Dendritic Cell-Targeting Microparticles for Subcellularly-Targeted Delivery of In

用于亚细胞靶向递送 In 的树突状细胞靶向微粒

• 批准号: 8707719
• 负责人: Benjamin George Keselowsky
• 金额: $ 2.07万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8707719
• 关键词: Address; Antigen Targeting; Antigen-Presenting Cells; Antigens; Autoantigens; Autoimmune Diseases; Autoimmune Responses; Beta Cell; Biocompatible Materials; Biological Factors; Biological Products; Blood; Bolus Infusion; Cell Surface Receptors; Cell surface; Cells; Cholecalciferol; Data; Dendritic Cells; Development; Diabetes Mellitus; Dose; Drug Formulations; Encapsulated; Endosomes; Engineering; Environment; Generations; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Homing; ITGAX gene; Immune; Immune response; Immune system; Immunity; Immunosuppression; In Vitro; Inbred NOD Mice; Injectable; Injection of therapeutic agent; Insulin; Insulin-Dependent Diabetes Mellitus; Intravenous; Life; Location; Lymph; Maintenance; Modification; Organ; Particle Size; Particulate; Patients; Peptides; Phagocytes; Pharmaceutical Preparations; Pharmacologic Substance; Phenotype; Population; Prevention; Process; Proteins; Recruitment Activity; Regulatory T-Lymphocyte; Research; Shipping; Ships; Site; Subcutaneous Injections; Surface; System; T cell response; T-Lymphocyte; Testing; Therapeutic; Time; Tissues; Treatment Cost; Vaccination; Vaccines; Work; base; cell type; chemokine; conditioning; controlled release; economic impact; extracellular; in vivo; innovation; monocyte; novel; particle; prevent; receptor; release factor; response; targeted delivery

DESCRIPTION (provided by applicant): Type 1 diabetes (T1D) develops as a result of insufficient insulin being produced due to a self-destructive immune response against insulin producing beta cells. Although a number of factors are known to promote advantageous immune cell responses in experimental systems for T1D, systemic intravenous delivery of these agents often results in significant harmful off-target effects due to the uncontrolled dosing of bystander cells, tissues and organs. This project focuses on the targeted in vivo delivery of pro-tolerance factors and insulin antigen, in particulate form, targeted to a key immune cell type, dendritic cells (DCs). Dendritic cells, critical for maintenance and initiation of immunity to foregn antigens and tolerance to self-antigens, are phagocytic, antigen presenting cells. This makes DCs an ideal recipient for the targeted delivery of agents provided in particulate form. Moreover, exogenous conditioning of DCs with certain immunomodulatory agents has been shown to induce a pro-tolerance DC phenotype as well as ameliorate T1D. Vaccination with DC-targeting microparticles (MPs) holds promise to correct T1D autoimmune responses, critically, without the costly ex vivo manipulations required of DC-based cellular therapy. This enables the potential for widespread use. Micron-sized biodegradable polymeric particles are phagocytosable, which effectively promotes delivery of encapsulated factors to intracellular sites of DCs over non-phagocytes. These phagocytosable particles can be further targeted to DCs by surface immobilizing molecules targeting DC receptors. Larger (but still small enough to be injectable), non-phagocytosable biodegradable polymeric particles provide controlled release of encapsulated factors to the local extracellular environment at the subcutaneous injection site. Encapsulated factors in these large particles consist of bioactive factors for which DCs have the cognate cell-surface receptors. The objective of this proposal is to engineer a subcutaneously injectable dual MP vaccine system consisting of i.) Phagocytosable DC-targeting MPs delivering antigen and immunomodulatory factor (insulin and vitamin D3) to intracellular sites; and ii.) Non-phagocytosable MPs to deliver, extracellular, factors (GM- CSF and TGF-b1) for DC recruitment and tolerance induction. We expect to effect a pro-tolerogenic DC phenotype and promote induction of regulatory T-cells, suppression of auto-reactive T-cells, and prevent and reverse diabetes in non-obese diabetic (NOD) mice. We hypothesize that the combination of the multiple components in the dual MP system will more effectively provide robust, durable antigen-specific immune suppression than single-component formulations, either in MPs or in soluble form. Aim 1 is to formulate the dual MP system, test it in vitro by characterizing DC phenotype (activated, immature or tolerogenic) and T-cell response (stimulation, Th1, Th2, Treg, or Th17). Aim 2 is to evaluate the ability of the dual MP formulation in vivo, aiming to prevent and reverse diabetes in NOD mice. This novel and innovative approach holds promise for correcting autoimmune responses in T1D and represents a simple, clinically translatable system.

描述（由申请人提供）：1 型糖尿病 (T1D) 的发生是由于针对产生胰岛素的 β 细胞的自我破坏性免疫反应导致胰岛素产生不足。尽管已知许多因素可以在 T1D 实验系统中促进有利的免疫细胞反应，但由于旁观者细胞、组织和器官的剂量不受控制，这些药物的全身静脉注射通常会导致显着有害的脱靶效应。该项目的重点是在体内以颗粒形式靶向递送促耐受因子和胰岛素抗原，靶向关键的免疫细胞类型——树突状细胞（DC）。树突状细胞是吞噬性抗原呈递细胞，对于维持和启动对外来抗原的免疫性和对自身抗原的耐受性至关重要。这使得 DC 成为以颗粒形式提供的靶向递送药物的理想接受者。此外，用某些免疫调节剂对 DC 进行外源调节已被证明可以诱导 DC 的促耐受表型并改善 T1D。至关重要的是，使用 DC 靶向微粒 (MP) 进行疫苗接种有望纠正 T1D 自身免疫反应，而无需基于 DC 的细胞治疗所需的昂贵的离体操作。这使得具有广泛使用的潜力。 微米级的可生物降解聚合物颗粒具有可吞噬性，与非吞噬细胞相比，可有效促进封装因子向 DC 细胞内位点的递送。这些可吞噬颗粒可以通过靶向 DC 受体的表面固定分子进一步靶向 DC。较大（但仍足够小以可注射）、不可吞噬的可生物降解聚合物颗粒可将封装的因子控制释放到皮下注射部位的局部细胞外环境中。这些大颗粒中封装的因子由 DC 具有同源细胞表面受体的生物活性因子组成。该提案的目的是设计一种皮下注射的双 MP 疫苗系统，包括 i.) 可吞噬 DC 靶向 MP，将抗原和免疫调节因子（胰岛素和维生素 D3）传递到细胞内位点； ii.) 不可吞噬的 MP，用于递送细胞外因子（GM-CSF 和 TGF-b1），用于 DC 募集和耐受诱导。我们期望在非肥胖糖尿病 (NOD) 小鼠中产生促耐受性 DC 表型并促进调节性 T 细胞的诱导、自身反应性 T 细胞的抑制以及预防和逆转糖尿病。我们假设，双 MP 系统中多种成分的组合将比单成分制剂（无论是 MP 或可溶形式）更有效地提供强大、持久的抗原特异性免疫抑制。目标 1 是制定双 MP 系统，通过表征 DC 表型（激活、未成熟或耐受性）和 T 细胞反应（刺激、Th1、Th2、Treg 或 Th17）进行体外测试。目标 2 是评估双 MP 制剂在体内的能力，旨在预防和逆转 NOD 小鼠的糖尿病。这种新颖且创新的方法有望纠正 T1D 自身免疫反应，并代表了一种简单的、可临床转化的系统。