Designing a targeted 'super adjuvant' nanoparticle platform for vaccination

项目摘要 这项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 预防针疫苗由2种成分组成：由CD 8+细胞毒性T细胞识别的肿瘤特异性抗原 细胞（CTL）和佐剂，所述佐剂为抗原呈递细胞提供必要的共刺激细胞因子信号。 细胞（APC）来引发和激活CTL应答。然而，在像癌症这样的复杂疾病中， 佐剂疫苗可能不能完全有效地减轻免疫抑制剂的无数免疫抑制作用， 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”制剂中共同递送多种佐剂可以利用 多种模式识别受体途径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，其中预先选择的 将抗原和佐剂递送至淋巴结，原位接种，其中仅递送佐剂 直接作用于肿瘤，有明显的优势。在原位方法中，肿瘤本身在肿瘤细胞中提供抗原。 从垂死的肿瘤细胞脱落的新抗原的形式。这种方法在一个 在个体化的基础上，并且不需要肿瘤抗原的先验知识。目前的现场方法， 包括游离激动剂、游离细胞因子和免疫原性病毒，严重依赖于直接瘤内注射， 使脱靶毒性最小化，并且是癌症类型特异性的。瘤内递送本身也有严重的局限性 在传递和功效方面。在我们最近的出版物和额外的初步数据的支持下，这一高- 一项风险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， 纳米颗粒（免疫NP）平台，共封装3个协同I型干扰素驱动的先天免疫 使用基于脂质的材料和微流体技术在相同的NP上使用激动剂。具体而言，60-nm PEG化 免疫-NP将被设计为在全身血液循环中安全地递送，以共同递送以下激动剂： STING、TLR 4和TLR 9通路。它们还将靶向多种特定类型的细胞， TME，包括APC，活化的内皮细胞和肿瘤细胞，以协调一致的多细胞 这可能是消除异质性侵袭性肿瘤所必需的反应。我们假设目标 三激动剂“超佐剂”免疫-NP将驱动协同的I型干扰素介导的APC应答， 在多种TNBC小鼠模型中激活CTL用于肿瘤清除。特异性目标1将识别免疫NP 在促进的细胞因子宽度和深度方面的最佳功能所需的设计参数。具体目标 2将建立有效的TME归巢和治疗效果的免疫NP靶向方案。这些 研究将有效地解决严重限制当前的药物输送、疗效和安全性挑战。 接近。作为一种平台技术，这种“超级佐剂”免疫NP的成功开发也具有 广泛应用于癌症和传染病的标准淋巴结定向疫苗接种。

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.