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

项目总结 这项提案的目标是测试糖鞘磷脂贩运的生物学是否可以被利用来 体内跨越紧密粘膜屏障的治疗性多肽和抗原的输送。 目前，还没有有效的方法跨越紧密的粘膜上皮屏障来给药。 肠子。在这里，我们建议研究开发基于脂质的粘膜给药平台。 利用内源性糖脂转运途径的治疗性多肽和抗原 极化的上皮细胞。这一途径通过细胞将短的或不饱和的GM1神经酰胺分选到 基侧膜--通过细胞穿透形成上皮屏障。我们的目标是设计GM1运输船 将治疗性货物内化，并有效地穿过肠道上皮屏障。我们最近做了 已鉴定的GM1物种在细胞转运后有效地从细胞膜中释放出来，以及另一组 GM1物种不能有效地释放，但可以在膜上保留更长的时间。我们会 扩展我们的原理证明研究，通过粘膜给药非本地GM1物种融合到 胰岛素激素GLP-1，作为治疗II型糖尿病的模型。血管紧张素转换酶的进一步结构功能研究 GM1的胞外低聚糖结构域和实现该技术的机制研究也是一项 当前提案的重点。GM1平台将适用于广泛的粘膜给药 大小分子药物和治疗性蛋白质和多肽--包括用于免疫的粘膜抗原 系统调制。然后，我将测试GM1物种是否可以传递抗原(以卵清蛋白为模型) 跨越粘膜障碍，例如能够诱导免疫耐受(或粘膜疫苗接种)。 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 和朗伍德医学院，以及我们已经召集的专家科学家小组，完成了 提出实验，发展我的独立研究事业，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.