A high-throughput nanoparticle assay to characterize cancer neoepitope-specific T cells

摘要 免疫疗法在癌症中的应用取得了令人印象深刻和鼓舞人心的结果。然而，这些 结果似乎只适用于一小部分患者。许多从业者和研究人员的目标是发现 为什么患者会有不同的反应。然而，现有的表征癌症的技术并没有集中在 免疫反应本身。因此，我们的目标是用一种直接表征 癌症在其免疫反应方面。为此，我们将使用我们实验室开发的一项核心技术， 利用磁性纳米颗粒来丰富和激活针对CD8 T细胞的癌症，以检测和， 有可能达到相同的治疗水平。具体地说，我们将寻找新的表位-由 可以产生免疫反应的癌症本身的突变。我们已经证明了这一点的可行性 在小鼠和人类的情况下都有这种方法；然而，我们还没有将这种分析发展成一种高度的- 涵盖广泛的异质人口的吞吐量方法。为此，我们将首先设计 磁性纳米颗粒被称为人工抗原提呈细胞(AAPC)，因为它们既有一种抗原- 在其表面负载人类白细胞抗原(HLA)和共刺激分子。工程参数 将包括大小、配基密度和配基选择。此外，我们还将通过开发96口井来扩大产量 平板高通量版的化验。我们还将通过开发技术来扩大这项技术的覆盖范围 AAPC用于额外的I类HLA等位基因，也用于AAPC用于CD4T细胞刺激。最后，通过我们的 与纽约大学的Jeff Weber博士合作，我们将通过测量和检测来验证这项技术 来自IV期黑色素瘤患者的新表位。我们将评估治疗如何影响免疫反应 在检查点阻断治疗前后对肿瘤进行探查。这项技术将填补提供 癌症的免疫学特征在两种小鼠模型和癌症患者的危急情况下。这 代表性将塑造治疗的提供方式和下一代治疗的方式 发展起来的。

ABSTRACT The application of immunotherapy to cancer has yielded impressive and inspiring results. However, these results seem to only apply to a subset of patients. Many practitioners and researchers are aiming to discover why there are differential responses from patients. Yet existing techniques to characterize cancer do not focus on the immune response itself. Therefore, we aim to fill this gap with a technology that directly characterizes the cancer in terms of its immune response. To do this, we will use a core technology, developed in our labs, that utilizes magnetic nanoparticles to enrich and activate cancer targeting CD8+ T cells to detectable and, potentially even, therapeutic levels. Specifically, we will look for neoepitopes—epitopes generated by the mutations of the cancer itself that can generate an immune response. We have demonstrated feasibility of this approach within both murine and human contexts; however, we have yet to develop the assay into a high- throughput approach that covers a broad heterogenous human population. To do so, we will first engineer the magnetic nanoparticles, called artificial antigen-presenting cells (aAPCs) because they have both an antigen- loaded human leukocyte antigen (HLA) and co-stimulatory molecules on their surface. Engineering parameters will include size, ligand density, and ligand choice. Furthermore, we will extend output by developing a 96-well plate high-throughput version of the assay. We will also broaden the reach of this technology by developing aAPCs for additional class I HLA alleles and also for an aAPC for CD4+ T cell stimulation. Finally, through our collaborative efforts with Dr. Jeff Weber at NYU, we will validate the technology by measuring and detecting neoepitopes from stage IV melanoma patients. We will assess how treatment affects the immune response to the tumor by probing before and after checkpoint blockade therapy. This technology will fill the gap of providing an immunological characterization of cancer in both murine models and critically in patients with cancer. This representation will shape both how therapy is delivered and how the next generation of therapies are developed.