Immunoregulatory effect of microparticle delivered STING agonist in the control of experimental autoimmune encephalomyelitis (EAE) and multiple sclerosis (MS)

微粒递送的 STING 激动剂在控制实验性自身免疫性脑脊髓炎 (EAE) 和多发性硬化症 (MS) 中的免疫调节作用

• 批准号: 10392967
• 负责人: Silva Markovic-Plese
• 金额: $ 19.6万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-15 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10392967
• 关键词: Acetals; Acids; Agonist; Animal Model; Anti-Inflammatory Agents; Antigen-Presenting Cells; Antigens; B-Lymphocytes; Bone Marrow; CD4 Positive T Lymphocytes; Cell Differentiation process; Cells; Chronic; Clinical; Cyclic AMP; Cyclic GMP; Data; Dendritic Cells; Dextrans; Dinucleoside Phosphates; Disease; Disease Progression; Dose; Encapsulated; Equilibrium; Experimental Autoimmune Encephalomyelitis; FOXP3 gene; Human; IL2RA gene; ITGAX gene; Immune; Immune Tolerance; Immunomodulators; In Vitro; Inflammatory Infiltrate; Interferon-beta; Interleukin-10; MHC Class II Genes; Mediating; Monoclonal Antibodies; Multiple Sclerosis; Mus; Organ; Pathogenicity; Patients; Peptides; Periodicity; Peripheral; Phagocytes; Phagosomes; Phenotype; Polymers; Production; Reaction Time; Regulatory T-Lymphocyte; Relapsing-Remitting Multiple Sclerosis; Reporting; Role; Signal Transduction; Stimulator of Interferon Genes; Testing; Therapeutic; Therapeutic Effect; Toxic effect; Translating; antigen binding; base; biomaterial compatibility; controlled release; cytokine; experimental study; immunomodulatory therapies; immunoregulation; macrophage; monocyte; multiple sclerosis patient; novel; particle; pre-clinical; reconstitution; side effect; transcriptome

While multiple antiinflammatory therapies are effective in suppressing relapsing-remitting multiple sclerosis (RRMS) disease progression, they have significant side effects and toxicity. We are still seeking a disease specific treatment that would provide an antigen-specific immune tolerance reconstitution. We used microparticle (MP)-encapsulated STING agonist cGAMP to increase its delivery into phagocytic antigen presenting cells (APCs), particularly dendritic cells (DCs). A novel acid-sensitive polymer, acetalated dextran (Ace-DEX) allows regulated MP breakdown due to its pH sensitivity in intracellular acidic compartments. Our preliminary data have demonstrated that the intracellular delivery of MPs encapsulating 2’3’cyclic GMP-AMP dinucleotide (cGAMP) suppressed experimental autoimmune encephalomyelitis (EAE), an animal model of MS, when administered before and at the peak of clinical disease. The therapeutic effect of cGAMP MPs in RR and chronic EAE was mediated via STING-induced IL-27 and IL-10 production, since the therapeutic effect was abrogated in IL-27ra-/- and IL-10-/- mice, while partial disease suppression was maintained in Ifnar-/- mice. Our central hypothesis is that intracellular cGAMP MP delivery will induce IL-27 and IL-10-producing tolerogenic DCs that regulate the differentiation and expansion of IL-10-producing iTregs. We will characterize cGAMP MP tolerogenic DC phenotype and function, as well as suppressive capacity of the induced regulatory CD4+IL-10+ cells, in particular their expression of FoxP3 and type 1 iTreg (Tr1) markers. We propose that presentation of MHC class II DR2-anchored peptide mixtures (DR2-APMs) by cGAMP MP-induced tolerogenic DCs from DR2+ patients with relapsing remitting multiple sclerosis (RRMS) may reconstitute antigen-specific immune tolerance. In order to identify the mechanisms of cAMP MP-mediated disease suppression, we will pursue following specific aims: 1) Demonstrate that cGAMP MP treatment induces iTreg cell expansion in EAE. The proposed studies will determine to what extent cGAMP MP treatment of EAE induces the expansion of CD4+CD25+IL-27R+IL-10+ Tregs in the peripheral immune organs and the CNS inflammatory infiltrates. Studies using aCD25mAb will confirm the role of iTregs in the therapeutic effect. The role of FoxP3+Tregs will be examined using transient inducible FoxP3+Treg depletion and mice with selective FoxP3+Treg IL-27R depletion. 2) In human in-vitro studies, we will characterize cGAMP MP-induced tolerogenic DCs and their capacity to induce iTregs in RRMS patients. Specific experiments will determine the capacity of cGAMP MP-treated monocytes and DCs to induce iTreg expansion, determine the transcriptome of cGAMP MP IL-27- and IL-10- induced Tregs and the reconstitution of their suppressive capacity. Finally, co-administration of cGAMP MPs with DR2-APMs to DCs from RRMS patients will test their capacity to induce antigen-specific tolerogenic DCs and Tregs. The study will provide preclinical data required to translate this therapeutic approach into clinical use in patients with RRMS

虽然多种抗炎疗法可有效抑制复发的多发性硬化症 （RRMS）疾病进展，它们具有显着的副作用和毒性。我们仍在寻找疾病 可以提供抗原特异性免疫耐受性重建的特定治疗方法。我们使用了 微粒（MP）包含的刺激性激动剂CGAMP将其递送到吞噬抗原中 呈现细胞（APC），特别是树突状细胞（DC）。一种新型的酸敏感聚合物，乙酰化葡萄糖 （ACE-DEX）由于其在细胞内酸性室中的pH敏感性而允许MP分解。我们的 初步数据表明，封装2'3'Cycle GMP-AMP的MP的细胞内递送 二核苷酸（CGAMP）抑制了MS动物模型的实验性自身免疫性脑膜炎（EAE） 当在临床疾病的峰值之前和处施用时。 CGAMP MPS在RR和 慢性EAE是通过刺激诱导的IL-27和IL-10产生介导的，因为治疗效果为 在IL-27RA - / - 和IL-10 - / - 小鼠中进行了牵引，而在IFNAR - / - 小鼠中保持了部分疾病抑制。我们的 中心假设是细胞内CGAMP MP递送将诱导IL-27和IL-10产生的耐受性 调节IL-10产生ITREGS的分化和扩展的DC。我们将表征CGAMP MP 耐受性直流表型和功能，以及诱导的调节性CD4+IL-10+的抑制能力 细胞，特别是它们的Foxp3和1型ITREG（TR1）标记的表达。我们建议 MHC II类DR2锚定的肽混合物（DR2-APMS）通过CGAMP MP诱导的DR2+的耐受性DC 复发释放多发性硬化症（RRMS）的患者可能会重建抗原特异性免疫耐受性。 为了确定CAMP MP介导的疾病抑制的机制，我们将追求 具体目的：1）证明CGAMP MP处理可诱导EAE中的ITREG细胞扩展。这 拟议的研究将确定CGAMP MP治疗EAE的范围诱导的扩展 周围免疫力和中枢神经系统炎性浸润中的CD4+CD25+IL-27R+IL-10+Tregs。研究 使用ACD25MAB将确认ITREG在治疗效果中的作用。 Foxp3+Tregs的角色将是 使用瞬态诱导FOXP3+Treg耗竭和具有选择性FOXP3+Treg IL-27R耗竭的小鼠检查。 2）在人类内研究中，我们将表征CGAMP MP诱导的耐受性DC及其能力 在RRMS患者中诱导ITREGS。具体实验将确定CGAMP MP处理的能力 单核细胞和DC诱导ITREG扩展，确定CGAMP MP IL-27-和IL-10-的转录组 诱发Treg及其抑制能力的重构。最后，CGAMP MPS的共同管理 从RRMS患者到DC的DR2-APMS将测试其诱导抗原特异性耐受性DC的能力 和Tregs。该研究将提供将这种治疗方法转化为临床用途所需的临床前数据 RRMS患者