Development of Mouse and Humanized Models to Study Sex Disparities in Tumor Progression and Treatment of NSCLC

项目摘要 根据美国癌症协会的数据，肺癌是男性第二大常见的癌症 和妇女除了性别特异性癌症外，肺癌和黑色素瘤的性别差异最大， 尽管这些差异背后的确切机制还没有很好地理解。在美国，男性 患肺癌的终生风险更高，而且比女性更容易患上严重的疾病。 此外，在对治疗的反应方面存在性别差异，例如男性对治疗的反应更好。 免疫疗法优于女性。有多种因素导致这种性别差异，包括生活方式 选择、性激素和免疫反应的差异。我们的初步数据显示两个小鼠肿瘤 肺癌模型，CMT-167和刘易斯肺癌（LLC），两种非小细胞肺癌模型 （NSCLC），在雌性小鼠中生长慢于雄性小鼠。这种性别差异是依赖于卵巢肿瘤， 切除卵巢的雌性小鼠的生长与雄性小鼠相当。天然免疫细胞（巨噬细胞和NK 细胞），更具体地说，NKG2D受体活性是降低女性肿瘤生长所必需的。多- 参数流式细胞术分析显示NSCLC肿瘤固有免疫的显著性别差异 细胞我们发现，这些性别差异延伸到几种化疗和抗PDL 1免疫治疗， 治疗。在初步的体内数据中，我们表明NSCLC肿瘤生长的性别差异和对化疗的反应， 化疗需要NK细胞。离体NK细胞杀伤测定显示，NSCLC细胞与 女性血清，而不是男性血清或来自卵巢切除女性的血清，刺激增强的NK细胞活性 利用分泌的促凋亡因子TRAIL。这种离体NK细胞测定提供了鉴定生物活性的方法， 分子。基于这些初步数据，我们提出了两个目标，为我们未来的研究。目标1我们将 从雌性小鼠的血清中鉴定LLC所需的来自雌性血清的生物分子， CMT-167通过TRAIL致敏NK细胞杀伤。LLC和CMT-167肿瘤细胞在女性中的致敏性 小鼠对TRAIL的影响被认为有助于观察到的肿瘤生长的性别差异， 对治疗（化疗和免疫治疗）的敏感性。对于目标2，我们将确定响应是否 使用原位CMT-167和LLC肿瘤的标准护理化疗或抗PD1免疫疗法 模特依赖于性别。如果观察到反应的性别差异，我们将描述免疫系统的特征。 在这些药物的背景下，雄性和雌性小鼠对肺肿瘤的反应。总而言之，该项目旨在 了解NSCLC进展中性别差异背后的生物学机制， 对治疗的反应，希望获得知识，为临床决策和人类治疗提供信息。 疾病

Project Summary According to the American Cancer Society, lung cancer is the second most commonly diagnosed cancer in men and women. Outside of the sex-specific cancers, lung cancer and melanoma have the greatest sex disparity, though the exact mechanisms behind these differences are not well understood. In the United States, men have a higher lifetime risk of developing lung cancer and are more likely to develop severe disease than women. Additionally, there are sex disparities in responses to treatment, such as men having better responses to immunotherapies over women. There are multiple factors contributing to this sex disparity including lifestyle choices, sex hormones, and differences in immune response. Our preliminary data shows that two mouse tumors models of lung cancer, CMT-167 and Lewis Lung Carcinoma (LLC), two models of non-small cell lung cancer (NSCLC), grow slower in female than in male mice. This sex difference is dependent on the ovary as tumors in ovariectomized female mice grow equivalent to those in male mice. Innate immune cells (macrophages and NK cells) and more specifically NKG2D receptor activity are required for reduced tumor growth in females. Multi- parameter flow cytometry analysis shows significant sex differences in NSCLC tumor resident innate immune cells. We show that these sex-disparities extend to several chemotherapy and anti-PDL1 immunotherapy treatments. In preliminary in vivo data we show that the sex-disparity in NSCLC tumor growth and the response to chemotherapy requires NK cells. Ex vivo NK cell killing assays show that preincubation of NSCLC cells with female serum, but not male serum or serum from ovariectomized females, stimulates enhanced NK cell activity utilizing the secreted pro-apoptotic factor TRAIL. This ex vivo NK cell assay provides a means to identify bioactive molecules. Based upon this preliminary data, we propose two Aims for our future studies. Aim 1 we will fractionate serum from female mice to identify a biological molecule from female serum required for LLC and CMT-167 sensitization to NK cell killing via TRAIL. The sensitization of LLC and CMT-167 tumor cells in female mice to the effects of TRAIL are proposed to contribute to the observed sex–differences in tumor growth and sensitivity to therapies (both chemotherapy and immunotherapy). For Aim 2, we will determine if the response to standard of care chemotherapies or anti-PD1 immunotherapy using orthotopic CMT-167 and LLC tumor models is dependent on sex. If a sex difference in response is observed, we will characterize the immune responses of male and female mice to lung tumors in the context of these drugs. In summary, this project aims to gain an understanding of the biological mechanisms behind the sex disparities in NSCLC progression and response to treatment, with the hopes of gaining knowledge to inform clinic decisions and treatment of the human disease.