A Targeted Approach to Managing Salivary Gland Inflammation Using Resolvins

摘要 干燥综合征(SS)是一种以慢性炎症为特征的自身免疫性疾病 唾液腺的分泌功能。诊断特征包括口干，淋巴细胞渗透 并在血浆中发现抗核自身抗体。SS极大地降低了患者的生活质量 生命，尽管已经做了广泛的调查来了解它，但原因和有效的治疗 因为这种疾病仍然是未知的。鉴于目前治疗的高度需要和局限性， 开发新的治疗方法以减少炎症和恢复SG的分泌功能是至关重要的。我们的 以前的研究表明，SG表达AT-RvD1受体ALX/FPR2来阻断促炎作用 细胞因子信号转导，从而促进SS的细胞存活和组织完整性。最近，我们展示了 AT-RvD1对类SS NOD/ShiLtd J小鼠的治疗完全保留了分泌功能，下调了Pro-RvD-1的表达 炎性细胞因子的表达，并促进信号通路的分解。令人鼓舞的是，这些 然而，结果可能是，在AT-RvD1处理的SS小鼠中，淋巴细胞浸润持续存在，增加了 未来促炎细胞因子的产生和功能减退的可能复发。此外，我们面临着 临床应用必须克服的与解析剂本身的性质有关的问题 已实现。具体地说，很大一部分给药的解毒剂(包括Rvd1)没有达到他们的 预期的目的地具有所需的规律性，因为它们会被二十烷类化合物迅速灭活 氧化还原酶(EOR)及其作用可广泛分布于全身。此外， 不稳定的问题延伸到AT形式，它们在某种程度上不太容易失活。此外 对于这些持久性问题，解决方案相对昂贵，当这些问题结合在一起时，会引发 关于任何需要将大剂量引入血液的治疗的可行性的问题。在……里面 我们认为，所有这些问题都可以通过将AT-Rvd1逆行注入唾液来解决 管道，这可以允许AT-RvD1所展示的好处(即，保存和/或恢复唾液 分泌物)将被保留，同时解决前面讨论的薄弱领域。此外，为了更好地利用 利用AT-RvD1，我们将使用敲除小鼠来研究它激活ALX/FPR2的机制 该受体(ALX/FPR2-/-)。接下来，我们将描述ALX/FPR2信号机制在小鼠和 人类细胞。最后，我们将确定ALX/FPR2信号通路是否在小SG中发生改变 活组织检查(包括有无SS)，从而进一步将我们的发现扩展到人类疾病。我们的整体 假设通过局部AT-RvD1治疗，健康的SG功能将会恢复。目标1将演示 就地交付AT-Rvd1以恢复SG功能的好处。AIM 2将确定ALX/FPR2信令 机制和AIM 3将ALX/FPR2信号机制翻译成人类。

ABSTRACT Sjögren's syndrome (SS) is an autoimmune disease characterized by chronic inflammation and diminished secretory function of the salivary glands (SG). Diagnostic features include dry mouth, lymphocytic infiltration into the SG and presence of antinuclear autoantibodies in plasma. SS greatly decreases the patient’s quality of life, and although extensive investigation has been done to understand it, causes of and effective treatments for the disease are still unknown. In light of the high degree of need and the limitations of current therapies, development of novel treatments to decrease inflammation and restore SG secretory function is essential. Our previous studies demonstrated that SG express the AT-RvD1 receptor ALX/FPR2 to block pro-inflammatory cytokine signaling, thereby promoting cell survival and tissue integrity in SS. More recently, we demonstrated that AT-RvD1 treatment in SS-like NOD/ShiLtJ mice fully preserves secretory functioning, downregulates pro- inflammatory cytokine expression and promotes pro-resolving signaling pathways. Encouraging as these results may be, however, lymphocytic infiltration persists in AT-RvD1 treated SS mice, raising the possibility of future pro-inflammatory cytokine production and possible recurrence of hypofunction. Moreover, we face issues related to the properties of resolvins themselves that must be overcome for clinical applications to be achieved. Specifically, a significant portion of administered resolvins (RvD1 included) do not reach their intended destination with the desired regularity because they are quickly inactivated by eicosanoid oxidoreductases (EOR) and their effects can be diffusely distributed throughout the body. Furthermore, the problem of instability extends to the AT forms, which are somewhat less susceptible to inactivation. In addition to these issues of durability, resolvins are relatively expensive, and when taken together, these concerns raise questions about the feasibility of any treatment requiring large doses to be introduced into the blood stream. In response, we believe that all of these issues may be addressed by infusing AT-RvD1 retrograde to the salivary ducts, which could allow the benefits demonstrated by AT-RvD1 (i.e., preserved and/or restored saliva secretion) to be retained while addressing areas of weakness discussed earlier. Moreover, to better exploit the use of AT-RvD1, we will study the mechanisms by which it activates the ALX/FPR2 using a knock-out mouse for this receptor (ALX/FPR2-/-). Next, we will describe the ALX/FPR2 signaling mechanisms in mouse and human cells. Finally, we will determine whether ALX/FPR2 signaling pathways are altered in minor SG biopsies (both with and without SS), thereby further extending our findings to the human disease. Our overall hypothesis is that healthy SG functioning will be restored by local AT-RvD1 treatment. Aim 1 will demonstrate the benefits of delivering AT-RvD1 locally for restoring SG function. Aim 2 will determine ALX/FPR2 signaling mechanisms and Aim 3 will translate ALX/FPR2 signaling mechanisms to humans.