Novel Biodegradable Nanoadjuvants for Immunotherapy of Pancreatic Cancer

描述(申请人提供)：目前的治疗方法未能有效地治疗胰腺癌。由于平均存活率仍不到4%，迫切需要研究替代治疗方法。该项目的总体目标是开发一种新型的纳米级疫苗递送系统，该系统将导致对胰腺癌的强大抗肿瘤反应。肿瘤相关抗原粘蛋白-1在胰腺癌中过度表达并分化为糖基化。我们将设计疫苗配方，将粘蛋白-1包裹到具有免疫调节能力的新型两亲性聚酸酐共聚体系的纳米球中。其具体目的是：(SA1)体外研究由1，6-双(对羧基苯氧基)己烷(CPH)和1，8-双(对羧基苯氧基)-3，6-二氧辛烷(CPTEG)组成的可生物降解的两亲性聚酸酐纳米球对树突状细胞(DC)的细胞毒性和激活作用；(SA2)用小鼠模型进行体内Mucin-1纳米球的研究，以确定其免疫原性和免疫调节能力。我们的方法结合了对抗原提呈细胞(APC)摄取效率的阐明，这些细胞的激活和迁移，以及适当的细胞免疫(即抗原特异性T细胞反应)的诱导。由于颗粒大小对有效的吞噬作用起着重要作用，SA1将涉及通过不同的技术制备微/纳米球：固体/油/油复乳、低温芳构化和纳米沉淀。将微/纳米球与小鼠骨髓来源的树突状细胞培养，将在MHC I和MHC II分子的背景下研究DC的细胞毒性和高效激活。在确定哪种CPTEG：CPH制剂可诱导Th1细胞因子后，将在小鼠模型上对这些制剂进行评估，以确定有助于诱导抗肿瘤免疫的配方。纳米球佐剂的免疫反应途径和免疫调节能力(即Th1和Th2)将通过产生的细胞因子谱和抗体进行评估。基于所获得的结果，将评估野生型(C57BL/6)小鼠的抗肿瘤反应。该项目的预期结果是确定CPTEG：CPH微球/纳米球配方，用于在转基因小鼠模型中进一步测试，以评估耐受和自身免疫背景下的抗肿瘤免疫，这将导致基于疫苗的免疫疗法。关联性。这项技术的开发和应用将促进抗肿瘤疫苗的合理设计，使其能够适当调节免疫反应，发展保护性免疫，并导致抗肿瘤免疫疗法的技术革命。

DESCRIPTION (provided by applicant): Current medical therapies have failed to treat pancreatic cancer effectively. There is an urgent need to investigate alternative treatments as the average survival rate remains less than 4%. The overall objective of this project is to develop a novel nanoscale vaccine delivery system that will lead to strong anti-tumor responses against pancreatic cancer. The tumor-associated antigen Mucin-1 is over expressed and differentially glycosylated by pancreatic adenocarcinomas. We will design vaccine formulations that encapsulate Mucin-1 into nanospheres of a novel amphiphilic polyanhydride copolymer system with immunomodulatory capabilities. The specific aims are: (SA1) In vitro studies to determine cytotoxicity and activation of dendritic cells (DCs) by biodegradable amphiphilic polyanhydride nanospheres composed of 1,6-bis-(p-carboxyphenoxy)hexane (CPH) and 1,8-bis-(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG); and (SA2) In vivo studies of Mucin-1 loaded nanospheres using murine models to determine their immunogenicity and immunomodulatory capabilities. Our approach combines the elucidation of the uptake efficiency of antigen presenting cells (APCs), the activation and migration of these cells, and the induction of appropriate cell-mediated immunity (i.e. antigen-specific T cell responses). Since particle size plays a major role for efficient phagocytosis, SA1 will involve the fabrication of micro/nanospheres by different techniques: solid/oil/oil double emulsion, cryogenic aromization, and nanoprecipitation. Culture of the micro/nanospheres with murine bone marrow derived dendritic cells will investigate the cytotoxicity and efficient activation of DCs in the context of MHC I and MHC II molecules. After determination of which CPTEG:CPH formulation induce Th1 cytokines, these will be evaluated on murine models to identify formulations that will facilitate the induction of anti-tumor immunity. The immune response pathway and the immunomodulatory capabilities of the nanosphere adjuvants (i.e., Th1 vs. Th2) will be evaluated by means of the cytokine profiles and antibodies produced. Based on the results obtained, an anti-tumor response will be evaluated in wild type (C57BL/6) mice. The expected outcome of this project is the determination of CPTEG:CPH micro/nanosphere formulations for further testing in transgenic murine models to evaluate anti-tumor immunity in the context of tolerance and auto-immunity, which will lead to vaccine-based immunotherapies. Relevance. Development and application of this technology will facilitate the rational design of anti-tumor vaccines with the ability to appropriately modulate the immune response to develop protective immunity and lead to technologies that will revolutionize anti-tumor immunotherapies.