Engineering bacterially derived immunomodulants:a novel IBD therapeutic approach

生物工程细菌源性免疫调节剂：治疗IBD的新途径 项目摘要 长期目标是开发新的，有效的治疗方法，利用免疫调节特性， 用于治疗炎症性肠病（IBD）的细菌分子。该提案旨在利用 肠道病原体控制其宿主中炎症相关信号传导途径的进化能力， 使细菌效应分子适合作为治疗剂。实现治疗潜力的一个主要挑战是 这些分子能够设计一种能够将蛋白质输送到肠道内的输送系统， 上皮细胞该提案的目的是创造具有以下能力的细菌蛋白质纳米颗粒： 递送细菌效应蛋白，抑制上皮炎症，并减轻IBD的症状。关键 结果是：（1）一种新的，长期需要的IBD治疗，在源头上阻止炎症;（2）一种新的， 利用细菌免疫调节机制并设计纳米颗粒的治疗范例 临床可行性所必需的递送策略。确定了三个具体目标： 目标1。设计纳米颗粒的细胞摄取和运输特性，以最大限度地提高蛋白质 交付.细菌蛋白质纳米颗粒将被制造成具有不同的理化特征，包括大小， 交联密度、靶向配体和内体逃逸基序。暴露于纳米颗粒的上皮细胞 将在体外评估颗粒内化、内体逃逸和解离成可溶性蛋白。 目标2.评估对细菌蛋白纳米颗粒的体外和离体生物反应。各种 纳米颗粒制剂，包括来自目标1的那些以及在该目标中优化的用于运输的颗粒 通过粘液，将被施加到非极化和极化的上皮细胞，以及离体粘膜制剂 在一定的剂量范围内功能活性将通过粘膜摄取、免疫抑制和免疫抑制来测量。 信号通路，以及降低的炎性细胞因子水平。 目标3.确定优化用于体内递送的蛋白质纳米颗粒对肿瘤细胞的治疗效果。 生病的动物纳米颗粒将通过表面涂层进行改性，以增加其穿过纳米颗粒的能力。 胃肠道和目标发炎粘膜。这些颗粒将被施用至具有诱导的 结肠炎的化学、免疫学和遗传学模型以及健康对照。临床参数，粘膜 和全身炎症标志物，以及组织病理学将在相关时间点进行跟踪。

Bioengineering bacterially derived immunomodulants: a novel therapeutic approach to IBD Project Summary The long term goal is to develop novel, effective therapeutics that harness the immunomodulatory properties of bacterial molecules for the treatment of inflammatory bowel disease (IBD). The proposal aims to exploit the evolved ability of intestinal pathogens to control inflammatory related signaling pathways in their host, by adapting bacterial effector molecules as therapeutics. A major challenge in realizing the therapeutic potential of these molecules is the ability to engineer a delivery system capable of delivering protein inside intestinal epithelial cells. The objective of this proposal is to create bacterial protein nanoparticles with the ability to deliver bacterial effector proteins, suppress epithelial inflammation, and attenuate the symptoms of IBD. Key outcomes are: (1) a new, long-needed IBD therapeutic that arrests inflammation at the source; (2) a new therapeutic paradigm that utilizes bacterial immunoregulatory mechanisms and engineers a nanoparticle delivery strategy essential for clinical viability. Three specific aims have been set: Aim 1. Engineer the cellular uptake and trafficking properties of nanoparticles to maximize protein delivery. Bacterial protein nanoparticles will be fabricated with different physiochemical features including size, crosslinking density, targeting ligands, and endosomal escape motifs. Epithelial cells exposed to nanoparticles in vitro will be assessed for particle internalization, endosomal escape, and disassociation into soluble protein. Aim 2. Assess in vitro and exvivo biological response to bacterial protein nanoparticles. Various nanoparticle formulations, including those from Aim 1 as well as particles optimized in this aim for transport through mucus, will be applied to unpolarized and polarized epithelial cells, and ex vivo mucosal preparations at a range of dosages. Functional activity will be measured by mucosal uptake, suppression of immune signaling pathways, and reduced levels of inflammatory cytokines. Aim 3. Determine the therapeutic effect of protein nanoparticles optimized for in vivo delivery on diseased animals. Nanoparticles will be modified via surface coatings to increase their ability to traverse the gastrointestinal tract and target inflamed mucosa. These particles will be administered to mice with induced chemical, immunological and genetic models of colitis and to healthy controls. Clinical parameters, mucosal and systemic inflammatory markers, and histopathology will be tracked over relevant time points.