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: 8442857
• 负责人: Benjamin George Keselowsky
• 金额: $ 30.65万
• 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/8442857
• 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）由于对产生胰岛素的β细胞的自我破坏性免疫反应而产生的胰岛素不足而产生。尽管已知许多因素可以促进T1D实验系统中有利的免疫细胞反应，但由于不受控制的旁观者细胞，组织和器官的剂量不受控制，这些药物的全身静脉输送通常会导致明显的有害脱靶效应。该项目的重点是靶向耐耐受因子的靶向体内递送和胰岛素抗原，以颗粒形式，针对关键的免疫细胞类型树突状细胞（DCS）。树突状细胞对于对放弃抗原的维持和启动至关重要，对自我抗原的耐受性是吞噬细胞的抗原呈递细胞。这使DCS成为以颗粒形式提供的代理的靶向递送的理想接收者。此外，已证明具有某些免疫调节剂的DC的外源调节可诱导耐受性DC表型以及改善T1D。使用DC靶向微粒（MP）疫苗接种有望纠正T1D自身免疫反应，而无需基于直流的细胞疗法所需的昂贵的离体操纵。这使得有可能广泛使用。 微米大小的可生物降解聚合物颗粒是可吞噬的，可有效促进封装的因子传递到非少细胞上DC的细胞内部位。这些吞噬细胞颗粒可以通过固定靶向直流受体的分子表面表面靶向DC。较大（但仍然足够小，可以注射），不可刺激性可生物降解的聚合物颗粒可控制地下注射部位的局部细胞外环境的封装因子。这些大颗粒中的封装因子由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由I。）吞噬可吞噬的DC靶向MPS的皮下注射的双MP疫苗系统，该MP递送抗原和免疫调节因子（胰岛素和维生素D3）到细胞内部位；和ii。）不可用的MPS用于传递细胞外，DC募集和耐受性诱导的细胞外因子（GM-CSF和TGF-B1）。我们期望实现耐耐受性的直流表型，并促进调节性T细胞的诱导，抑制自动反应性T细胞，并预防非肥胖糖尿病（NOD）小鼠中的糖尿病。我们假设双重MP系统中多个组件的组合将比以MPS或可溶性形式更有效地提供坚固，耐用的抗原特异性免疫抑制。 AIM 1是制定双MP系统，通过表征DC表型（激活，不成熟或耐受性）和T细胞响应（刺激，TH1，TH2，Treg或Th17）来在体外对其进行测试。 AIM 2是评估体内双MP公式的能力，旨在预防和反向NOD小鼠中的糖尿病。这种新颖的创新方法有望纠正T1D中的自身免疫反应，并代表一个简单的临床转换系统。