Autoimmunity in Cardiopulmonary Toxicities from Radiotherapy and Immunotherapy

摘要 胸部放射治疗和PD-1阻断免疫治疗后的心肺毒性是主要的 对生活质量和生存的影响。因此，有一种尚未得到满足的需求，即进行早期诊断测试和 对致命毒物的干预。我们的建议旨在了解免疫调节机制 潜在的危及生命的心肺毒性。我们建议研究这两种小鼠模型的毒性。 和病人通过分析血液样本。为了实现我们的目标，我们组建了一个协作团队 跨三个机构和一组专家顾问，以确定生物相关性和 治疗靶点，以改善这些自身免疫毒性。我们之前的研究表明， 同时暴露于放射治疗和PD-1抑制的小鼠，我们发现这依赖于两者 细胞因子IL-17A和B细胞。由于Th17/IL-17A和体液免疫都与自身免疫有关 疾病，我们假设毒性是由PD-1引起的未经检查的适应性Th17反应引起的 阻断，结合抗心和肺组织的自身抗体，由促炎症的B- 淋巴细胞。我们将使用小鼠模型和药物抑制剂来剖析潜在的 自身免疫机制及Th17和B细胞应答关键成分的前瞻性检测 采集接受放射治疗和免疫治疗的肺癌患者的血液样本。 目的1：确定IL-17A/Th17反应是否介导毒性反应。我们假设两者都是 先天免疫和获得性免疫通过IL-17A参与毒性作用。我们将产生KO小鼠 不能通过中性粒细胞或CD4T细胞产生IL-17A。我们预计毒副作用是 在这些小鼠模型中，当Th17/IL-17A被钝化时，这种作用减弱。以确定IL-17A/Th17是否可以 作为毒性的预测生物标志物，我们将检测血清中Th17/IL-17A的动态变化 我们病人的样本。目的2：确定体液反应在调节 毒物。我们假设促炎的Tbet+B淋巴细胞驱动自身抗体的产生 导致了毒副作用。我们将使用Tbetflx/Flox CD19cre小鼠作为我们的模型，在该模型中，成熟的Tbet+B细胞 缺席。这一方法将通过使用抗CD20或通过药物消耗B细胞来补充 用IVIg中和野生型小鼠的自身抗体。我们预计这些药物的毒性会减弱。 模特们。此外，我们将测试Th17/IL-17A的药物抑制剂是否会减少Tbet+B细胞和 自身抗体。最后，血液中上升的自身抗体将在小鼠和患者身上被捕获作为替代 对毒物的影响。我们对临床前模型的基本机制的研究，结合对患者的分析 样本，将导致早期发现的新诊断方法和改善严重心肺疾病的治疗 毒物。

Summary Cardiopulmonary toxicities following thoracic radiotherapy and PD-1 blocking immunotherapy have a major impact on quality of life and survival. Therefore, there is an unmet need to have early diagnostic test and intervention for fatal toxicities. Our proposal aims to understand immune mechanisms that modulate potentially life-threatening cardiopulmonary toxicities. We propose to study the toxicities in both mouse models and patients by analyzing blood samples. To achieve our goals, we have assembled a collaborative team across three institutions with a group of expert consultants in order to identify biological correlates and therapeutic targets to ameliorate these autoimmune toxicities. Our prior studies showed excessive mortality in mice simultaneously exposed to radiotherapy and PD-1 inhibition, which we show is dependent on both the cytokine IL-17A and the B-cell. Since both Th17/IL-17A and humoral immunity are implicated in autoimmune diseases, we hypothesize that toxicities result from the unchecked adaptive Th17 response due to PD-1 blockade, combined with autoantibodies against heart and lung tissues generated by the pro-inflammatory B- lymphocytes. We will employ mouse models and pharmacological inhibitors to dissect the underlying autoimmune mechanisms and measure key components of Th17 and B-cell response in prospectively collected blood samples from lung cancer patients undergoing combine radiotherapy and immunotherapy. Aim 1: To determine whether IL-17A/Th17 responses mediate the toxicities. We hypothesize that both innate and adaptive immunity contributes to the toxicities through the link of IL-17A. We will generate KO mice unable to produce IL-17A through either neutrophils or CD4 T cells. We expect that the toxicities are attenuated when Th17/IL-17A are blunted in these mouse models. To determine whether IL-17A/Th17 can be used as predictive biomarkers for the toxicities, we will examine dynamic changes of Th17/IL-17A in serum samples from our patients. Aim 2: To determine the role of humoral response in mediating the toxicities. We hypothesize that pro-inflammatory Tbet+ B-lymphocytes drive autoantibody production which results in the toxicities. We will use Tbetflox/flox CD19cre mice as our model, in which mature Tbet+ B cells are absent. This approach will be complemented by pharmacological depletion of B cells using anti-CD20 or by neutralizing autoantibodies with IVIg in wild-type mice. We expect that the toxicities are attenuated in these models. Furthermore, we will test whether pharmacological inhibitors of Th17/IL-17A reduce Tbet+ B cells and autoantibodies. Finally, the rise of autoantibodies in blood will be captured in mice and patients as a surrogate for the toxicities. Our study of basic mechanisms in preclinical models, combined with analysis of patient samples, will lead to novel diagnostics for early detection and improved therapies for severe cardiopulmonary toxicities.