Methods to modulate GI inflammatory and infectious diseases

传染病是重大的医疗保健问题。由于疾病的发病机制是由 细菌和病毒，常用的药物是小分子药物。然而，细菌和病毒通常 突变，并使其具有挑战性，以开发一个标准的护理药物。因此， 注射活的或减毒的抗原以产生针对病原体的免疫力的免疫治疗方案， 用于预防和治疗应用。然而，疫苗对胃肠道疾病的有效性较低， (GI)感染性疾病产生粘膜免疫需要广泛激活GI免疫细胞。在这 研究中，我们的目标是开发一种疫苗佐剂系统，可以通过口服途径， 激活胃肠道免疫细胞以引发强粘膜免疫，这将通过以下特定目标来实现。 目标1.我们将确定一种新的小分子，它可以特异性地激活GI免疫细胞， 引发慢性胃肠道炎症为此，我们将开发允许高通量筛选的体外测定法 (HTS)药物图书馆。使用HTS测定，我们将发现一种诱导促炎性细胞因子的命中化合物， 细胞因子，来自原代树突状细胞的TNF-α，但不诱导来自GI内皮细胞系的TNF-α Caco-2然后，我们将开发一种适合口服给药的纳米粒药物载体。我们将开发 聚合物纳米颗粒，其可以提供保护所述命中化合物免于在胃肠道中降解，以及 增强肠道中GI免疫细胞的摄取，所有这些对于诱导粘膜免疫都很重要， 胃肠道。 目标2.将在体内检查包封纳米颗粒的命中化合物（称为纳米佐剂） 在啮齿动物模型中的安全性和有效性。纳米颗粒的生物分布和药代动力学特征将被 将使用IVIS荧光成像和LC/MS分析以及器官的组织学分析进行测量。 研究纳米佐剂的潜在毒性。接下来，我们将研究免疫刺激 通过测量T细胞和B细胞活化（小肠细胞、肠系膜淋巴液）来测定纳米佐剂的功效 淋巴结）和粘膜免疫（肠洗液伊加、粪便伊加）。 在这项建议中，我们将集中精力研究口服纳米微粒递送小分子 通过HTS筛选和体内安全性，可引发强粘膜免疫而不诱导GI炎症， 功效验证。该项目的长期目标是开发一种通用的口服免疫刺激佐剂 当与抗原源组合时，其可用作疫苗，或用作补充免疫疗法， 当与抗生素或抗病毒药物联合治疗胃肠道感染性疾病时，

Infectious diseases are significant healthcare problems. As pathogenesis of the disease are triggered by bacteria and virus, commonly used medications are small molecule drugs. However, bacteria and virus often mutate and makes it challenging to develop a standard of care medications. Hence, vaccines, which are immunotherapy regimens to inject live or attenuated antigens to develop immunity against the pathogens are used for prophylactic and therapeutic applications. However, vaccines are less effective against gastrointestinal (GI) infectious diseases as generating mucosal immunity requires extensive activation of GI immune cells. In this study, we aim to develop a vaccine adjuvant system which can be delivered by oral route, and selectively activate GI immune cells to elicit strong mucosal immunity which will be pursued by following specific aims. Aim1. We will identify a novel small molecule which can specifically activate GI immune cells without triggering chronic GI inflammation. To do so, we will develop in vitro assays which allow high throughput screen (HTS) of drug libraries. Using HTS assay, we will discover a hit compound that induces pro-inflammatory cytokine, TNF-alpha from primary dendritic cells but does not induce TNF-alpha from GI endothelial cell line Caco-2. Then, we will develop a nanoparticle drug delivery carrier suitable for oral delivery. We will develop polymeric nanoparticles that can provide protection of the hit compound from degradation in the GI tract, and enhance uptake by GI immune cells in the intestine, all of which are important for inducing mucosal immunity in GI tract. Aim 2. Hit compound encapsulating nanoparticles (refer to as Nano-Adjuvant) will be examined for in vivo safety and efficacy in rodent animal models. Biodistribution and pharmacokinetic profile of nanoparticles will be measured using IVIS fluorescence imaging and LC/MS analysis and histological analysis of organs will be performed to investigate the potential toxicity of Nano-adjuvant. Next, we will examine the immunostimulatory efficacy of Nano-adjuvant by measuring the T cell and B cell activation (small intestinal cells, mesenteric lymph nodes) using flow cytometry, and mucosal immunity (Intestinal wash IgA, fecal IgA). In this proposal, we will focus our efforts to examine whether oral nanoparticulate delivery of small molecule can elicit strong mucosal immunity without inducing GI inflammation by HTS screening and in vivo safety and efficacy validations. Long term goal of the project is to develop a universal oral immunostimulatory adjuvant which can be used as a vaccine when combined with antigen sources, or as a complementary immunotherapy agent when combined with anti-biotics or anti-virus drugs to treat GI infectious diseases.