Enhancing immune mediated head and neck cancer anti-tumor activity using nanoparticles

摘要 对于肿瘤为 HPV 阴性的头颈癌患者，目前的治疗不会导致症状 寿命长，大多数死于原发肿瘤的局部区域复发。我们发现 HPV(-) 头颈癌患者根据其 PD-L1 和 CD8 状态分为四个不同的类别。 这些类别具有离散的结果，例如局部区域复发和疾病特异性生存率， CD8 和 PDL-1 双阳性患者的情况比任何其他患者队列更差。我们询问是否可以取消 制定可以利用这些患者类别的独特生物学特性的治疗策略。我们假设 PDL-1 的存在要么阻止 CD8 细胞毒性 T 细胞的浸润，要么导致 T 细胞耗竭，从而使 T 细胞 它到达肿瘤实质。进一步假设，我们询问表面 PDL-1 的过度表达是否可以用作 放射增敏金纳米粒子的归巢机制。我们还询问阻断 PDL-1 是否可以复制 PDL-1 阴性队列的肿瘤反应。最后，我们考虑了可以增加免疫原性的策略 缺乏 PDL-1 的肿瘤。在这个提案中，我们提出三个主要问题：1）我们可以使用 PDL-1 来存储黄金吗？ 纳米粒子并特别增强肿瘤细胞内的辐射？我们假设过度表达 PDL-1 可能作为放射增敏的理想归巢策略。金纳米粒子有很好的描述 放射增敏剂很容易通过抗体靶向肿瘤。我们将共轭金纳米粒子球体 α-PDL-1 并检查其体内肿瘤控制和总体生存的功效。 2）通过什么机制 PDL-1 阻断浸润性细胞毒性 T 细胞的活性，这种机制能否被推翻？使用 头颈癌的同基因模型，我们将研究是否通过基因操作消除 PDL-1 或 抑制性抗体可以改善基因毒性治疗后的肿瘤控制。我们将进一步分析 T 细胞 从人体组织和动物肿瘤中渗透，并询问 PDL-1 进入后轮廓是否发生变化 抑制。目的是了解肿瘤细胞中 PDL-1 的过度表达是否与以下疾病相关或只是其原因： 全身免疫抑制。 3）辐射可以增加垂死肿瘤细胞的免疫原性吗？我们呈现 数据证明具有高线性传递能量（LET）的辐射可以通过粒子实现 治疗，增加体外肿瘤细胞的免疫原性信号。因此，我们将调查是否治疗 使用具有高LET的质子束可以在体内达到类似的效果。最后，作为三击策略的测试， 我们将研究通过将 α-PDL-1 标记的金纳米粒子与 高LET质子辐射。我们预计这种治疗将阻止 T 细胞耗竭，增强放射效果 感染，并增加垂死肿瘤细胞的免疫原性。我们对肿瘤之间相互作用的理解 生物学和免疫系统表明，必须开发新的策略来整合我们的免疫系统。 将纳米技术、辐射反应和免疫学知识融入复杂和创新的治疗中 治疗策略。

ABSTRACT For patients with head and neck cancer whose tumors are HPV negative, current therapy does not lead to sig- nificant longevity and most succumb to loco-regional recurrence of the primary tumor. We discovered that HPV(-) head and neck cancer patients fit into four distinct categories relative to their PD-L1 and CD8 status. These categories had discrete outcomes such as loco-regional recurrence and disease-specific survival, with CD8 and PDL-1 double positive patients faring worse than any other patient cohort. We asked if we can de- velop treatment strategies that can leverage the unique biology of these patient categories. We posit that pres- ence of PDL-1 either blocks infiltration of CD8+ cytotoxic T cells or leads to exhaustion of T cells that do make it to the tumor parenchyma. Postulating further, we asked if overexpression of surface PDL-1 can be used as a homing mechanism for radiosensitizing gold nanoparticles. We also asked if blocking PDL-1 could replicate the tumor response of PDL-1 negative cohorts. Finally, we considered strategies that can increase immunogenicity of tumors that lack PDL-1. In this proposal, we ask three main questions: 1) Can we use PDL-1 to home gold nanoparticles and enhance radiation specifically within tumor cells? We hypothesized that overexpression of PDL-1 may serve as the ideal homing strategy for radiosensitization. Gold nanoparticles are well described radiosensitizers that are easily targeted to tumors via antibodies. We will conjugate spheroid gold nanoparticles to α-PDL-1 and examine their efficacy in vivo for tumor control and overall survival. 2) By what mechanism is PDL-1 blocking the activity of infiltrating cytotoxic T cells and can this mechanism be overturned? Using a syngeneic model of head and neck cancer, we will investigate if eliminating PDL-1 by genetic manipulation or an inhibitory antibody can improve tumor control following genotoxic therapy. We will further profile the T cell infiltrate from both the human tissues and the animal tumors and ask if the profile changes following PDL-1 in- hibition. The goal is to understand if PDL-1 overexpression in tumor cells is simply correlative to or causative of the systemic immune inhibition. 3) Can radiation increase immunogenicity of dying tumor cells? We present data to demonstrate that radiation with high linear transfer energy (LET), which can be achieved by particle therapy, increased immunogenic signals from tumor cells in vitro. Therefore, we will investigate if treatment with proton beams with high-LET can achieve a similar effect in vivo. Finally, as a test of a three-hit strategy, we will examine the tumor control that can be achieved by combining α-PDL-1 tagged gold nanoparticles with high-LET proton radiation. We anticipate that this treatment will block T-cell exhaustion, enhance radiation ef- fects, and increase the immunogenicity of dying tumor cells. Our understanding of the interplay between tumor biology and the immune system suggests that new strategies will have to be developed that can merge our knowledge of nanotechnology, radiation response, and immunology into sophisticated and innovative treat- ment strategies.