (srZIPs) designed with low aggregation potential can be used to modify NPs to improve systemic circulation and siRNA delivery by reducing NP-serum protein interactions. We will test this hypothesis in three ways. In Aim 1, we will generate a library of srZIP-pHCNP conjugates to test the effects on serum-induced aggregation compared to naïve pHCNPs and PEG-pHCNPs. In Aim 2A, we will test these conjugates in vitro using target cells (mesenchymal stem cells) and in MPS cells (macrophages), and in Aim 2B we will evaluate improvements in circulation times of the pHCNP conjugates in vivo. In Aim 3 we will use an established mouse femur fracture model to investigate fracture accumulation of pHCNP bearing therapeutic anti-WWP1 siRNA (WW Domain Containing E3 Ubiquitin Protein Ligase 1, a negative regulator of fracture healing), which we have shown to expedite bone fracture healing in using a local delivery approach. At the completion of this project, we expect to identify new peptide-based anti-aggregation approaches for the delivery of siRNA in polymeric nanoparticles, laying the foundation for future systemic delivery of siRNA for musculoskeletal applications and beyond.

小干扰 RNA (siRNA) 是一类很有前途的药物，但受到递送挑战的限制。裸 siRNA 的细胞摄取较差且体内降解较快，因此 siRNA 通常在体内施用 纳米颗粒（NP）配方。规避细胞外降解障碍和吸收不良以及 细胞内溶酶体运输，我们的实验室率先开发了 pH 响应型阳离子二嵌段 用于递送 siRNA 的三元共聚物纳米粒子 (pHCNP)，极大地改善了 siRNA 的治疗效果 功效。 siRNA-pHCNP 的局部体内递送已在骨骼和其他组织中显示出前景，但许多 包括许多肌肉骨骼系统疾病需要全身给药。系统性交付 纳米颗粒因血清吸附而受到阻碍，导致单核吞噬细胞系统 (MPS) 摄取不良 循环时间，并可能增强免疫原性。事实上，许多已获批准的 siRNA-NP 疗法 或者在发育后期利用 MPS 在肝脏中的积累，针对肝脏疾病。 减少蛋白质-纳米颗粒相互作用是改善 siRNA-纳米颗粒系统递送以到达其他靶标的关键 组织。 NP 的聚乙二醇 (PEG) 修饰（聚乙二醇化）是目前减少蛋白质含量的标准 吸附，但它也会通过阻碍吸收来降低 NP 功效，并可能诱导免疫反应 由于抗 PEG 抗体。两性离子 (ZI) 部分在减少蛋白质吸附方面显示出巨大的前景 与 PEG 相比，对 NP 功能特性的破坏较小。仿生 ZI 肽 (ZIP) 是一种 改善聚合物纳米粒子药代动力学特性的有前途的新方法。特别是半随机 ZIP (srZIP) 允许半独立于氨基酸组成测试电荷序列。这些 属性共同导致了这样的假设：半随机 ZIP（srZIP）设计具有低 聚集潜力可用于修饰 NP，以改善体循环和 siRNA 递送 通过减少 NP-血清蛋白相互作用。我们将以三种方式检验这个假设。在目标 1 中，我们将 生成 srZIP-pHCNP 缀合物库，以测试与 天然 pHCNP 和 PEG-pHCNP。在目标 2A 中，我们将使用靶细胞在体外测试这些缀合物 （间充质干细胞）和 MPS 细胞（巨噬细胞），在 Aim 2B 中，我们将评估以下方面的改进： pHCNP 缀合物在体内的循环时间。在目标 3 中，我们将使用已确定的小鼠股骨骨折 用于研究带有治疗性抗WWP1 siRNA（WW域）的pHCNP的断裂累积的模型 含有 E3 泛素蛋白连接酶 1（骨折愈合的负调节因子），我们已证明它可以 使用局部输送方法加速骨折愈合。在这个项目完成后，我们预计 确定新的基于肽的抗聚集方法，用于在聚合物中递送 siRNA 纳米颗粒，为未来肌肉骨骼应用中 siRNA 的系统递送奠定了基础 超过。