The Induction of Macrophage Endoplasmic Reticulum Stress by Irradiated-Tumor Derived Extracellular Vesicles Supports the Adoption of a Pro-Tumor Phenotype

项目摘要 放射治疗启动了交替刺激（M2）巨噬细胞向肿瘤的募集 微环境这些细胞表现出“促肿瘤表型”，刺激血管生成，抑制抗肿瘤活性。 增强肿瘤细胞的抗辐射能力。这限制了放射治疗的总体疗效， 实体瘤如非小细胞肺癌目前放射免疫学领域的研究主要集中在 这些操作要么抑制这些细胞的募集，要么一旦它们到达细胞膜就改变它们的行为。 肿瘤微环境然而，辐射癌细胞如何操纵巨噬细胞在很大程度上是未知的。 未知我们已经发现，照射后的肺癌细胞（IR-EVs） 诱导ER应激并引发SRA/CD 204中巨噬细胞产生“促肿瘤”细胞因子 依赖的方式。本项目的总体目标是确定IR-EV的分子机制， 诱导巨噬细胞促肿瘤细胞因子表达。首先，我们将测试IR-EV诱导的功能影响。 使用CHOP-/-C57 BL/6小鼠和可溶性ER抑制剂对巨噬细胞表型巨噬细胞ER应激 应力然后，我们将研究巨噬细胞SRA/CD 204在巨噬细胞对IR-EV的反应中的作用。 我们将使用SRA/CD 204-/-C57 BL/6小鼠以及开发的新型SRA/CD 204阻断抗体来实现这一点。 在我们的实验室里。最后，我们将比较复发性和非复发性非小细胞肺癌的循环IR-EV。 肺癌患者，以确定他们是否可以预测放疗的结果。来自经常性和非经常性 在RT之前、期间和之后分离的复发性NSCLC患者将评估其刺激肿瘤细胞增殖的能力。 体外促肿瘤巨噬细胞表型。然后，质谱分析将被用来识别一个潜在的 SRA/CD 204结合蛋白在来自患者和培养细胞的IR-EV表面上上调。的潜在 携带配体的外泌体将肺癌患者分为复发或无病生存组， 使用市售ELISA对浓缩的血清外泌体进行评价。

Project Summary Radiotherapy initiates the recruitment of alternatively stimulated (M2) macrophages to the tumor microenvironment. These cells exhibit a “pro-tumor phenotype,” stimulating angiogenesis, suppressing anti- tumor immunity, and enhancing tumor cell radioresistence. This limits the overall efficacy of radiotherapy for solid tumors such as non-small cell lung cancer. Current research in the field of radioimmunology is focused on manipulations that either inhibit the recruitment of these cells or that alter their behavior once they reach the tumor microenvironment. However, exactly how irradiated cancer cells manipulate macrophages is largely unknown. We have discovered that extracellular vesicles elaborated by irradiated lung cancer cells (IR-EVs) induce ER-stress and elicit the production of “pro-tumor” cytokines from macrophages in an SRA/CD204 dependent manner. The overall objective of this project is to define the molecular mechanisms by which IR-EVs induce macrophage pro-tumor cytokine expression. First, we will test the functional impact of IR-EV induced macrophage ER stress on macrophage phenotype using CHOP-/- C57BL/6 mice and soluble inhibitors of ER stress. Then, we will investigate the role of macrophage SRA/CD204 in the response of macrophages to IR-EVs. We will do this using SRA/CD204-/- C57BL/6 mice as well as a novel SRA/CD204 blocking antibody developed in our laboratory. Finally, we will compare the circulating IR-EVs of recurrent and non-recurrent non-small cell lung cancer patients to determine if they can predict the outcome of radiotherapy. EVs from recurrent and non- recurrent NSCLC patients isolated before, during, and after RT will be assessed for their ability to stimulate a pro-tumor macrophage phenotype in vitro. Then, mass spectrometry will be used to identify a potential SRA/CD204 binding protein upregulated on the surface of IR-EVs from patients and cultured cells. The potential of ligand bearing exosomes to segregate lung cancer patients into relapse or disease-free survival groups will be evaluated using a commercially available ELISA on concentrated serum exosomes.