Engineered Optimal Adjuvant and Antigen Release from Biodegradable Nanoparticles

描述（由申请人提供）：我们的目标是设计释放与肿瘤特异性抗原共同包裹的佐剂，用于黑色素瘤的免疫治疗。具体目标是：(1)通过制备胞嘧啶-磷硫酸-鸟嘌呤寡脱氧核苷酸(CpG)-抗原融合偶联物和双层颗粒，使抗原共价附着在颗粒表面，CpG包裹在颗粒基质中，从而设计出增强免疫治疗反应的可生物降解纳米颗粒。在优化颗粒配方方法后，颗粒疫苗将通过定量细胞因子/趋化因子的产生以及使用RT-PCR和ELISA检测toll样受体9 （TLR9）、IFN-3、IL- 6和IL-12的上调和下调来表征免疫应答。(2)通过确定产生最大免疫反应的给药方案（给药途径和剂量），评估小鼠黑色素瘤模型免疫治疗的最佳临床前疫苗策略。然后，优化的纳米颗粒疫苗将与其他免疫治疗方法进行测试。提出的研究代表了纳米颗粒工程的一种新方法，提供佐剂和抗原的最佳释放动力学，以刺激有效的抗癌免疫治疗反应。针对黑色素瘤的强效免疫治疗疫苗的开发将建立一个框架，可应用于一系列其他癌症模型，包括淋巴瘤、肾癌、结肠癌和胰腺癌。

DESCRIPTION (provided by applicant): Our objective is to engineer the release of adjuvant co-entrapped with tumor specific antigens for immunotherapeutic treatment of melanoma. The specific aims are: (1) Engineer biodegradable nanoparticles for enhanced immunotherapeutic responses by preparing cytosine-phosphorothiolate-guanine oligodeoxynucleotides (CpG)-antigen fusion conjugates and double layered particles with antigens covalently attached on the surface of the particles and CpG entrapped within the particle matrix. Following optimization of the particle formulation methodology, the immune response initiated by the particle vaccines will be characterized by quantifying the cytokine/chemokine production and upregulation and downregulation of Toll-like receptor 9 (TLR9), IFN-3, IL- 6 and IL-12 using RT-PCR and ELISA assays. (2) Evaluate the optimal pre-clinical vaccine strategy for immunotherapeutic treatment in a murine melanoma model by determining the dosing regimen (route of administration and number of doses) that produces maximal immune response. The optimized nanoparticle vaccine will then be tested against alternative immunotherapeutic treatments. The proposed research represents a novel approach of nanoparticle engineering to provide the optimal release kinetics of adjuvant and antigens in stimulating potent immunotherapeutic responses against carcinomas. The development of a potent immunotherapeutic vaccine against melanoma will establish a framework which can be applied to a range of other cancer models including lymphoma, renal carcinoma, colon cancer and pancreatic cancer. PUBLIC HEALTH RELEVANCE: Our objective is to engineer the release of adjuvant co-entrapped with tumor specific antigens for immunotherapeutic treatment of melanoma. The specific aims are: (1) Engineer biodegradable nanoparticles for enhanced immunotherapeutic responses by preparing cytosine-phosphorothiolate-guanine oligodeoxynucleotides (CpG)-antigen fusion conjugates and double layered particles with antigens covalently attached on the surface of the particles and CpG entrapped within the particle matrix. (2) Evaluate the optimal pre-clinical vaccine strategy for immunotherapeutic treatment in a murine melanoma model by determining the dosing regimen (route of administration and number of doses) that produces maximal immune response.