Reprogramming Metabolic Networks in the Tumor Microenvironment

项目摘要 胰腺导管腺癌（PDA）是一种严重的恶性肿瘤，迫切需要更好的治疗方法.的 胰腺肿瘤微环境是复杂的，基质和免疫细胞诱导炎症， 结缔组织增生反应，阻碍适当的血管形成，导致营养和氧气不足 交付.因此，PDA细胞必须重新连接细胞代谢以维持其生物合成和能量 要求.间质细胞和免疫细胞不仅改变了肿瘤的结构， 癌症代谢，抑制抗肿瘤免疫应答，并减弱治疗效果。事先 确定了癌细胞和癌症相关的细胞之间代谢串扰的新途径， 成纤维细胞（CAFs），我现在建议扩展这一范例，以发现免疫和神经 系统支持肿瘤代谢，以确定新的治疗机会。 肿瘤相关巨噬细胞（TAM）构成胰腺肿瘤中的大部分免疫区室 并支持恶性进展和治疗抗性。根据这一点，我们集团以前 证明TAMs释放嘧啶脱氧胞苷（dC），基于其分子相似性， 能够抑制一线化疗药物吉西他滨的细胞毒活性。由于氨基酸 天冬氨酸（Asp）是嘧啶生产所必需的，而Asp是由谷氨酸草酰乙酸产生的。 我建立了巨噬细胞特异性的Got 1或Got 2敲除模型，以研究 TAM代谢中的GOTs。在目标1a中，我将评估TAM中Asp合成和dC释放如何影响 使用同基因肿瘤模型研究PDA对吉西他滨的敏感性。我的初步数据还表明， 活性和dC调节T细胞功能。因此，在目标1b中，我将确定Got 1或Got 2的损失在 TAM影响肿瘤浸润性T细胞和肿瘤对基于免疫的治疗的反应。 在K 00阶段，我将应用研究生院在癌症生物学和代谢方面获得的知识 一个独立的博士后项目胰腺肿瘤高度神经支配和神经消融损害 肿瘤消退神经元支持癌细胞的代谢基础需要进一步研究。 调查因此，作为一名博士后研究员，我的目标是了解失调的PDA代谢是如何 影响神经元的募集和功能以及神经元中的代谢途径，这些途径对促 肿瘤信号我计划使用PDA的遗传小鼠模型，体内同位素示踪，基因组CRISPR文库， 和离体神经元-PDA共培养模型来回答这些问题。最后，除了拟议的 学习，该培训计划包括对职业发展，指导，网络， 科学交流，为我成功过渡到博士后奖学金和我的职业生涯做好准备， 研究癌症代谢的独立研究员

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDA) is a deadly disease and better therapies are urgently needed. The pancreatic tumor microenvironment is complex, and stromal and immune cells induce an inflamed, desmoplastic response that hinders proper blood vessel formation, resulting in poor nutrient and oxygen delivery. Therefore, PDA cells must rewire cellular metabolism to maintain their biosynthetic and energetic requirements. The stromal and immune cells do not simply change tumor architecture, they also support cancer metabolism, suppress anti-tumor immune responses, and blunt treatment efficacy. Having previously identified a new pathway of metabolic crosstalk between cancer cells and cancer-associated fibroblasts (CAFs), I now propose to extend this paradigm to discover how the immune and nervous systems support tumor metabolism to identify new therapeutic opportunities. Tumor-associated macrophages (TAMs) comprise the bulk of the immune compartment in a pancreatic tumor and support malignant progression and therapeutic resistance. In accordance with this, our group previously demonstrated that TAMs release the pyrimidine deoxycytidine (dC), which, based on its molecular similarity, is able to inhibit the cytotoxic activity of the frontline chemotherapeutic gemcitabine. Since the amino acid aspartate (Asp) is required for pyrimidine production, and Asp is produced by the glutamate oxaloacetate transaminases (GOTs), I generated macrophage-specific Got1 or Got2 knockout models to study the role of the GOTs in TAM metabolism. In Aim 1a, I will evaluate how Asp synthesis and dC release in TAMs impacts PDA sensitivity to gemcitabine using syngeneic tumor models. My preliminary data also suggests that GOT activity and dC modulate T cell function. Therefore, in Aim 1b, I will determine how loss of Got1 or Got2 in TAMs affects tumor infiltrating T cells and tumor response to immune-based therapy. During the K00 phase, I will apply knowledge gained during graduate school in cancer biology and metabolism to an independent postdoctoral project. Pancreatic tumors are highly innervated and neural ablation impairs tumor regression. The metabolic underpinnings by which neurons support cancer cells require further investigation. Therefore, my goal as a postdoctoral fellow is to understand how dysregulated PDA metabolism impacts neuronal recruitment and function as well as metabolic pathways in neurons that are critical for pro- tumor signaling. I plan to use genetic mouse models of PDA, in vivo isotope tracing, genome CRISPR libraries, and ex vivo neuron-PDA co-culture models to answer these questions. Lastly, in addition to the proposed studies, this training plan includes activities important for career development, mentorship, networking, and scientific communication to prepare me for successful transition to a postdoctoral fellowship and my career as an independent investigator studying cancer metabolism.