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.

项目概要/摘要 通过纳米胶囊介导的胞浆传递进行免疫调节 siRNA RNA 干扰是免疫治疗的一个潜在强大策略。 这种方法的缺点是无法有效地将 siRNA 传递到细胞质中： 绝大多数 siRNA 仍然被困在内体中并且无效。 纳米颗粒稳定胶囊 (NPSC) 将 siRNA 直接递送至胞质溶胶中 我们已经证明了绕过内吞作用的膜融合样过程。 体外和体内脾脏中的敲低，后者需要显着 在我们提出的研究中，我们将使用比目前的给药策略更低的剂量。 体外和体内实验以优化这些药物的免疫调节特性 载体，专注于通过靶向细胞因子 TNF-α 来减少炎症反应。 我们提出的计划有两个目标： 目标 1：我们将制造和优化基于 siRNA 的治疗性 NPSC，重点关注 最大限度地提高细胞质传递效率、载体容量和 TNF-α 敲低 巨噬细胞，同时最大限度地减少毒性和非特异性免疫反应。 目标 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性脓毒症小鼠模型 携带 siRNA 的 NPSC 治疗后的炎症效应。 该提案的目标是展示 NPSC 平台的实用性 我们将基于 siRNA 的高效细胞太阳能递送。 在我们的初步 NPSC 结果中观察到，评估和优化它们的体内 这些研究将为这一行为的转化潜力提供重要的见解。 载体，为特异性免疫应用提供重要的初步结果 失调。