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信号传导， 寿命很长，大多数死于原发肿瘤的局部复发。我们发现 HPV（-）头颈癌患者根据其PD-L1和CD 8状态分为四个不同的类别。 这些类别有离散的结果，如局部区域复发和疾病特异性生存， CD 8和PDL-1双阳性患者的情况比任何其他患者队列更差。我们问能不能- velop治疗策略可以利用这些患者类别的独特生物学。我们把它- PDL-1的存在或者阻断CD 8+细胞毒性T细胞的浸润，或者导致T细胞的耗竭， 转移到肿瘤实质。进一步假设，我们询问表面PDL-1的过表达是否可以用作 放射增敏金纳米粒子的归巢机制。我们还询问了阻断PDL-1是否可以复制 PDL-1阴性组群的肿瘤反应。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。在这份提案中，我们提出三个主要问题：1）我们能用PDL-1来安家黄金吗 纳米粒子和增强辐射专门在肿瘤细胞内？我们假设， PDL-1可作为理想的放射增敏归巢策略。金纳米颗粒被很好地描述 容易通过抗体靶向肿瘤的放射增敏剂。我们将球状金纳米粒子 与α-PDL-1结合，并检查它们在体内肿瘤控制和总生存期的功效。2)通过什么机制 PDL-1阻断浸润性细胞毒性T细胞的活性，这种机制可以被推翻吗？使用 头颈癌的同基因模型，我们将研究是否通过基因操作或 抑制性抗体可以改善遗传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浸润，并询问PDL-1后的轮廓是否发生变化- - 是的目的是了解肿瘤细胞中PDL-1的过度表达是否仅仅与肿瘤相关或导致肿瘤的发生。 全身免疫抑制。3)放射能增加垂死肿瘤细胞的免疫原性吗？我们提出 数据表明，具有高线性转移能量（LET）的辐射，可以通过粒子实现 治疗，增加来自体外肿瘤细胞的免疫原性信号。因此，我们将研究是否治疗 用高LET的质子束可以在体内达到类似的效果。最后，作为一个三击战略的测试， 我们将研究通过结合α-PDL-1标记的金纳米颗粒与 高LET质子辐射。我们预计，这种治疗将阻止T细胞耗竭，增强辐射效应， 并增加垂死肿瘤细胞的免疫原性。我们对肿瘤和癌症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， 将纳米技术、辐射反应和免疫学知识转化为复杂和创新的治疗方法， 战略。

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.