Research Project Cervical Cancer

项目1概要（CERVIX） 局部晚期宫颈癌的标准治疗（SOC），盆腔放疗（RT）联合顺铂， 与30 - 50%的失败率相关，并且没有治愈复发性疾病的方法。大多数宫颈癌是 人乳头瘤病毒（HPV）感染引起的，HPV癌基因的持续表达支持 一种由巨噬细胞介导的慢性基质炎症状态。临床前研究表明，肿瘤 相关巨噬细胞（TAM）是RT后肿瘤微环境（TME）中最丰富的细胞类型 并表现出抑制抗肿瘤免疫发展的"M2样"表型。在我们之前的工作中， 我们证明了宫颈肿瘤18F-氟脱氧葡萄糖（FDG）摄取在治疗前正电子发射 断层扫描（PET）与宫颈TME中具有"M2样"表型的TAM增加相关， 用TAM培养宫颈肿瘤细胞增加肿瘤细胞葡萄糖摄取。我们认为TAM和肿瘤 TME中的细胞共同进化并相互适应其葡萄糖和ROS代谢以优化肿瘤生长。在 在这个提议中，我们将检验SOC CRT诱导ROS介导的"代谢开关"的假设， 在每种细胞类型中的基因转录和代谢，提供限制这种代谢的治疗机会， 共依赖性，并通过促进RT诱导的抗肿瘤免疫来改善SOC CRT后的结局。这里我们 将使用最先进的单细胞和人类肿瘤的空间分辨谱的组合， 的状态，以确定SOC CRT诱导的肿瘤细胞代谢变化的影响， TAM对CRT敏感性的影响，并进行机制临床前研究，以确定这如何影响 抗肿瘤免疫的发展。在目标1中，我们将确定SOC CRT的纵向影响， 代谢和炎症信号。我们的初步数据支持针对CRT 诱导肿瘤细胞中SOD 2和NRF 2的上调。我们将研究CRT相关的ROS变化， 使用肿瘤类器官和新型小鼠模型进行线粒体代谢。在目标2中，我们将确定 SOC CRT对宫颈TME中TAM和T细胞表型和功能的影响。我们将研究TAM的作用 在小鼠模型中使用新型成像方法、抗体耗竭和CCR2抑制剂进行靶向。在目标3 我们将整合PET成像特征的纵向变化与CRT相关的基因表达变化， 以改善患者分类器和反应预测。在这些研究中，我们将使用FDG和64 Cu- DOTA-ECLi，用于CCR 2 + TAM的新型PET示踪剂。最后，我们将与来自培训中心的学员合作， METEORITE，优先考虑我们人类数据中的新线索。本学员将探索免疫系统的贡献 检查点阻滞（ICB）添加至SOC CRT时。众所周知，ICB会改变免疫细胞的代谢， 我们将综合我们的数据和方法，以实现我们的最终目标， 免疫代谢转化为新的放射治疗策略。这些实验将优化一种新的治疗方法 联合重编程TAM代谢作为改善SOC CRT + ICB后结局的新方法。

PROJECT 1 SUMMARY (CERVIX) The standard of care (SOC) for locally advanced cervical cancer, pelvic radiation (RT) with concurrent cisplatin, is associated with a 30-50% failure rate, and there is no cure for recurrent disease. Most cervical cancers are caused by infection with Human Papilloma Virus (HPV), and persistent expression of HPV oncogenes supports a state of chronic stromal inflammation mediated by macrophages. Preclinical studies suggest that tumor associated macrophages (TAMs) are the most abundant cell type in the tumor microenvironment (TME) after RT and exhibit “M2-like” phenotypes that suppress the development of anti-tumor immunity. In our previous work, we demonstrated that cervical tumor 18F-Fluoro-deoxy-glucose (FDG) uptake on pretreatment positron emission tomography (PET) is associated with increased TAMs with “M2-like” phenotypes in the cervix TME, and that co- culture of cervix tumor cells with TAMs increases tumor cell glucose uptake. We propose that TAMs and tumor cells in the TME co-evolve and mutually adapt their glucose and ROS metabolism to optimize tumor growth. In this proposal, we will test the hypothesis that SOC CRT induces ROS-mediated “metabolic switches” that alter gene transcription and metabolism in each cell type, providing a therapeutic opportunity to limit this metabolic co-dependency and improve outcomes after SOC CRT by promoting RT induced anti-tumor immunity. Here we will use a combination of state of the art single cell and spatially resolved profiling of human tumors together with state of the are imaging to determine the impact of SOC CRT induced changes in metabolism in tumor cells and TAMs on CRT sensitivity, and perform mechanistic preclinical studies to determine how this impacts the development of anti-tumor immunity. In Aim 1 we will determine the longitudinal impact of SOC CRT on metabolism and inflammatory signaling in cervix tumor cells. Our preliminary data supports targeting CRT induced upregulation of SOD2 and NRF2 in tumor cells. We will study CRT associated changes in ROS and mitochondrial metabolism using tumor organoids and novel mouse models. In Aim 2 we will determine the impact of SOC CRT on TAM and T cell phenotype and function in the cervix TME. We will study the effect of TAM targeting using novel imaging approaches, antibody depletion and CCR2 inhibitors in mouse models. In Aim 3 we will integrate longitudinal changes in PET imaging features with CRT associated changes in gene expression to improve patient classifiers and response prediction. In these studies, we will use FDG in addition to 64Cu- DOTA-ECLi, a novel PET tracer for CCR2+ TAMs. Finally, we will work with a trainee from our Training Core, METEORITE, to prioritize new leads from our human data. This trainee will explore the contribution of immune checkpoint blockade (ICB) when added to SOC CRT. ICB is well known to alter metabolism in immune cells, and we will synthesize our data and approaches to achieve our ultimate goal which is to incorporate immunometabolism into novel radiosentization strategies. These experiments will optimize a new treatment combination reprogramming TAM metabolism as a novel means to improve outcomes after SOC CRT + ICB.