Pro-tumor functions of neutrophils in squamous cell lung cancer

项目摘要 鳞状细胞肺癌是非小细胞肺癌的一种亚型，是第二常见的类型 肺癌的症状也被称为肺鳞状细胞癌（SCC），SCC是造成约85，000例新发 肺癌的病例每年和约50，000例死亡仅在美国。SCC不同于其他 NSCLC的主要亚型（腺癌）在其组织学、生物标志物表达、基因组改变方面， 免疫微环境和对治疗的反应。SCC的治疗选择有限，仅限于 化疗和最近的免疫疗法，其仅在一部分患者中有效。一个主要的未满足 治疗SCC的需要是确定新的治疗靶点和治疗策略， 这种疾病。SCC具有独特的组织病理学特征和复杂的微环境， 基质、角质化和免疫细胞。具体地，肿瘤相关中性粒细胞（TAN）富含 小鼠和人类SCC和中性粒细胞丰度与预后不良和对 免疫疗法关于TAN如何影响SCC肿瘤进展和治疗进展的了解相对较少。 反应众所周知，在缺乏三维结构的细胞培养中模拟SCC非常困难 完整的免疫系统和脉管系统。基因工程小鼠模型（GEMM）提供了 一个复杂的生命系统，可以高度概括人类的疾病。我们的实验室开创了 开发第一个SOX 2驱动的SCC GEMM。我们最近开发了一种快速的新GEMM， Sox 2/Lkb 1/Nkx 2 -1（SNL）的改变，其中SCC肿瘤在以下方面与人类疾病高度相似： 组织病理学、遗传学和免疫微环境。这种新的SNL模型代表了一种独特的免疫- 是研究TAN如何影响肿瘤进展和治疗反应的有力工具。的目标 本研究旨在阐明TANs在肺鳞癌发生、发展和对化疗反应中的作用。 疗法我们假设TAN通过增加活性氧（ROS）来促进鳞状细胞的命运， 其与肿瘤转分化有关，并通过促进中性粒细胞胞外陷阱（NETosis）。到 为了验证这些假设，我们将：1）确定TAN改变肿瘤细胞命运的机制。2)确定 TAN是否通过ROS和/或NETosis促进鳞状肺癌进展。3)确定是否 嗜中性粒细胞耗竭增强了对化疗和/或免疫治疗的反应。这种做法是创新的 因为我们将采用：1）我们的新型免疫能力SCC GEMM，它概括了 人类疾病，2）临床试验中的中性粒细胞耗竭策略，以及3）最新技术 包括单细胞RNA测序和NETosis的天然癌症相关抑制剂。本研究是 这是因为TAN在SCC中的功能几乎完全未被探索，TAN可能代表了一种新的 这一难治性疾病的治疗目标。

Project Summary Squamous cell lung cancer, a subtype of non-small cell lung cancer (NSCLC), is the second most common type of lung cancer. Also known as squamous cell carcinoma (SCC) of the lung, SCC is responsible for ~85,000 new cases of lung cancer each year and ~50,000 deaths in the United States alone. SCC is distinct from the other major subtype of NSCLC (adenocarcinoma) in terms of its histology, biomarker expression, genomic alterations, immune microenvironment and response to therapy. SCC has limited therapeutic options that are confined to chemotherapy and more recently, immunotherapy, which is only effective in a subset of patients. A major unmet need for the treatment of SCC is the identification of new therapeutic targets and treatment strategies to combat this disease. SCC has a unique histopathology and a complex microenvironment that is rich with desmoplastic stroma, keratinization and immune cells. Specifically, tumor-associated neutrophils (TANs) are enriched in mouse and human SCC, and neutrophil abundance correlates with poor prognosis and poor response to immunotherapy. Relatively little is known about how TANs impact SCC tumor progression and therapeutic response. It is notoriously difficult to model SCC in cell culture, which lacks the three-dimensional architecture of tumors, an intact immune system, and vasculature. Genetically-engineered mouse models (GEMMs) provide a complex living system that can highly recapitulate the human disease. Our laboratory pioneered the development of the first SOX2-driven SCC GEMM. We recently developed a rapid new GEMM based on alterations in Sox2/Lkb1/Nkx2-1 (SNL) where SCC tumors highly resemble the human disease in terms of histopathology, genetics, and immune microenvironment. This new SNL model represents a unique immune- competent tool to investigate how TANs shape tumor progression and therapeutic response. The objective of this study is to elucidate the role of TANs in squamous lung cancer development, progression and response to therapy. We hypothesize that TANs promote squamous cell fate by increasing reactive oxygen species (ROS), which are implicated in tumor transdifferentiation, and by promoting neutrophil extracellular traps (NETosis). To test these hypotheses, we will: 1) Determine the mechanism by which TANs alter tumor cell fate. 2) Determine whether TANs promote squamous lung cancer progression via ROS and/or NETosis. 3) Determine whether neutrophil depletion enhances response to chemotherapy and/or immunotherapy. This approach is innovative because we will employ: 1) our novel immune-competent SCC GEMM that recapitulates key features of the human disease, 2) neutrophil depletion strategies that are in clinical trials, and 3) state-of-the-art technologies including single cell RNA-sequencing and natural cancer-associated inhibitors of NETosis. This research is significant because TAN function in SCC is almost completely unexplored and TANs may represent a new therapeutic target for this intractable disease.