Methods to modulate GI inflammatory and infectious diseases

传染病是重大的医疗问题。因为这种疾病的发病机制是由 细菌和病毒，常用的药物都是小分子药物。然而，细菌和病毒经常 突变，使开发护理药物的标准具有挑战性。因此，疫苗，这是 注射活的或减毒的抗原以产生对病原体的免疫力的免疫疗法有 用于预防和治疗。然而，疫苗对胃肠道的效果较差。 (GI)引起粘膜免疫的传染病需要广泛激活GI免疫细胞。在这 研究，我们的目标是开发一种疫苗佐剂系统，可以通过口服途径，并选择性地 激活胃肠道免疫细胞以诱导强大的粘膜免疫，这将通过以下特定的目的来追求。 目的：1.我们将识别一种新的小分子，它可以在没有GI免疫细胞的情况下 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检测方法 (HTS)。使用HTS测试，我们将发现一种可以诱导促炎的命中化合物 原代树突状细胞分泌的细胞因子TNF-α，但不能诱导胃肠道内皮细胞分泌肿瘤坏死因子-α 卡科-2。然后，我们将开发一种适合口服的纳米药物载体。我们将发展 可保护HIT化合物在胃肠道中不被降解的聚合物纳米颗粒，以及 增强肠道免疫细胞的摄取，所有这些都是诱导肠道粘膜免疫的重要因素 胃肠道。 目的2.HIT化合物包裹纳米粒(简称纳米佐剂)的体内检测 在啮齿动物模型中的安全性和有效性。纳米粒的生物分布和药代动力学 使用IVIS荧光成像和LC/MS分析测量器官的组织学分析 目的：研究纳米佐剂的潜在毒性。接下来，我们将研究免疫刺激作用。 检测T细胞和B细胞(小肠细胞、肠系膜淋巴)活化的纳米佐剂的疗效 结节)和粘膜免疫(肠道冲洗IgA、粪便IgA)。 在这项提案中，我们将集中精力检查口服纳米颗粒是否能传递小分子 通过HTS筛选和体内安全性和安全性研究，可在不引起胃肠道炎症的情况下诱导强大的粘膜免疫 疗效验证。该项目的长期目标是开发一种通用的口服免疫刺激佐剂 当与抗原源结合时，可用作疫苗，或作为补充免疫疗法 当与抗菌药物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.