Reprogramming Metabolic Networks in the Tumor Microenvironment

肿瘤微环境中的代谢网络重编程

• 批准号: 10807199
• 负责人: Samuel Andrew Kerk
• 金额: $ 8.26万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10807199
• 关键词: Ablation; Affect; Amino Acids; Animals; Architecture; Aspartate; Biochemical Pathway; Bioenergetics; CRISPR library; Cancer Biology; Cell physiology; Cells; Chemicals; Clinical Trials; Coculture Techniques; Communication; Complex; Coupled; Data; Deoxycytidine; Desmoplastic; Disease; Doctor of Philosophy; Effector Cell; Equilibrium; Fellowship; Fibroblasts; GOT2 gene; Genes; Genetic; Genetic Screening; Genome; Glutamates; Goals; Growth; Immune; Immune system; Immunocompetent; Immunotherapy; Impairment; Infiltration; Investigation; Isotopes; Knock-out; Knowledge; Libraries; Macrophage; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Mentorship; Metabolic; Metabolic Pathway; Metabolism; Mitochondria; Modeling; Molecular; Mus; Myelogenous; Nature; Nerve; Nervous System; Neurons; Neurotransmitters; Nutrient; Oxaloacetates; Oxidation-Reduction; Oxygen; Pancreatic Ductal Adenocarcinoma; Peripheral Nerves; Phase; Physiological; Population; Postdoctoral Fellow; Production; Proliferating; Pyrimidine; Pyrimidines; Pyruvate; Regulatory T-Lymphocyte; Repression; Research Personnel; Resistance; Role; Signal Transduction; Support System; Supporting Cell; T-Lymphocyte; Technical Expertise; Techniques; Testing; Therapeutic; Tissues; Training; Transaminases; Treatment Efficacy; Tumor-Infiltrating Lymphocytes; Tumor-associated macrophages; Vertebral column; Work; amino acid metabolism; anti-tumor immune response; blood vessel development; cancer cell; cancer infiltrating T cells; career; career development; chemotherapy; conditional knockout; cytotoxic; cytotoxicity; gemcitabine; graduate school; in vivo; in vivo Model; inhibitor; metabolic abnormality assessment; metabolomics; mitochondrial metabolism; mouse model; neural; neural network; neurotransmission; novel therapeutics; nucleobase; pancreatic cancer cells; pancreatic cancer model; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; pancreatic neoplasm; recruit; response; targeted treatment; therapy resistant; tumor; tumor ablation; tumor growth; tumor metabolism; tumor microenvironment; tumorigenesis; tumorigenic

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.

项目概要 胰腺导管腺癌（PDA）是一种致命的疾病，迫切需要更好的治疗方法。这 胰腺肿瘤微环境复杂，基质细胞和免疫细胞诱发炎症、 促纤维增生反应阻碍正常的血管形成，导致营养和氧气不足 送货。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。基质细胞和免疫细胞不仅仅改变肿瘤结构，它们还支持 癌症代谢，抑制抗肿瘤免疫反应，削弱治疗效果。之前有过 发现了癌细胞和癌症相关物质之间代谢串扰的新途径 成纤维细胞（CAF），我现在建议扩展这个范式来发现免疫和神经细胞如何 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) 构成胰腺肿瘤中免疫区室的大部分 并支持恶性进展和治疗抵抗。据此，我们组此前 证明 TAM 释放嘧啶脱氧胞苷 (dC)，根据其分子相似性，它是 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，而天冬氨酸是由谷氨酸草酰乙酸产生的 转氨酶 (GOT) 中，我生成了巨噬细胞特异性 Got1 或 Got2 敲除模型来研究转氨酶 (GOT) 的作用 TAM 代谢中的 GOT。在目标 1a 中，我将评估 TAM 中的 Asp 合成和 dC 释放如何影响 使用同基因肿瘤模型观察 PDA 对吉西他滨的敏感性。我的初步数据还表明 GOT 活性和 dC 调节 T 细胞功能。因此，在目标 1b 中，我将确定 Got1 或 Got2 的损失如何 TAM 影响肿瘤浸润 T 细胞和肿瘤对免疫治疗的反应。 在K00阶段，我将应用在研究生院获得的癌症生物学和代谢知识 独立的博士后项目。胰腺肿瘤高度神经支配，神经消融会损害 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，作为博士后，我的目标是了解 PDA 代谢失调是如何发生的 影响神经元的募集和功能以及神经元中的代谢途径，这对于亲和性至关重要 肿瘤信号传导。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，除了提议的 研究中，该培训计划包括对职业发展、指导、网络和 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者。