RESOLUTION OF CYTOKINE-MEDIATED SALIVARY GLAND INFLAMMATION

摘要 干燥综合征(SS)是一种以分泌减少为特征的慢性炎症性自身免疫病 外分泌腺的功能。治疗性腺功能减退的方法仅限于使用唾液替代品和 只能提供暂时缓解的药物。鉴于当前的高度需要和局限性 在治疗方面，开发替代疗法以恢复功能是至关重要的。解毒剂(RV)，它们是 它们是高度有效的脂质介体，为更好地治疗炎症性疾病提供了可行的替代方案 作为党卫军。我们上一个资助期的结果显示如下：a)唾液腺表达 RvD_1和AT-RvD_1受体(ALX/FPR2)；b)激活ALX/FPR2阻断促炎细胞因子 信号，从而促进唾液细胞的细胞存活和组织完整性；c)Rvd1生物合成途径 在唾液上皮中有功能；d)ALX/FPR2丢失导致未消退的炎症和SMG 体内功能障碍；e)AT-RvD1治疗SS样NOD/ShiLtd J小鼠完全保留了分泌功能， 下调促炎症细胞因子的表达，促进促分解信号通路；f) 联合应用AT-RvD_1和地塞米松(DEX)减少小鼠唾液腺淋巴细胞浸润 与单独使用AT-RvD1进行比较，优化了人SMG细胞的三维培养体系 为了研究ALX/FPR2信号和h)，证实了人唾液中表达分解素脂 调解人。然而，尽管这些发现可能是有希望的，但我们面临着必须 为解析剂的临床应用所需攻克。具体地说，解决方案(包括RvD1)不 有规律地到达预定的目的地，因为它们很快就会被二十烷类化合物灭活 氧化还原酶或Eors进入人体后，血浆半衰期较短(约3小时) 具有遍及全身的影响(因此它们不能专门针对发炎的区域，如 将需要对炎症性疾病进行最佳治疗)。此外，不稳定的问题 扩展到AT形式，它们不太容易失活，但仍然表现出低于 理想的稳定性。然而，我们认为所有这些问题都可以通过使用弹性蛋白样多肽来解决。 纳米粒子(ITEP NPs)，这是一种生物相容的胶束状分子，经过改造含有AT-RvD1。 具体地说，ITEP NPs能够保护AT-RvD1免于失活，延长血浆半衰期和 容易渗入发炎的血管，从而使它们聚集在发炎的区域。这个 这项建议的主要目的是确定AT-RvD1治疗是否可以减轻SS症状。目标 1：将在NOD/ShiLtd J SS小鼠模型中展示AT-RvD1治疗的有效性。目标2：将翻译 AT-RvD1对人唾液细胞的治疗和目标3：将通过交付加强AT-RvD1的治疗 使用ITEP NP的系统。

ABSTRACT Sjögren's syndrome (SS) is a chronic inflammatory autoimmune disease characterized by diminished secretory function of the exocrine glands. Treatments for hyposalivation are limited to use of saliva substitutes and medications that provide only temporary relief. In light of the high degree of need and the limitations of current therapies, development of alternative treatments to restore functioning is essential. Resolvins (Rv), which are which are highly potent lipid mediators, offer a viable alternative for better treating inflammatory diseases such as SS. Results from our previous funding period demonstrated the following: a) salivary glands express the RvD1 and AT-RvD1 receptor (ALX/FPR2); b) activation of ALX/FPR2 blocks pro-inflammatory cytokine signaling, thereby promoting cell survival and tissue integrity in salivary cells; c) RvD1 biosynthetic pathways are functional in salivary epithelium; d) loss of ALX/FPR2 results in unresolved inflammation and SMG dysfunction in vivo; e) AT-RvD1 treatment in SS-like NOD/ShiLtJ mice fully preserves secretory functioning, downregulates pro-inflammatory cytokine expression and promotes pro-resolving signaling pathways; f) treatment combining AT-RvD1 and dexamethasone (DEX) reduces salivary gland lymphocytic infiltration in comparison to AT-RvD1 alone, g) a three-dimensional culture system using human SMG cells was optimized for the study of ALX/FPR2 signaling and h) human saliva was demonstrated to express resolvin lipid mediators. Promising though these findings may be, however, we face significant issues that must be overcome for the clinical application of resolvins to be achieved. Specifically, resolvins (RvD1 included) do not reach their intended destination with desired regularity because they are quickly inactivated by eicosanoid oxidoreductases or EORs upon introduction to the body, have a short plasma half-life (approximately 3 h) and have effects that are spread throughout the body (such that they cannot specifically target inflamed areas, as would be needed for optimal treatment of inflammatory diseases). Furthermore, the problem of instability extends to the AT forms, which are somewhat less susceptible to inactivation but still demonstrate less than ideal stability. However, we believe that all of these issues may be addressed by use of elastin-like polypeptide nanoparticles (iTEP NPs), which are biocompatible micelle-like molecules engineered to contain AT-RvD1. Specifically, iTEP NPs are capable of protecting AT-RvD1 from inactivation, increasing plasma half-life and easily permeating inflamed blood vessels, thereby allowing them to congregate in areas of inflammation. The main goal of this proposal is to determine whether SS symptoms can be reduced by AT-RvD1 treatment. Aim 1: will demonstrate the utility of AT-RvD1 treatment in the NOD/ShiLtJ SS mouse model. Aim 2: will translate AT-RvD1 treatment to human salivary cells and Aim 3: will enhance AT-RvD1 treatment through a delivery system using iTEP NPs.