Development of a lipid-based platform for mucosal delivery and mucosal vaccination

项目摘要 该提案的目标是测试鞘糖脂运输的生物学是否可以用于 治疗性肽和抗原在体内穿过紧密的粘膜屏障的递送。 目前，还没有有效的方法用于口服药物递送穿过内衬于胃粘膜的紧密粘膜上皮屏障。 肠子在这里，我们提出了一种基于脂质的粘膜递送平台的研究， 利用内源性基于糖脂的跨膜转运途径的治疗肽和抗原 极化上皮细胞这种途径将短的或不饱和的GM1-神经酰胺在细胞内分类， 基底外侧膜-从而通过胞吞作用突破上皮屏障。我们的目标是设计GM1载体 使治疗物质内化并有效地穿过肠上皮屏障。我们最近 鉴定的转胞吞作用后从细胞膜有效释放的GM1种类和另一组 GM1物种不能有效释放，而是保留在膜上更长的时间。我们将 通过粘膜施用融合至细胞的非天然GM1种类来扩展我们的原理研究证据。 肠促胰岛素激素GLP-1作为治疗II型糖尿病模型。进一步的结构-功能研究 GM1的细胞外寡糖结构域和使该技术成为可能的机制研究也是一个挑战。 当前提案的重点。GM1平台将适用于多种药物的粘膜递送。 大分子和小分子药物以及治疗性蛋白质和肽-包括用于免疫的粘膜抗原 系统调制然后，我将测试GM1物种是否可以允许抗原的递送（以卵清蛋白为模型） 穿过粘膜屏障，例如能够诱导免疫耐受（或粘膜接种）。 我将利用我们独特的试剂和研究方法，以及整个Lencer实验室的可用资源， 和朗伍德医学校园，以及我们已经召集的专家科学家小组， 我提出了实验，发展我的职业生涯作为一个独立的研究人员，并实现我的目标，使 对脂质生物学和疫苗开发领域的新贡献。

PROJECT SUMMARY The goal of this proposal is to test whether the biology of glycosphingolipid trafficking can be harnessed for delivery of therapeutic peptides and antigens across tight mucosal barriers in vivo. Currently, there are no effective methods for oral drug delivery across tight mucosal epithelial barriers lining the intestine. Here, we propose studies towards the development of a lipid-based platform for mucosal delivery of therapeutic peptides and antigens exploiting an endogenous glycoshingolipid-based trafficking pathway across polarized epithelial cells. This pathway sorts short- or unsaturated GM1-ceramides across the cell to the basolateral membrane - thus breeching the epithelial barrier by transcytosis. Our aim is to design GM1 carriers that internalize therapeutic cargoes and efficiently cross the intestinal epithelial barrier. We have recently identified GM1 species that are efficiently released from cell membranes after transcytosis and another set of GM1 species that are not efficiently released, but are rather retained on membranes for longer periods. We will expand our proof of principle studies by mucosal administration of the non-native GM1 species fused to the incretin hormone GLP-1, as model for treatment of Type II Diabetes. Further structure-function studies of the extracellular oligosaccharide domain of GM1 and mechanistic studies to enable the technology are also a focus of the current proposal. The GM1 platform would be applicable to the mucosal delivery of a wide range of large and small molecule drugs and therapeutic proteins and peptides- including mucosal antigens for immune system modulation. I will then test if GM1 species can allow for delivery of antigens (modeled by ovalbumin) across mucosal barriers such as enabling induction of immunological tolerance (or mucosal vaccination). I will leverage our unique reagents and research approach, the available resources throughout the Lencer Lab and Longwood Medical Campus, and group of expert scientists we have already assembled to complete the proposed experiments and develop my career as an independent researcher and achieve my goal to make novel contributions to the field of lipid biology and vaccine development.