Project 3: The AMPK Autophagy Pathway as a Metabolic Liability in Pancratic Ductal Adenocarcinoma

项目 3：AMPK 自噬途径作为胰腺导管腺癌的代谢负担

• 批准号: 10629065
• 负责人: Reuben Shaw
• 金额: $ 46.68万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-06 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629065
• 关键词: Address; Adenocarcinoma; Affect; Alanine; Attenuated; Automobile Driving; Autophagocytosis; Autophagosome; Biochemical; Biological Availability; Cell Compartmentation; Cell Nucleus; Cell Survival; Cell physiology; Cells; Characteristics; Chemoresistance; Chloroquine; Clinic; Clinical; Collaborations; DNA Damage; Desmoplastic; Development; Disease; Disease Progression; Duct (organ) structure; Enzymes; Epigenetic Process; Epithelial Cells; Epithelium; FRAP1 gene; Fibroblasts; Gene Expression; Genetic; Genetic Transcription; Genetically Engineered Mouse; Growth; HDAC3 gene; Histone Deacetylase; Human; KPC model; Knowledge; Lesion; MEK inhibition; MEKs; Malignant Neoplasms; Malignant neoplasm of pancreas; Measures; Mediating; Mediator; Metabolic; Metabolic stress; Metabolism; Modality; Mus; Mutation; Myelogenous; Normal Cell; Nutrient; Nutrient availability; Organelles; Organoids; Pancreas Transplantation; Pancreatic Ductal Adenocarcinoma; Paracrine Communication; Pathway interactions; Performance; Pharmaceutical Preparations; Phenotype; Phosphorylation; Phosphotransferases; Population; Proteins; Recycling; Regulation; Reporter; Reporting; Research; Resistance; Role; Signal Transduction; Source; Stress; Stromal Cells; Therapeutic; Toxic effect; Translating; Up-Regulation; Vascularization; Work; Xenograft procedure; antagonist; checkpoint inhibition; chemotherapy; defined contribution; effective therapy; efficacy evaluation; epigenetic regulation; experimental study; in vivo; inhibition of autophagy; inhibitor; kinase inhibitor; mTOR inhibition; mouse model; neoplastic cell; novel; novel strategies; pancreatic ductal adenocarcinoma model; pancreatic neoplasm; pharmacologic; programs; response; synergism; targeted treatment; therapeutic target; therapy resistant; tool; transcriptional reprogramming; treatment response; treatment strategy; tumor; tumor growth; tumor microenvironment; tumor progression

PROJECT SUMMARY – Project 3: Autophagy Pancreatic ductal adenocarcinoma (PDA) is one of the deadliest forms of cancer with few effective therapies. The poor performance of current treatments is partly due to metabolic adaptations in both the tumor and stromal compartments, such as the recycling of proteins and organelles through increased autophagy. As a hallmark of PDA, autophagy provides a key source of nutrients in the restrictive tumor microenvironment (TME). Accumulating evidence also implicates the autophagy program as a critical mediator of resistance to numerous therapeutics, including chemotherapy, MEK inhibitors, and immune checkpoint inhibition. Foundational research from the Shaw group decoded key biochemical steps involved in the initiation of autophagy, including upstream regulation of AMPK and its downstream activation of ULK1 and ULK2, the kinases that drive autophagosome formation. More recent work from the Shaw group has also revealed that AMPK can block the translocation of Class II HDACs to the nucleus. Despites these advances, the specific roles of AMPK in autophagy control and epigenetic regulation have never been investigated in pancreatic cancer. Moreover, while autophagy has emerged as an attractive therapeutic target in pancreatic cancer, efforts to translate this to the clinic have been hindered by a lack of autophagy-specific inhibitors, with only broad lysosomotropic agents like chloroquine available for study. To address this gap, the lab has developed novel, bioavailable inhibitors of ULK1 and ULK2, two of only three druggable enzymes specific to the autophagy pathway. These inhibitors provide critical tools with which to dissect the contributions of autophagy to PDA growth and form the basis for a new approach for overcoming therapeutic resistance in this deadly disease. Here, experiments proposed in Aim 1 will define when and where different facets of AMPK signaling and autophagy are activated during disease progression in the autochthonous mouse KPC model of PDA. In collaboration with Project 1, conditional deletions of AMPK or its downstream targets ULK1/2 and HDAC3 will be used to evaluate their contribution to metabolic adaptations driving tumor growth and epigenetic changes mediating tumor cellular functions. In addition, the contribution of AMPK pathway components in supporting PDA resistance to chemotherapeutics and targeted therapies will be dissected. In Aim 2, the roles of canonical and noncanonical autophagy in non-cell autonomous support of pancreatic tumor growth will be delineated by comparing stromal deletion of ULK1/2 and ATG7. In addition, experiments will dissect cell-specific requirements for autophagy within the fibroblast and myeloid compartments in supporting PDA therapeutic resistance. In Aim 3, the Shaw lab’s novel, bioavailable ULK inhibitor will be used to determine how selective inhibition of autophagy impacts PDA growth through reprogrammed tumor and stromal cell function. The potential of ULK inhibitors in rescuing autophagy-dependent therapeutic resistance to chemotherapies, MEK inhibitors, and immune checkpoint inhibition will be explored.

项目摘要 – 项目 3：自噬 胰腺导管腺癌（PDA）是最致命的癌症之一，有效的治疗方法很少。 当前治疗效果不佳的部分原因是肿瘤和基质的代谢适应 区室，例如通过增加自噬来回收蛋白质和细胞器。作为一个标志 PDA，自噬提供了限制性肿瘤微环境（TME）中的关键营养来源。 越来越多的证据也表明自噬程序是抵抗多种细菌的关键介质。 治疗方法，包括化疗、MEK 抑制剂和免疫检查点抑制。基础研究 来自Shaw小组的解码了参与自噬启动的关键生化步骤，包括上游 AMPK 的调节及其下游 ULK1 和 ULK2（驱动自噬体的激酶）的激活 形成。 Shaw 小组最近的工作还表明，AMPK 可以阻止 II 类 HDAC 至细胞核。尽管取得了这些进展，AMPK 在自噬控制和 从未在胰腺癌中研究过表观遗传调控。此外，虽然自噬 已成为胰腺癌有吸引力的治疗靶点，人们一直在努力将其转化为临床 由于缺乏自噬特异性抑制剂而受到阻碍，只有广泛的溶酶体药物如氯喹 可供学习。为了解决这一差距，该实验室开发了新型生物可利用的 ULK1 和 ULK2 抑制剂， 自噬途径特异的仅有的三种可药物酶中的两种。这些抑制剂提供了关键工具 以此剖析自噬对 PDA 生长的贡献，并为新方法奠定基础 克服这种致命疾病的治疗耐药性。在这里，目标 1 中提出的实验将定义何时 AMPK 信号传导和自噬的不同方面在疾病进展过程中被激活 PDA 的本地小鼠 KPC 模型。与项目 1 合作，有条件删除 AMPK 或其 下游目标 ULK1/2 和 HDAC3 将用于评估它们对代谢适应的贡献 驱动肿瘤生长和介导肿瘤细胞功能的表观遗传变化。此外，贡献 AMPK 通路成分支持 PDA 对化疗和靶向治疗的耐药性 解剖的。在目标 2 中，规范和非规范自噬在非细胞自主支持中的作用 通过比较 ULK1/2 和 ATG7 的基质缺失来描述胰腺肿瘤的生长。此外， 实验将剖析成纤维细胞和骨髓细胞内自噬的细胞特异性要求 支持 PDA 治疗耐药。在目标 3 中，将使用 Shaw 实验室的新型生物可利用 ULK 抑制剂 确定选择性抑制自噬如何通过重编程肿瘤影响 PDA 生长， 基质细胞功能。 ULK 抑制剂在挽救自噬依赖性治疗耐药方面的潜力 将探索化疗、MEK 抑制剂和免疫检查点抑制。