Metabolic reprogramming of the tumor microenvironment and therapy resistance

项目摘要/摘要 转移性卵巢癌(OvCa)是妇科癌症死亡的主要原因。尽管咄咄逼人 化疗和手术，大多数患者(80%)经历了腹内进展或复发到内脏 腹腔内的脂肪组织。15年多来，我的实验室一直专注于阐明 OvCa转移的生物学，重点是了解肿瘤微环境的放松 (TME)促进OvCa转移和化疗耐药。我们定义了多个基质的贡献 细胞类型到转移，揭示了甲基转移酶(NNMT)在正常 成纤维细胞通过代谢重塑转化为癌症相关的成纤维细胞。此外，我们还回答了 几十年来一直存在的问题是，为什么腹部转移的肿瘤有转移到 发现脂肪因子吸引癌细胞进入脂肪组织，脂肪细胞提供长链。 脂肪酸通过β氧化为癌细胞提供能量。然而，根本问题仍然存在。 关于OvCa进展中的代谢过程。卵巢转移瘤与原发肿瘤在代谢上有何不同 肿瘤？哪些燃料/代谢物在化疗后发生了变化，它们对化疗有何贡献？ 抵抗？鉴于免疫疗法对几种上皮性肿瘤是有效的，更令人费解和 及时的问题是为什么检查点抑制剂在Ovca中无效。我的假设是癌症与 脂肪细胞通过脂质驱动参与治疗抵抗和免疫效应细胞衰竭 TME的新陈代谢重新编程。我们已经采用了一些方法来进行体内代谢通量分析 OvCa患者，通过注入标记的代谢物(非放射性13C-葡萄糖，醋酸盐)，并正在努力 方法用成像质谱仪优化肿瘤组织的隔室拆分代谢组学。 这些数据将使我们能够确定癌症、免疫和间质细胞在治疗前后的代谢变化。 新辅助化疗。这些研究产生的假设将得到广泛的检验。 使用原始器官类型3D培养和小鼠模型的实验方法。我们的实验方法 将跨越功能细胞分析(研究黏附、迁移和侵袭)、共聚焦成像、生化 活性分析，以及新设计的方法来测试自然杀伤细胞、T细胞和 体外和体内的巨噬细胞。对肿瘤器官代谢变化的特定洞察力将 被用来开发高吞吐量的筛选活动。这些应该会发现小分子抑制剂 这可以通过建立和结构化的临床测试过程来优化。我们相信， 通过靶向肿瘤器官中确定的代谢过程，我们可以极大地加强抗肿瘤治疗 OvCa的反应，有可能阻止这种致命疾病的不可阻挡的发展特征。

PROJECT SUMMARY/ABSTRACT Metastatic ovarian cancer (OvCa) is the leading cause of death from gynecologic cancer. Despite aggressive chemotherapy and surgery, most patients (80%) experience intraabdominal progression or recurrence to visceral adipose tissue in the abdominal cavity. For more than 15 years, my laboratory has concentrated on elucidating the biology of OvCa metastasis, focusing on understanding how deregulation of the tumor microenvironment (TME) promotes OvCa metastasis and chemotherapy resistance. We defined the contribution of multiple stromal cell types to metastasis, revealing a critical role for a methyltransferase (NNMT) in the reprogramming of normal fibroblasts into cancer-associated fibroblasts through metabolic remodeling. Additionally, we answered the decades-old question of why abdominally metastasizing tumors have a propensity to metastasize to the omentum, finding that adipokines attract cancer cells to adipose tissue, and that adipocytes provide long-chain fatty acids to cancer cells for energy production through β-oxidation. However, fundamental questions remain about metabolic processes in OvCa progression. How are OvCa metastases metabolically different from primary tumors? Which fuels/metabolites are altered after chemotherapy, and how do they contribute to chemotherapy resistance? Given that immunotherapies are effective in several epithelial tumors, one of the more puzzling and timely questions is why checkpoint inhibitors are ineffective in OvCa. My hypothesis is that cancer associated adipocytes contribute to therapy resistance and immune effector cell exhaustion through the lipid-driven metabolic reprogramming of the TME. We have adapted methods to perform in vivo metabolic flux analysis in OvCa patients, by infusing labeled metabolites (non-radioactive 13C-glucose, acetate) and are working on methods to optimize compartment resolved metabolomics on tumor tissue using imaging mass spectrometry. These data will allow us to define metabolic changes in cancer, immune, and stromal cells before and after neoadjuvant chemotherapy. The hypotheses generated by these studies will be tested with wide-ranging experimental approaches using primary organotypic 3D cultures and mouse models. Our experimental approach will span functional cellular assays (to study adhesion, migration, and invasion), confocal imaging, biochemical activity assays, and newly devised methods to test the functionality of natural killer cells, T-cells, and macrophages in vitro and in vivo. Compartment-specific insights into metabolic changes in the tumor organ will be employed to develop high-throughout screening campaigns. These should discover small molecule inhibitors that can be optimized through an established and structured process towards clinical testing. We believe that, by targeting metabolic processes identified in the tumor organ, we can greatly enhance anti-tumor therapy response in OvCa, potentially halting the inexorable progression characteristic of this deadly disease.