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

虽然多种抗炎疗法在抑制复发-缓解型多发性硬化症方面是有效的 (RRMS)疾病进展，它们有显著的副作用和毒性。我们仍在寻找一种疾病 将提供抗原特异性免疫耐受重建的特定治疗。我们用了 微球包裹刺激剂cGAMP增加其对吞噬细胞抗原的转运 提呈细胞(APC)，尤其是树突状细胞(DC)。一种新型酸敏聚合物--缩醛葡聚糖 (ACE-DEX)由于其在细胞内酸性隔间的pH敏感性，允许调节MP的分解。我们的 初步数据表明，包裹2‘3’环状GMP-AMP的MPS的细胞内递送 二核苷酸(CGAMP)抑制实验性自身免疫性脑脊髓炎(EAE)，MS的动物模型， 在临床疾病之前和高峰期给药。环丙沙星治疗慢性阻塞性肺疾病的疗效观察 慢性EAE是通过刺痛诱导的IL-27和IL-10的产生而介导的，因为治疗效果是 在IL-27ra-/-和IL-10-/-小鼠中被取消，而在Ifnar-/-小鼠中保持部分疾病抑制。我们的 核心假设是细胞内cGAMP MP传递将诱导产生IL-27和IL-10的耐受性 调节产生IL-10的iTregs的分化和扩张的DC。我们将描述cGAMP MP 致耐受DC的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。我们建议提出 CGAMP MP诱导DR2+产生的耐受性DC的MHC-II类DR2锚定多肽混合物(DR2-APM) 复发缓解型多发性硬化症(RRMS)患者可重建抗原特异性免疫耐受。 为了明确cAMP MP介导的疾病抑制机制，我们将进行以下研究 具体目的：1)证实cGAMP MP可诱导EAE中iTreg细胞的增殖。这个 拟议的研究将确定cGAMP MP治疗EAE在多大程度上诱导EAE的扩张 外周免疫器官中有CD4+CD25+IL-27R+IL-10+Tregs，中枢神经系统炎性细胞浸润。研究 使用CD25mAb将证实iTregs在治疗效果中的作用。FoxP3+Treg的作用将是 用瞬时诱导的FoxP3+Treg耗竭和选择性FoxP3+Treg IL-27R耗竭的小鼠进行检测。 2)在人类体外研究中，我们将表征cGAMP MP诱导的耐受性DC及其能力 目的：在RRMS患者中诱导iTregs。具体实验将确定cGAMP MP处理后的容量 单核细胞和DC诱导iTreg扩增，测定cGAMP MP IL-27-和IL-10-的转录组 诱导树突起及其抑制能力的重建。最后，cGAMP MPS的共同管理 RRMS患者DC与DR2-APM联合诱导抗原特异性耐受树突状细胞的能力 还有特雷格斯。这项研究将提供将这种治疗方法转化为临床应用所需的临床前数据。 在RRMS患者中