Neutrophil Extracellular Traps and the Intersection of the Immune and Biophysical Microenvironments

项目概述：虽然近年来免疫疗法彻底改变了癌症治疗，但主要的 挑战依然存在。转移性癌症仍然难以治疗，导致90%的癌症死亡，以及 只有20%-40%的患者对免疫治疗有反应，不同癌症的反应程度不同。在……里面 实体瘤、淋巴管活化或淋巴管生成与转移和改善有关。 对免疫治疗的反应，以及最近，中性粒细胞的激活也在 这两个过程。中性粒细胞促进癌症进展的一个重要机制是通过 中性粒细胞胞外陷阱(Net)的形成。网络是解压缩的DNA的网状网络 点缀着组蛋白和颗粒蛋白，它们由中性粒细胞释放，以捕获和杀死 病原体，或对肿瘤分泌的细胞因子的反应。这些蛋白质中有许多是蛋白水解酶，它们具有 重塑细胞外基质(ECM)的潜力。在这个提案中，我们将探索这部小说 假设淋巴管和中性粒细胞合作，以改变 细胞外基质与宿主免疫反应。我们将使用免疫荧光染色和第二个 从4T1和B16小鼠肿瘤模型到B16肿瘤模型的肿瘤、肺和淋巴结的谐波成像 描述肿瘤转移中NETsis、淋巴管生成和细胞外基质之间的相互作用。我们还将 使用特定于肺的可诱导的淋巴管生成模型，以确定 转移的发生与转移前的生态位或肿瘤微环境有关。我们将使用 生理性人工ECM和体外肿瘤-淋巴结微环境三维灌流模型的建立 以确定肿瘤微环境的生物物理特性如何影响NETsis和T细胞 细胞毒性，以及蚊虫感染如何影响肿瘤进展和对免疫治疗的反应。由此产生的结果 这项工作可以帮助我们更好地预测哪些患者可能对免疫治疗有反应，并导致新的 使免疫治疗在更多患者中更有效的治疗策略。它还将增加我们的 了解微环境的生物物理性质如何影响免疫细胞功能 这可能是许多疾病过程中的一个重要因素。

Project Summary: While immunotherapy has revolutionized cancer treatment in recent years, major challenges still remain. Metastatic cancer remains difficult to treat, resulting in 90% of cancer deaths, and only 20-40% of patients respond to immunotherapy, with varying levels of response across cancers. In solid tumors, lymphatic activation, or lymphangiogenesis, is associated with both metastasis and improved response to immunotherapy, and more recently, neutrophil activation has also emerged as playing a key role in both of these processes. An important mechanism by which neutrophils promote cancer progression is through the formation of neutrophil extracellular traps (NETs). NETs are a mesh network of decondensed DNA dotted with histones and granular proteins, and they are released by neutrophils in order to trap and kill pathogens, or in response to tumor-secreted cytokines. Many of these proteins are proteases, which have the potential to remodel the extracellular matrix (ECM). In this proposal, we will explore the novel hypothesis that lymphatics and neutrophils collaborate in driving pro-metastatic programs in ways that alter the extracellular matrix (ECM) and host immune response. We will use immunofluorescence staining and second harmonic generation imaging of tumors, lungs, and lymph nodes from 4T1 and B16 murine models of cancer to characterize the cross-talk between NETosis, lymphangiogenesis and ECM in cancer metastasis. We will also use an inducible model of lymphangiogenesis that is specific to the lung in order to determine if the effect of NETosis on metastasis is specific to the pre-metastatic niche or the tumor microenvironment. We will use physiological artificial ECM and 3D perfusion models of the tumor-lymph node microenvironment with ex vivo tissues to determine how the biophysical properties of the tumor microenvironment affect NETosis and T cell cytotoxicity, and how NETosis affects tumor progression and response to immunotherapy. Results from this work could help us better predict which patients are likely to respond to immunotherapy and lead to new therapeutic strategies that make immunotherapy more effective in more patients. It will also increase our understanding of how the biophysical properties of the microenvironment affect immune cell function in general, which is likely an important factor in many disease processes.