Probing cellular, molecular and biomechanical barriers to immunotherapy in the tumor microenvironment with organotypic in vitro models of the tumor-lympho-immune interface

项目摘要/摘要 免疫疗法有望治愈许多癌症和转移性疾病，但仅限于 使一小部分患者受益。肿瘤仍然可以参与多种机制来避免和逃避抗肿瘤 免疫反应，包括抑制、灭活和排除潜在的细胞毒性T细胞的过程 它们与肿瘤微环境(TME)中的细胞合作。对这些障碍有更好的理解 导致了大量新的免疫调节靶点的开发，其中一些将与 例如，检查站封锁或CAR T细胞。另一方面，免疫治疗的主要障碍可能是 在同一癌症类型的患者中是不同的，因此需要个性化的方法来 免疫疗法，以便使用适当的靶点。在这里，我们开发了一种新型的器官型培养装置 维持原发肿瘤的体外培养和免疫成分(肿瘤引流淋巴结或 循环白细胞)，在一个平台上，能够精确地控制空间、分子、细胞和 机械特性，即相对较高的处理量，以允许筛选或大量 实验变量。在初步数据中，我们表明这些设备反映了体内的关键特征 对免疫治疗的反应，如提高肿瘤细胞杀伤率和增加免疫毒性标记物 (可能的不良事件)对细胞因子免疫治疗的反应。我们建议这些设备可以用来 既是为了筛选理想的免疫治疗组合，也是为了探索 对表现出不同程度免疫渗透的肿瘤的抗肿瘤免疫反应的缺陷， 新抗原载量和基线淋巴密度。通过这种方式，我们可以开始建立一个分层图， 将个别肿瘤的关键形态特征与最有可能导致 疗效和肿瘤消退。使用小鼠模型和原始患者来源的组合 ，我们将利用我们的新型有机型设备提供的控制水平来 机械地询问单个免疫细胞亚群和信号轴，以了解它们的 在影响抗肿瘤免疫反应的过程和结果中的作用。

PROJECT SUMMARY/ABSTRACT Immunotherapies hold immense promise to provide cures for many cancers and metastatic disease, but only benefit a fraction of patients. Tumors can still engage multiple mechanisms to avoid and escape anti-tumor immune responses, including suppression, inactivation, and exclusion of potential cytotoxic T cells, processes which collaborate with cells in the tumor microenvironment (TME). A better understanding of these barriers has led to a multitude of new immunomodulatory targets to be developed, some to be used in combination with e.g., checkpoint blockade or CAR T cells. On the other hand, dominant barriers to immunotherapy can be different among patients with the same cancer type, and thus there is a need for personalized approaches to immunotherapy, so that the appropriate targets are used. Here we develop a novel organotypic culture devices to maintain ex vivo cultures of primary tumors and an immune component (tumor-draining lymph nodes or circulating leukocytes), on a platform that enables precise control over spatial, molecular, cellular, and mechanical characteristics and that is relatively high-throughput to allow screening or large numbers of experimental variables. In preliminary data, we show that these devices mirror key features of in vivo responses to immunotherapy, such as improved tumor cell killing and increased markers of immunotoxicity (possible adverse events) in response to cytokine immunotherapy. We propose that these devices can be used both to screen for ideal immunotherapy combinations as well as to probe the basic mechanisms underlying the deficiencies in the anti-tumor immune response for tumors exhibiting varying levels of immune infiltration, neoantigen load, and baseline lymphatic densities. In this way, we can begin to build a stratification map that aligns key morphological features of individual tumors to treatment regimes that are most likely to lead to efficacy and tumor regression. Using a combination of both murine mouse models and primary patient-derived biospecimens, we will take advantage of the level of control afforded by our novel organotypic devices to mechanistically interrogate individual immune cell subsets and signaling axes, towards understanding their roles in influencing the course and outcomes of anti-tumor immune responses.