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

项目概要/摘要 免疫疗法在为许多癌症和转移性疾病提供治疗方面有着巨大的希望， 使一部分患者受益。肿瘤仍然可以通过多种机制来避免和逃避抗肿瘤治疗。 免疫应答，包括潜在的细胞毒性T细胞的抑制、失活和排除， 它们与肿瘤微环境（TME）中的细胞合作。更好地理解这些障碍， 导致许多新的免疫调节靶点被开发，其中一些将与 例如，在一个实施例中，检查点阻断或CAR T细胞。另一方面，免疫治疗的主要障碍可能是 在具有相同癌症类型的患者之间是不同的，并且因此需要个性化的方法来 免疫疗法，以便使用适当的目标。在这里，我们开发了一种新的器官型培养装置 为了维持原发性肿瘤和免疫组分（肿瘤引流淋巴结或 循环白细胞），在一个平台上，能够精确控制空间，分子，细胞，和 机械特性，并且其具有相对高的通量以允许筛选或大量的 实验变量在初步数据中，我们表明这些设备反映了体内 对免疫疗法的反应，如改善的肿瘤细胞杀伤和增加的免疫毒性标志物 （可能的不良事件）。我们建议这些设备可以用于 既筛选理想的免疫治疗组合，也探索免疫治疗的基本机制。 对表现出不同免疫浸润水平的肿瘤的抗肿瘤免疫应答的缺陷， 新抗原负荷和基线淋巴密度。这样，我们就可以开始建立一个分层图， 将个体肿瘤的关键形态学特征与最有可能导致 疗效和肿瘤消退。使用鼠模型和原发性患者来源的 生物标本，我们将利用我们的新型器官型设备提供的控制水平， 机械地询问个体免疫细胞亚群和信号传导轴，以了解它们的 在影响抗肿瘤免疫应答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.