Molecular Pharmacology of NKT Cell Agonists

描述(由申请人提供)： 如果我们学会如何在体内操纵NKT细胞，它将提供巨大的治疗潜力。作为免疫的细胞佐剂，NKT细胞启动并控制感染环境中树突状细胞的成熟。因此，这些细胞的招募可能对疫苗开发、慢性传染病的免疫治疗、预防感染和癌症疫苗至关重要。然而，成功将在很大程度上取决于我们将小鼠NKT生物学知识转化为人类的能力。例如，能够激活NKT细胞的化合物的药理学几乎没有被探索，细胞的摄取和分解代谢也是如此。由于NKT细胞的激活剂是糖脂，人类脂蛋白和清道夫受体的多态性很可能会严重影响这一新的免疫调节剂家族的效力。我们建议解决NKT激动剂在小鼠和人类中的分子药理学的一些基本方面，以帮助这些分子在医学上的成功使用。这一目标将通过3个具体目标来实现：目标1：NKT细胞激动剂转运和代谢的蛋白质组学研究。基于我们以前的工作，我们假设NKT激动剂与其他脂类相比具有独特的生物学特性。需要在NKT生物学的背景下设计和使用原始工具，以推动该领域的实质性进展及其临床研究。我们会 结合新的化学和重组蛋白质工程来获取NKT激动剂的分子药理学。目的2：NKT细胞激动剂的转运。利用传统的血清生化方法，我们已经分离并鉴定了FAAH和GalCer之间的关联功能。使用蛋白质组学方法，我们现在已经对所有GalCer相关蛋白的血清进行了分析。我们假设这些蛋白质中的许多将对NKT激动剂的生物学产生积极或消极的影响。这些蛋白质的研究将使用一个强大的实验系统进行，该系统结合了重组蛋白质表达、体外生物物理和功能研究以及体内疫苗接种模型系统，然后再转移到人类细胞。目的3：NKT细胞激动剂的细胞摄取和分解代谢。NKT激动剂的摄取、加工和分解代谢仅被粗略地研究过。我们将把蛋白质组学研究与新的生物化学和细胞生物学方法结合起来，以了解NKT抗原是如何运送和处理到CD1d装载间隔的。随着NKT激动剂迅速进入临床领域，我们将通过目前的建议获得的知识将是成功的翻译研究的关键。

DESCRIPTION (provided by applicant): NKT cells offer an enormous therapeutic potential if we learn how to manipulate them in vivo. As cellular adjuvants of immunity, NKT cells prime and control the maturation of dendritic cells in al infectious contexts. As such, the recruitment of these cells could be critical to vaccine development, immunotherapy of chronic infectious diseases, prevention of infection, and cancer vaccines. However, success will rely heavily on our ability to translate the knowledge of mouse NKT biology to humans. For instance, the pharmacology of compounds capable of activating NKT cells is almost unexplored as are cellular uptake and catabolism. Since, activators of NKT cells are glycolipids, it is most likely that lipoprotein and scavenger receptor polymorphism in human will heavily influence the potency of this new family of immunomodulators. We propose to address some fundamental aspects of the molecular pharmacology of NKT agonists in mice and humans to help a successful use of those molecules in medicine. This goal will be attained through 3 specific aims: Aim 1: Proteomic studies of NKT cell agonist transport and metabolism. Based on our previous work, we hypothesize that NKT agonists have a unique biology when compared to other lipids. Original tools need to be designed and used in the context of NKT biology to drive substantial progress in the field and its translation to clinical studies. We will combine novel chemistry and recombinant protein engineering to access the molecular pharmacology of NKT agonists. Aim 2: Transport of NKT cell agonists. Using traditional serum biochemistry, we have isolated and characterized the function of the association between FAAH and ¿GalCer. Using proteomic approaches we have now profiled serum for all ¿GalCer-associated proteins. We hypothesize that many of these proteins will impact positively or negatively the biology of NKT agonists. The study of each of these proteins will be carried out using a robust experimental system combining recombinant protein expression, in vitro biophysical and functional studies, and in vivo model systems of vaccination before being translated to human cells. Aim 3: Cellular uptake and catabolism of NKT cell agonists. Uptake, processing and catabolism of NKT agonists have been only superficially examined. We will combine proteomics studies with novel biochemistry and cell biology approaches to understand the delivery and processing of NKT antigens to CD1d loading compartments. As NKT agonists are entering rapidly the clinical field, the knowledge that we will acquire through the current proposal will be critical for successful translational studies.