Unsaturated glycoceramide as novel platform for mucosal vaccine and drug delivery

不饱和甘油酰胺作为粘膜疫苗和药物输送的新平台

基本信息

批准号：
8145606
负责人：
WAYNE I LENCER
金额：
$ 21.51万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-30 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8145606
关键词：
Address Adjuvant Agonist Antigens Apical Area Asthma Biological Assay Biology Blood Glucose Cell surface Cells Ceramides Cholera Toxin Dendritic Cells Destinations Development Drug Delivery Systems Endocytosis Endosomes Environment Epithelial Epithelial Cells Fatty Acids Flagellin Ganglioside GM1 Gene Expression Goals Hormones Immune In Vitro Inflammatory Bowel Diseases Injection of therapeutic agent Intestines Invaded Lamina Propria Leptin Life Ligands Link Lipids Lysosomes Measurable Measures Membrane Protein Traffic Memory B-Lymphocyte Methods Modeling Mucosal Immune Responses NF-kappa B Nose Oral Ovalbumin Pathway interactions Peptides Play Proteins Recycling Research Role Salmonella Saturated Fatty Acids Sorting - Cell Movement Structure Surface T-Lymphocyte TLR5 gene Technology Testing Therapeutic Tight Junctions Tissues Toll-like receptors Toxic effect Translating Vaccine Adjuvant Vaccine Antigen Variant Work absorption apical membrane base clinical application diabetic glucagon-like peptide glucose tolerance glycolipid receptor improved in vitro testing in vivo innovation late endosome lipid structure lipid transport molecular carrier monolayer mucosal vaccination mucosal vaccine novel pathogen peptide hormone prevent programs public health relevance response therapeutic protein therapeutic vaccine trafficking vaccine delivery

项目摘要

DESCRIPTION (provided by applicant): The goal of this proposal is to test if our recent discovery on lipid trafficking in epithelial cells can be translated to clinical applications. Specifically, we will test the utility of using "short or unsaturated" ceramide-based lipids as molecular carriers to deliver therapeutic peptides or vaccine adjuvants across mucosal epithelial barriers. Mucosal surfaces represent vast areas where host tissues are separated from the environment only by a delicate but highly effective single layer of columnar epithelial cells, joined by tight junctions that are impermeable to proteins and even small peptides. So far, the lack of rational and efficient methods to circumvent this barrier has prevented the application of most therapeutic proteins for oral or nasal drug delivery and for mucosal vaccines. In the course of our studies on the biology of the glycolipid receptor for cholera toxin, ganglioside GM1, we recently discovered that the structure of the ceramide (lipid) domain dictates GM1 trafficking in epithelial cells. When applied apically, GM1-ceramides containing "short" C12:0 or "kinked chain" unsaturated C16:1 fatty acids (GM1short/unsat) enter the common/recycling endosome. Here, they are sorted for transport to various intracellular destinations and into the "transcytotic" pathway to the basolateral cell surface. In contrast, the GM1-ceramides with fully saturated fatty acid chains (C16:0 or longer) (GM1 long/sat) are instead transported to the late endosome and lysosome for degradation. In this exploratory project, we will test whether this basic discovery can be harnessed for transepithelial delivery of a bioactive peptide or protein adjuvant, both of which have clinical applications. We will link GM1 molecules containing unsaturated "kinked" or saturated fatty acids to the therapeutic peptide hormone glucagon-like peptide-1 (GLP1), which acts to regulate blood sugar (Aim 1); and to the TLR5-agonist FliC (Salmonella flagellin), which we use here to model a mucosal vaccine adjuvant (Aim 2). These studies will test a novel platform for transport of therapeutic/vaccine molecules across mucosal surfaces. There is great need for non-parental delivery of therapeutic peptides and proteins. Improved mucosal vaccine strategies are greatly needed for protection against pathogens, the vast majority of which invade via mucosal surfaces. PUBLIC HEALTH RELEVANCE: The goal of this application is to develop a way to allow for oral or nasal administration of therapeutic proteins and vaccines. Normally such proteins are not absorbed and must be delivered by injection; severely limiting therapeutic applications.

描述（由申请人提供）：该提案的目的是测试我们最近关于上皮细胞脂质运输的发现是否可以转化为临床应用。具体来说，我们将测试使用“短或不饱和”神经酰胺基脂质作为分子载体来跨粘膜上皮屏障递送治疗肽或疫苗佐剂的效用。粘膜表面代表了广阔的区域，宿主组织仅通过精致但高效的单层柱状上皮细胞与环境分开，并通过蛋白质甚至小肽不可渗透的紧密连接连接。迄今为止，由于缺乏合理有效的方法来规避这一障碍，阻碍了大多数治疗性蛋白在口服或鼻腔药物递送以及粘膜疫苗中的应用。在我们对霍乱毒素糖脂受体神经节苷脂 GM1 生物学的研究过程中，我们最近发现神经酰胺（脂质）结构域的结构决定了 GM1 在上皮细胞中的运输。当顶部应用时，含有“短”C12:0 或“扭结链”不饱和 C16:1 脂肪酸（GM1short/unsat）的 GM1-神经酰胺进入共同/回收内体。在这里，它们被分类运输到各个细胞内目的地并进入到基底外侧细胞表面的“转胞吞”途径。相比之下，具有完全饱和脂肪酸链（C16:0 或更长）（GM1 long/sat）的 GM1-神经酰胺则被转运至晚期内体和溶酶体进行降解。在这个探索性项目中，我们将测试这一基本发现是否可以用于生物活性肽或蛋白质佐剂的跨上皮递送，这两者都具有临床应用。我们将含有不饱和“扭结”或饱和脂肪酸的 GM1 分子与治疗性肽激素胰高血糖素样肽-1 (GLP1) 连接起来，其作用是调节血糖（目标 1）；以及 TLR5 激动剂 FliC（沙门氏菌鞭毛蛋白），我们在这里使用它来模拟粘膜疫苗佐剂（目标 2）。这些研究将测试一种用于跨粘膜表面运输治疗/疫苗分子的新平台。非常需要治疗性肽和蛋白质的非亲本递送。非常需要改进的粘膜疫苗策略来预防病原体，其中绝大多数病原体通过粘膜表面侵入。公共健康相关性：本申请的目标是开发一种允许口服或鼻腔施用治疗性蛋白质和疫苗的方法。通常这些蛋白质不会被吸收，必须通过注射来输送；严重限制了治疗应用。