Designing a targeted 'super adjuvant' nanoparticle platform for vaccination

项目总结 该提案旨在开发一种针对原位癌的靶向三激动剂“超级佐剂”纳米颗粒平台 接种疫苗。疫苗由两部分组成：一种被CD8+细胞毒性T细胞识别的肿瘤特异性抗原 细胞(CTL)和为抗原呈递提供必要的共刺激细胞因子信号的佐剂 细胞(APC)启动并激活CTL反应。然而，在癌症等复杂疾病中，单一的- 佐剂疫苗可能不能完全有效地减轻A型流感病毒的无数免疫抑制效应 异质侵袭性肿瘤微环境(TME)，如三阴性乳腺癌 (TNBC)。在合理设计的“超级佐剂”配方中联合使用多种佐剂可以利用 多个模式识别受体通路同时驱动促炎协同细胞因子 既有广度又有深度的回应。此外，与标准疫苗接种相比，在标准疫苗接种中， 抗原和佐剂被输送到淋巴结，即原位疫苗接种，在那里只输送佐剂 直接对肿瘤，有明显的优势。在一种原位方法中，肿瘤本身提供了 新抗原的形式从死亡的肿瘤细胞中脱落。这种方法捕获患者在 个性化的基础上，不需要肿瘤抗原的先验知识。目前的就地方法， 包括游离激动剂、游离细胞因子和免疫原性病毒在内，严重依赖肿瘤内直接注射 将偏离目标的毒性降至最低，并针对癌症类型。瘤内分娩本身也有严重的局限性。 在交付和疗效上都是如此。在我们最近的出版物和额外的初步数据的支持下，这一高位- Risk提案试图通过设计一种“超级佐剂”免疫调节剂来解决这些缺点 共包裹3种协同I型干扰素驱动天然免疫的纳米颗粒(免疫-NP)平台 使用脂基材料和微流体在同一NP上使用激动剂。具体来说，60 nm聚乙二醇化 免疫纳米粒将被设计成在全身血液循环中安全地递送，以共同递送激动剂 肿瘤的刺痛、TLR4和TLR9通路。它们还将针对多种特定类型的细胞 TME，包括APC、激活的内皮细胞和肿瘤细胞，协调协调多细胞 可能是消除异质侵袭性肿瘤所必需的反应。我们假设目标是 三激动剂“超级佐剂”免疫纳米粒将驱动I型干扰素介导的协同APC反应， 在多种TNBC小鼠模型中激活CTL以清除肿瘤。特定目标1将识别免疫NP 在促进细胞因子的广度和深度方面，最佳功能所需的设计参数。特定目标 2将建立免疫-NP靶向计划，以有效地进行TME归巢和治疗效果。这些 研究将有效地解决严重限制电流的交付、有效性和安全性挑战 接近了。作为一种平台型技术，这种免疫核糖核酸的成功研发也具有 广泛应用于癌症和传染病的标准淋巴导向疫苗接种。

PROJECT SUMMARY This proposal seeks to develop a targeted tri-agonist “super adjuvant” nanoparticle platform for in situ cancer vaccination. Vaccines consist of 2 components: a tumor-specific antigen that is recognized by CD8+ cytotoxic T cells (CTLs) and an adjuvant that provides the necessary costimulatory cytokine signals to antigen-presenting cells (APCs) to prime and activate a CTL response. In complex diseases like cancer, however, a single- adjuvant vaccine may not be fully effective to mitigate the myriad immunosuppressive effects of a heterogeneous aggressive tumor microenvironment (TME) such as that of triple-negative breast cancer (TNBC). Co-delivery of multiple adjuvants in a rationally designed “super adjuvant” formulation can harness multiple pattern recognition receptor pathways simultaneously to drive a proinflammatory synergistic cytokine response that has both breadth and depth. Further, compared to standard vaccination, where a preselected antigen and adjuvant are delivered to lymph nodes, in situ vaccination, where only the adjuvant is delivered directly to the tumor, has clear advantages. In an in situ approach, the tumor itself provides the antigen in the form of neoantigens shed from dying tumor cells. This approach captures patient heterogeneity on a personalized basis and does not require prior knowledge of tumor antigens. Current in situ approaches, including free agonists, free cytokines, and immunogenic viruses, rely heavily on direct intratumoral injection to minimize off-target toxicity and are cancer type-specific. Intratumoral delivery itself also has serious limitations in both delivery and efficacy. Supported by our recent publications and additional preliminary data, this high- risk proposal seeks to address these shortcomings by designing a “super adjuvant” immunomodulatory nanoparticle (immuno-NP) platform that coencapsulates 3 synergistic Type I interferon-driving innate immune agonists on the same NP using lipid-based materials and microfluidics. Specifically, 60-nm PEGylated immuno-NPs will be designed to be delivered safely in the systemic blood circulation to co-deliver agonists of the STING, TLR4, and TLR9 pathways to tumors. They will also be targeted to multiple specific types of cells in the TME, including APCs, activated endothelial cells, and tumor cells, to orchestrate a concerted multi-cellular response that may be necessary to eliminate heterogeneous aggressive tumors. We hypothesize that targeted tri-agonist “super adjuvant” immuno-NPs will drive a synergistic Type I interferon-mediated APC response that activates CTLs for tumor clearance in multiple mouse models of TNBC. Specific Aim 1 will identify immuno-NP design parameters required for optimal function in terms of promoted cytokine breadth and depth. Specific Aim 2 will establish immuno-NP targeting schemes for effective TME homing and therapeutic efficacy. These studies will effectively address the delivery, efficacy, and safety challenges that severely limit current approaches. As a platform technology, successful development of this “super adjuvant” immuno-NP also has wide-ranging applications in standard lymph node-directed vaccination for both cancer and infectious disease.