Metabolic Contributions of Individual Cellular Compartments to the Diversity of the Tumor Microenvironment in Renal Cell Carcinoma

项目总结 解除对细胞能量学的管制和避免免疫破坏被认为是癌症的标志。 激发抗肿瘤免疫现在是癌症治疗的主要目标。免疫检查点的成功 阻断(ICB)疗法展示了这种范式的巨大前景，但仍然只有一小部分 患者对这种治疗方式有持久的反应。我建议改变新陈代谢程序 肿瘤微环境(TME)可能与抗肿瘤免疫反应功能障碍有关。通过揭示这些 通过相互作用，可能会有新的机会来提高免疫疗法的疗效。我创作了一部小说 数据显示，跨越肿瘤模型的髓细胞在TME中摄取显著更多的葡萄糖，而 转化的细胞似乎正在消耗谷氨酰胺。肾细胞癌(RCC)的代谢改变 以复杂而丰富的免疫细胞渗入为特征的肿瘤。这是由我们的 小组和其他人认为这些肿瘤浸润性T细胞在代谢上受到损害，抗肿瘤作用有限 容量。尽管ICB提高了肾癌患者的存活率，但只有少数患者完全 对这些T细胞刺激剂的反应。肾癌独特的遗传学可能与此有关。 前述抑制性TME。在透明细胞肾癌中，肿瘤抑制因子von Hippel Lindau(VHL)的缺失 是肿瘤发生过程中必不可少的事件。此外，在侵袭性II型乳头状RCC的子集中，丢失 肿瘤的形成需要富马酸水合酶(FH)或三羧酸循环中的其他缺陷。这些 遗传事件是由我的导师WK Rathmell和其他人定义的。这些跨越RCC结果的遗传事件 在致癌转录因子(TF)的积累中，缺氧诱导因子1和2。使用 这些TF的积累，RCC肿瘤细胞通过减少对 三元酸循环和线粒体呼吸，同时增加细胞糖酵解。因此，RCC是 为进一步研究肿瘤细胞代谢对淋巴系和髓系细胞命运的影响而独特提出 功能。本项目将应用新的免疫活性、非免疫原性CRISPR/Cas9模型来研究 肾癌基因事件(VHL和FH缺失)对免疫渗透和激活的影响。在这些模型中，我 还将研究抑制葡萄糖和谷氨酰胺摄取对免疫细胞的不同结果 在TME中的体能和机能。这项工作将得到使用体外原代培养的研究的补充 检测遗传和化学扰动对人类的影响的人类RCC有机体模型 肿瘤常驻免疫细胞代谢。这项研究将最终阐明肿瘤的异型性。 新陈代谢。通过了解异源TME中关键细胞类型的不同代谢能力， 这项工作将提高我们支持浸润性免疫细胞抗肿瘤能力的能力。这些 数据将有助于未来的药理学策略，这些策略可以提高我们诱导具有临床意义的 在更大的患者队列中的抗肿瘤免疫反应。

PROJECT SUMMARY Deregulating cellular energetics and avoiding immune destruction are considered hallmarks of cancer. Stimulating anti-tumor immunity is now a chief goal of cancer therapy. The success of immune checkpoint blockade (ICB) therapy demonstrates the tremendous promise of this paradigm, but still only a minority of patients have durable responses with this modality of therapy. I propose that altered metabolic programs in the tumor microenvironment (TME) may be linked to dysfunctional anti-tumor immune responses. By revealing these interactions, there may be new opportunities to improve the efficacy of immunotherapy. I have generated novel data demonstrating that myeloid cells across tumor models uptake significantly more glucose in the TME, while transformed cells appear to be glutamine consuming. Renal Cell Carcinomas (RCC) are metabolically altered tumors that are characterized by a complex and abundant immune cell infiltrate. It is well established by our group and others that these tumor-infiltrating T cells are metabolically compromised and have limited antitumor capacity. Even though ICB has improved RCC patient survival, only a minority of patients have complete responses with these T cell stimulating agents. The unique genetics of RCC may contribute to this aforementioned suppressive TME. In clear cell RCCs, the loss of the tumor suppressor von Hippel Lindau (VHL) is a necessary event for tumorigenesis. Additionally, in a subset of aggressive Type II papillary RCCs, the loss of fumarate hydratase (FH) or other defects in the tricarboxylic acid cycle are required for tumor formation. These genetic events were defined by my mentor WK Rathmell and others. These genetic events across RCC results in accumulation of the oncogenic transcription factors (TFs) hypoxia inducible factor 1 and 2. With accumulation of these TFs, RCC tumor cells shift their energetic requirements by decreasing their reliance on the tricarboxylic acid cycle and mitochondrial respiration while increasing cellular glycolysis. Therefore, RCC is uniquely posed to further study the impact of tumor cell metabolism on lymphoid and myeloid cell fate and function. This project will apply novel immunocompetent, non-immunogenic CRISPR/Cas9 models to study the effect of RCC genetic events (VHL and FH loss) on immune infiltration and activation. In these models, I will also examine the differential outcomes of inhibiting glucose and glutamine uptake on immune cell fitness and function in the TME. This work will be complemented by studies that employ in vitro primary human RCC organoid models to examine the impact of genetic and chemical perturbations on human tumor resident immune cell metabolism. This study will ultimately shed light on the heterotypic nature of tumor metabolism. By understanding the divergent metabolic capacities of the key cell types in the heterogenous TME, this work will increase our capability to support anti-tumor capacity of infiltrating immune cells. These data will aid future pharmacological strategies that can increase our ability to induce clinically significant anti-tumor immune responses in larger cohorts of patients.