Tunable Assembly of Regulatory Immune Signals to Promote Myelin-specific Tolerance

在自身免疫性疾病期间，身体识别并攻击“自我”分子。目前的治疗方法是不能治愈的 并要求终身遵守。此外，现有的治疗方法--虽然有益，但广泛有效，可以离开 患者的免疫功能受损。这些挑战激发了人们对控制自身免疫的极大兴趣 疫苗样的特异性，以保持正常的免疫功能。临床前和临床研究都在测试这一点 多发性硬化症(MS)的想法，一种影响退伍军人的比例失调的疾病。多发性硬化是一种神经退行性疾病 由中枢神经系统(CNS)髓鞘错误发作引起的疾病。因此，一种实验性的疗法 IDEA涉及共同给药髓磷脂多肽和耐受信号以促进髓鞘特异性调节T细胞 控制多发性硬化症的细胞(Treg)在最近的一些试验中，一些6-8个髓鞘表位的小集合正在作为一条路线进行测试 以跨越许多-尽管不是所有-患者的髓鞘反应范围。有趣的是，有一组新的路径- 与髓鞘驱动的炎症相关的是Toll样受体(TLRs)。在健康人中，TLRs检测到 病原体相关模式，以调动先天免疫。然而，新的研究表明，TLR信号--例如 TLR9-在MS患者和MS模型(如EAE、RR-EAE)中升高。在后者中，抑制TLR9函数 减少炎症，同时也促进Treg和改善疾病。淋巴结(LNS)和脾 控制髓鞘反应性T细胞向炎性T细胞极化的关键组织(如TH17)或 特雷格。因此，当髓鞘出现在LNS中时，引导T细胞分化的策略-例如，联合传递 可能产生大量的髓鞘特异性Treg，从而阻止致病免疫细胞 没有广泛的镇压。纳米技术为这一目标提供了独特的能力，包括联合传递自我 抗原和调节信号、靶向和可调释放。然而，许多聚合物颗粒和其他 生物材料表现出触发炎症信号的内在特征，炎症信号可能会加剧自身免疫。 模仿生物材料的吸引人的特征，同时消除炎性“载体”效应的策略可能是 针对多发性硬化症的新疗法的变革性应用这一更新的VA优点应用程序使用多离子免疫信号 先进的新型纳米结构胶囊完全由调节免疫线索和髓鞘抗原构建。这些 免疫聚电解质多层膜(IPEM)通过静电作用组装在模板上， 它被移除，留下胶囊，将髓鞘与针对TLR9(GPG)的调节TLR配体并列。 由于没有载体，iPEMS中的信号密度相对于聚合物或脂类包埋而言非常高 货物(例如纳米颗粒)。在最初的颁奖过程中，这种自我抗原和调节的高密度并列 CUE(GPG)被证明促进髓鞘特异性T细胞向Treg分化，而不是 炎性T细胞。同样，对多发性硬化症患者样本的初步研究表明，髓鞘/GPG iPEM抑制髓鞘- 引发炎症。在MS的复发-缓解(RR-EAE)和进行性(EAE)模型中，iPEM反转 疾病导致的瘫痪。在持续的退伍军人管理局支持下，该项目将生成剩余的临床前数据 需要激励未来临床开发一种疗法，为退伍军人及其家人提供安全的 和有效的疫苗样免疫疗法治疗多发性硬化症广泛的目标是确定TLR信号在T细胞中的作用 极化和有效性，并表明这些影响在疾病模型和一系列 现有的、身份不明的男性和女性退伍军人多发性硬化症患者样本。我们的计划得到了一个多学科的支持 由生物工程师、免疫学家和多发性硬化症临床医生组成的团队，在这方面和其他方面都有有效合作的历史 项目的重点是抗原特异性耐受性。具体目标是1)确认iPEM改变TLR信令和 APC的功能和T细胞极化的链接变化，2)定义IPEM驱动的结构和功能变化 3)确定疗效的持久性和中枢神经系统的病理变化 在EAE/RR-EAE期间，4)Show iPEM调节多发性硬化症患者样本中的TLR信号和抗原特异性反应。

During autoimmune disease, the body identifies and attacks “self” molecules. Current therapies are not curative and require life-long compliance. Further, existing therapies – while beneficial, are broadly acting and can leave patients immunocompromised. These challenges have sparked great interest in controlling autoimmunity with vaccine-like specificity to preserve normal immune function. Both pre-clinical and clinical studies are testing this idea in multiple sclerosis (MS), a disease that disproportionality impacts Veterans. MS is a neurodegenerative disease driven by mistaken attack of myelin in the central nervous system (CNS). Thus, an experimental therapy idea involves co-administration of myelin peptide and tolerizing cues to promote myelin-specific regulatory T cells (TREG) that control MS. In some recent trials, small sets of 6-8 myelin epitopes are being tested as a route to span the range of myelin reactivity in many - though not all - patients. Interestingly, one set of pathways newly- associated with myelin-driven inflammation are toll-like receptors (TLRs). In healthy people, TLRs detect pathogen-associated patterns to mobilize innate immunity. However, new work shows TLR signaling - such as TLR9 - is elevated in MS patients and MS models (e.g., EAE, RR-EAE). In the latter, suppressing TLR9 function reduces inflammation, while also promoting TREG and improving disease. Lymph nodes (LNs) and spleen are key tissues that control polarization of myelin-reactive T cells toward either inflammatory T cells (e.g., TH17) or TREG. Thus, strategies that guide T cell differentiating when myelin is presented in LNs – for example, co-delivery of regulatory cues – could generate large populations of myelin-specific TREG that stop pathogenic immune cells without broad suppression. Nanotechnology offers unique capabilities for this goal, including co-delivery of self- antigen and regulatory cues, targeting, and tunable release. However, many polymer particles and other biomaterials exhibit intrinsic features that trigger inflammatory signaling, which could exacerbate autoimmunity. Strategies that mimic attractive features of biomaterials, while eliminating inflammatory “carrier” effects could be transformative for new therapies for MS. This Renewal VA Merit application uses polyionic immune signals to advance novel nanostructured capsules built entirely from regulatory immune cues and myelin antigen. These immune polyelectrolyte multilayers (“iPEMs”) are assembled through electrostatic interactions on a template, which is removed to leave capsules that juxtapose myelin with a regulatory TLR ligand against TLR9 (GpG). Since there is no carrier, the density of signals in iPEMs is very high relative to polymer or lipids encapsulating cargo (e.g., nanoparticles). During the initial award, this high density juxtaposition of self-antigen and regulatory cue (GpG) was shown to promote differentiation of myelin-specific T cells toward TREG and away from inflammatory T cells. Likewise, pilot studies with MS patient samples reveal myelin/GpG iPEMs restrain myelin- driven inflammation. In relapsing-remitting (RR-EAE) and progressive (EAE) models of MS, iPEMs reverse disease-driven paralysis. With continued VA support, this project will generate the remaining pre-clinical data needed to motivate future clinical development of a therapy that could offer Veterans and their families a safe and effective vaccine-like immunotherapy for MS. The broad goal is to define the role of TLR signaling in T cell polarization and efficacy, and show these effects are durable and robust across disease models and a set of existing, de-identified male and female Veteran MS patient samples. Our plan is supported by a multidisciplinary team of bioengineers, immunologists, and MS clinicians with a history of effective collaboration on this and other projects focused on antigen-specific tolerance. The specific aims are 1) Confirm iPEMs alter TLR signaling & function in APCs and link changes to T cell polarization, 2) Define iPEM-driven structural and functional changes associated with tolerance in LNs & spleen, 3) Ascertain the durability of efficacy and changes in CNS pathology during EAE/RR-EAE, 4) Show iPEMs regulate TLR signaling & antigen-specific response in MS patient samples.