K-Channels in Lymphocyte Function and Autoimmunity

对抗原特异性免疫反应的功能性阻断或消除 在不影响全身免疫功能的情况下，必须将其视为寻求治疗的圣杯 自身免疫性疾病。实现这一崇高目标的一种方法是压制和/或消除 效应记忆(TEM)细胞与许多自身免疫的发病机制有关 在不影响其他免疫细胞的情况下预防疾病。本提案的重点是电压门控Kv1.3 瞬变细胞钾通道作为自身免疫性疾病的治疗靶点。与疾病相关 多发性硬化(MS)、1型糖尿病(T1 DM)患者自身反应性T细胞的变化 类风湿关节炎(RA)是一种Kv1.3通道表达升高的透射电子显微镜细胞。我们有 开发了高度特异的Kv1.3抑制剂，这些药物选择性地抑制钙信号转导， 细胞因子的产生、增殖和体内迁移不影响其他T细胞 子集。在概念验证研究中，Kv1.3阻滞剂预防和/或治疗大鼠模型中的疾病 MS、T1 DM、RA、接触性皮炎、迟发型超敏反应(DTH)和骨吸收 与牙周炎有关。这些阻滞剂在动物模型中具有极好的安全性。他们确实是这样 不影响对病毒和细菌病原体的急性保护性免疫反应。特定的 Kv1.3阻滞剂为静音自身反应提供了一种令人兴奋的新治疗方法 而不会影响保护性免疫反应。这项提案中概述的研究将 我们的主要候选药物ShK-186在多发性硬化症患者中的临床试验基础。目标1将 明确ShK-186-S治疗慢性复发缓解期的机制 DA大鼠实验性自身免疫性脑脊髓炎(CR-EAE)--人类MS模型 验证ShK-186通过一种方法消除致病的中枢神经系统自身抗原特异的横纹组织细胞的假设 我们称之为忽视致死的机制，而其他免疫细胞，特别是抑制疾病的细胞 调节性T细胞的增殖没有减弱，因为它们受到ShK-186抗性KCa3.1的保护 频道。目标2将描述ShK-186治疗的治疗剂量、频率和持续时间 以更好地预测人体研究。在目标3中，我们将使用膜片钳记录和流动 流式细胞术检测Kv1.3高表达是否是多发性硬化的可靠生物标志物 将有助于追踪Kv1.3阻滞剂治疗。目标3的第二个目标是研究渠道 食蟹猴T细胞的表达，因为该物种被广泛用于毒理学研究 对人类免疫调节疗法的评价，是我们启用IND的非人类灵长类动物的选择 毒物学研究。我们将确定食蟹猴T细胞的K通道表型 我们将评价ShK-186和S对猴横纹夜蛾细胞增殖的抑制作用。

Functional blockade or elimination of antigen-specific immune responses without impacting general immune function must be considered a holy grail in the quest to treat autoimmune disease. One approach to achieving this lofty goal is to suppress and/or eliminate effector memory (TEM) cells that have been implicated in the pathogenesis of many autoimmune diseases without affecting other immune cells. This proposal focuses on the voltage-gated Kv1.3 potassium channel in TEM cells as a therapeutic target for autoimmune diseases. Disease-associated autoreactive T cells in patients with multiple sclerosis (MS), type-1 diabetes mellitus (T1DM) and rheumatoid arthritis (RA) are TEM cells with elevated expression of Kv1.3 channels. We have developed highly specific inhibitors of Kv1.3 and these selectively suppress calcium signaling, cytokine production, proliferation, and in vivo migration of TEM cells without affecting other T cell subsets. In proof-of-concept studies, Kv1.3 blockers prevent and/or treat disease in rat models of MS, T1DM, RA, contact dermatitis, delayed type hypersensitivity (DTH) and bone resorption associated with periodontitis. These blockers have excellent safety profiles in animal models. They do not compromise the acute protective immune response to viral and bacterial pathogens. Specific Kv1.3 blockers, provide an exciting new therapeutic approach to mute autoreactive responses without compromising the protective immune response. The studies outlined in this proposal will lay the groundwork for clinical trials in patients with MS with ShK-186, our lead candidate. Aim 1 will define the mechanisms underlying ShK-186¿s therapeutic effect in chronic relapsing-remitting experimental autoimmune encephalomyelitis (CR-EAE) in DA rats, a model for human MS. We will test the hypothesis that ShK-186 eliminates disease-causing CNS autoantigen-specific TEM cells by a mechanism we term ¿death by neglect¿ while other immune cells, particularly disease-suppressing regulatory T cells, proliferate unabated because they are protected by the ShK-186-resistant KCa3.1 channel. Aim 2 will characterize the therapeutic dose, frequency and duration of ShK-186 treatment in CR-EAE to better predict human studies. In Aim 3 we will use patch-clamp recording and flow cytometry to determine whether Kv1.3high expression is a reliable biomarker for MS, and whether it will be useful for tracking Kv1.3 blocker therapy. A second goal of aim 3 is to study channel expression in T cells from Cynomolgus monkeys because this species is widely used for toxicological evaluation of human immune modulating therapies and is our non-human primate choice for IND-enabling toxicological studies. We will define the K-channel phenotype of cynomolgus monkey T cell subsets, and we will evaluate ShK-186¿s effectiveness in suppressing monkey TEM cell-proliferation.