BLR&D Research Career Scientist Award Application

我的神经免疫学研究项目的使命是发展对 影响中枢神经系统的自身免疫、脱髓鞘和神经变性过程 并确定和测试可推向市场以治疗和/或治愈的与疾病相关的新疗法 这些条件。退伍军人目前患上了难治性慢性神经系统疾病，如多发性 硬化症(MS)和中风，与服务相关的伤害，包括创伤性脑损伤(TBI)和创伤后 应激障碍(PTSD)，以及酒精和甲基苯丙胺的药物滥用。由本署进行的研究 实验室与这些破坏性疾病高度相关，因为我们开发了一种新的治疗方法 靶向共同的潜在机制，即促进中枢神经系统慢性炎症的MIF/CD74轴 和其他纸巾。巨噬细胞移动抑制因子及其同系物D-DT 是一种高度炎性的细胞因子，在结合时会触发其他炎症因子的释放 并通过它们共同的受体CD74发出信号，CD74是一种将自体和外源多肽加载到 抗原提呈细胞(APC)上的MHC II类分子。MIF/CD74信号转导的结果是多肽特异性的 效应细胞的激活和炎性细胞从血液到中枢神经系统的募集。我们最初的独特 治疗性构建物，称为RTL1000，由α-β(MS)的DR DR2 1和DR DR2 1结构域连接而成 风险因素)与髓鞘少突胶质细胞糖蛋白(MOG)35-55共价连锁。此构造具有 与天然存在的MHC II类/多肽T细胞受体配体构象相似，但可诱导T细胞 当APC上没有细胞结合的共刺激分子时，耐受性以可溶的形式存在。这一构造 在实验性自身免疫的小鼠模型中具有免疫调节和神经保护作用 脑脊髓炎，EAE)，在RTL1000女士的一项第一阶段研究中被证明是安全和耐受性良好的 很快将在多剂量多发性硬化症临床试验中进行测试。在理解药物的潜在影响方面的重大突破 RTL1000发生于2013年，发现CD74是RTL1000的细胞受体。这导致了 统一发现RTL1000可以竞争性地抑制MIF和D-DT与CD74的结合，从而 短路MIF/CD74信号，基本上存在于所有VA靶向中枢神经系统的情况下。分子 MIF结合模型揭示了两个离散的CD74区域，它们与MIF和MIF上的同源序列结合 D-DT和RTL1000的DRα1部分，从而解释了RTL1000的S竞争抑制。然而， RTL1000仅可用于约60%表达HLA-DR2的多发性硬化症患者。因此，我们设计了一种新的 构建了DRα1-MOG-35-55，它保留了RTL1000的活性，并能调节CD74和 竞争性阻断MIF结合，对慢性EAE有显著的治疗效果和神经保护作用。 值得注意的是，RTL1000和DRα1-MOG-35-55在治疗雄性小鼠慢性EAE方面更有效，这是因为 雌激素受体(ESR1)对女性的拮抗作用。A~600对MIF、D-DT和CD74的评价 主题临床研究(将在PNAS上发表)显示，男性受试者具有高表达基因 对于MIF(和D-DT)来说，发生进展性MS的风险增加，这一独特的观察结果提高了 早期使用RTL1000或DRα1-MOG-35-55治疗男性可能会阻止转化为 进展性多发性硬化症，对于患有进展性疾病的多发性硬化症受试者可能是有效的。FDA工业 DRα1-MOG-35-55临床前研究的批准将允许所有MS患者接受治疗，而无论 由于其非多态的、普遍的表达，所有人都能容忍，因此 可以在没有组织分型的情况下注射。更重要的是，我们已经证明了RTL1000和 Drα1-mog-35-55构建物还可以在实验小鼠模型中治疗其他中枢神经系统疾病，包括 实验性卒中、血管性痴呆、创伤性脑损伤和甲基苯丙胺诱导的认知 残疾，从而为患有这些破坏性疾病的退伍军人提供了一种新的潜在治疗方法。

The mission of my Neuroimmunology Research Program is to develop a deep biological understanding of autoimmune, demyelinating and neurodegenerative processes that affect the central nervous system (CNS) and to identify and test novel disease-relevant therapies that can be brought to market to treat and/or cure these conditions. Veterans are currently developing intractable chronic neurological diseases such as multiple sclerosis (MS) and stroke, service related injuries including traumatic brain injury (TBI) and post-traumatic stress disorder (PTSD), and substance abuse of alcohol and methamphetamine. Studies carried out by our laboratory are highly relevant to these devastating conditions due to our development of a novel therapy that targets a common underlying mechanism, the MIF/CD74 axis that promotes chronic inflammation in the CNS and other tissues. MIF (macrophage migration inhibitory factor) and its homolog D-DT (D-dopachrome tautomerase) are highly inflammatory cytokines that trigger release of other inflammatory factors upon binding and signaling through their common receptor, CD74, a chaperone for loading self and foreign peptides into MHC class II molecules on antigen presenting cells (APC). The result of MIF/CD74 signaling is peptide-specific Teffector cell activation and recruitment of inflammatory cells from blood into the CNS. Our initial unique therapeutic construct, called RTL1000 is comprised of linked DRα1 and DRβ1 domains of HLA-DR2 (an MS risk factor) covalently linked to myelin oligodendrocyte glycoprotein (MOG) 35-55. This construct has conformational similarity to naturally occurring MHC class II/peptide T cell receptor ligands, but induces T cell tolerance when present in soluble form without cell-bound co-stimulatory molecules on APC. This construct has immunoregulatory and neuroprotective properties in a mouse model of MS (experimental autoimmune encephalomyelitis, EAE) and was shown to be safe and well tolerated in a Phase 1 study in MS. RTL1000 will soon be tested in a multi-dose MS clinical trial. The major breakthrough in understanding the potent effects of RTL1000 occurred in 2013 with the discovery of CD74 as the cellular receptor for RTL1000. This led to the unifying discovery that RTL1000 could competitively inhibit binding of both MIF and D-DT to CD74 and thus short-circuit MIF/CD74 signaling that is present in essentially all of the VA targeted CNS conditions. Molecular modeling of MIF binding revealed two discrete CD74 regions that bound to homologous sequences on MIF and D-DT and to the DRα1 moiety of RTL1000, thus explaining RTL1000’s competitive inhibition. However, RTL1000 can only be used in ~60% of MS subjects that express HLA-DR2. We thus designed a new construct, DRα1-MOG-35-55 that retained the activities of RTL1000 and could modulate CD74 and competitively block MIF binding, resulting in a significant treatment effect and neuroprotection in chronic EAE. Of interest, RTL1000 and DRα1-MOG-35-55 were more effective in treating chronic EAE in male mice due to an antagonist effect of estrogen receptor (ESR1) in females. An evaluation of MIF, D-DT and CD74 in a ~600 subject clinical study (to be published in PNAS) revealed that male subjects with a high expression genotype for MIF (and D-DT) had an increased risk of developing progressive MS. This unique observation raises the possibility that early treatment of males with RTL1000 or DRα1-MOG-35-55 might prevent conversion to progressive MS and potentially would be effective for treating MS subjects with progressive disease. FDA IND approval of DRα1-MOG-35-55 preclinical studies would allow treatment of all MS subjects regardless of the HLA type due to its non-polymorphic, universal expression that would be tolerated by all humans and thus could be injected without tissue typing. Of broader importance, we have demonstrated that RTL1000 and DRα1-MOG-35-55 constructs can also treat other CNS conditions in experimental mouse models including experimental stroke, vascular dementia, traumatic brain injury and methamphetamine induced cognitive disability, thus providing a novel potential therapy for Veterans that develop these devastating conditions.