Immunomodulation through Nanocapsule-Mediated Cytosolic Delivery of siRNA

通过纳米胶囊介导的 siRNA 胞质传递进行免疫调节

• 批准号: 9315412
• 负责人: VINCENT M. ROTELLO
• 金额: $ 19.05万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-15 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9315412
• 关键词: Anti-Inflammatory Agents; Anti-inflammatory; Arthritis; Autoimmune Diseases; Bacterial Model; Behavior; Bypass; Caliber; Cardiovascular Diseases; Cations; Cell membrane; Cells; Clinic; Cytosol; Disease; Dose; Endocytosis; Endosomes; Face; Formulation; Goals; Hydrophobicity; Image Analysis; Immune System Diseases; Immune response; Immune system; Immunology; Immunosuppression; Immunotherapy; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Lateral; Lipopolysaccharides; Liquid substance; Malignant Neoplasms; Mediating; Membrane Fusion; Messenger RNA; Methods; Modeling; Nature; Oils; Organ; Pathway interactions; Process; Property; RNA Interference; Research; Research Design; Reverse Transcriptase Polymerase Chain Reaction; Sepsis; Small Interfering RNA; Spleen; Surface; System; TNF gene; Therapeutic; Toxic effect; Translations; base; beta Actin; cancer therapy; capsule; cytokine; dosage; expectation; experimental study; immunoregulation; in vivo; insight; knock-down; macrophage; mouse model; nanocapsule; nanomaterials; nanoparticle; programs; self assembly; tool; uptake; vector

Project Summary/Abstract Immunomodulation through Nanocapsule-Mediated Cytosolic Delivery of siRNA RNA interference is a potentially powerful strategy for immunotherapy. A key barrier to this approach is the inability to effectively deliver siRNA to the cytosol: with current strategies the vast majority of siRNA remains trapped in endosomes and is ineffective. Nanoparticle-stabilized capsules (NPSCs) deliver siRNA directly to the cytosol in a membrane fusion-like process, bypassing endocytosis. We have demonstrated effective knockdown both in vitro and in vivo in the spleen, with the latter requiring significantly lower dosing than current delivery strategies. In our proposed research we will use in vitro and in vivo experiments to optimize the immunomodulatory properties of these vehicles, focusing on reducing inflammatory response by targeting the cytokine TNF-α. Our proposed program features two Aims: Aim 1: We will fabricate and optimize therapeutic siRNA-based NPSCs, focusing on maximizing cytosolar delivery efficiency, carrier capacity, and TNF-α knockdown to macrophages while minimizing toxicity and non-specific immune response. Aim 2: We will determine the efficacy of our delivery system in lipopolysaccharide- challenged mouse models of bacterial sepsis, via imaging and evaluation of anti- inflammatory effects following siRNA-bearing NPSC treatment. The goal of this proposal is to demonstrate the utility of the NPSC platform for immunomodulation. We will build upon the highly efficient cytosolar delivery of siRNA observed in our preliminary NPSC results, evaluating and optimizing their in vivo behavior. These studies will provide critical insights to the translational potential of this vehicle, providing essential preliminary results for applications in specific immune disorders.

项目总结/摘要 通过纳米囊介导的细胞溶质递送的免疫调节 siRNA RNA干扰是一种潜在的强大的免疫治疗策略。一个关键障碍 这种方法的缺点是不能有效地将siRNA递送到胞质溶胶： 然而，绝大多数siRNA仍然被困在内体中并且是无效的。 纳米颗粒稳定的胶囊（NPSC）将siRNA直接递送到细胞质中， 膜融合样过程，绕过内吞作用。我们已经证明了 在体外和体内脾中的敲低，后者需要显著 剂量低于当前的递送策略。在我们的研究中，我们将使用 体外和体内实验，以优化这些免疫调节特性， 媒介物，专注于通过靶向细胞因子TNF-α来减少炎症反应。 我们提出的方案有两个目标： 目的1：我们将制造和优化基于siRNA的治疗性NPSC，重点是 最大化细胞太阳能递送效率、载体容量和TNF-α敲低 同时最大限度地减少毒性和非特异性免疫反应。 目的2：我们将确定我们的递送系统在脂多糖中的功效- 通过成像和评估抗细菌感染的小鼠模型， 在携带siRNA的NPSC处理后的炎症作用。 本提案的目标是展示NPSC平台的实用性， 免疫调节我们将建立在高效的细胞太阳能传递siRNA的基础上 在我们初步的NPSC结果中观察到，评估和优化其体内 行为这些研究将提供关键的见解，这种翻译潜力， 载体，为特异性免疫应用提供必要的初步结果 紊乱