Targeting Stromal Influences on BCKA Addiction in PDAC Tumors

靶向基质对 PDAC 肿瘤 BCKA 成瘾的影响

基本信息

批准号：
10453984
负责人：
THEODORE S LAWRENCE
金额：
$ 58.14万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-01 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10453984
关键词：
Allografting Anabolism Branched-Chain Amino Acids Cell Line Cells Cessation of life Citric Acid Cycle Clustered Regularly Interspaced Short Palindromic Repeats Communication Complex Coupling Cytosol Data Dependence Ecosystem Epithelial Event Extracellular Matrix Extracellular Matrix Degradation Failure Fibroblasts Foundations Genes Genetic Goals Growth In Vitro Isotopes Keto Acids Knockout Mice Knowledge Lead Lipids MADH4 gene Malignant Neoplasms Malignant neoplasm of pancreas Metabolic Metabolic Pathway Methods Mitochondria Modeling Nature Neoplasm Circulating Cells Neoplasm Metastasis Nucleotides Nutrient Organoids Oxidoreductase Pancreas Pancreatic Ductal Adenocarcinoma Pathway interactions Patients Play Production Proteolysis Public Health Reaction Recurrence Regulation Role Sampling Slice Source Stromal Cells Symbiosis System Testing Time Tissues Tumor Volume addiction amino acid metabolism base branched-chain-amino-acid transaminase cancer cell cancer therapy cell growth cell transformation conditional knockout drug development efficacy testing gabapentin in vivo in vivo Model inhibitor insight loss of function new therapeutic target novel oxidation pancreatic cancer cells pancreatic ductal adenocarcinoma cell pancreatic ductal adenocarcinoma model tool traditional therapy tumor tumor metabolism tumor microenvironment

项目摘要

Failure of traditional therapy in pancreatic ductal adenocarcinoma (PDAC) is due to our limited understanding of how the tumor microenvironment (TME) can facilitate the rapid progression or recurrence of PDAC. In PDAC, the stromal cells identified as cancer associated fibroblasts (CAFs) account for up to 90% of tumor volume. Recent studies have revealed the significance of branched chain amino acids (BCAAs) in cancer. However, the role of stromal cells in support of BCAA metabolism in tumors is still poorly understood. Understanding cancer-stromal ecosystem requires insight into the intersection of cancer- associated transformations in the stroma with reprogramming of their BCAA metabolism. SMAD4 deletion is a frequent event in PDAC. However, the metabolic role of SMAD4-deleted PDAC cells in modulating activation of stromal cell transformation is unknown. Our proposed aims here have aimed to bridge this knowledge gap. We propose a differential BCAA metabolism in cancer and stromal compartments of PDAC tumors and uncover a heavy reliance of PDAC cells on CAF-secreted branched chain keto acids (BCKAs) in stromal- rich tumors. We hypothesize that reactive stromal BCAA metabolism is altered from quiescent stroma and is the driver for regulating PDAC cell growth by secreting BCKAs. Second, SMAD4-deleted PDAC cells induces metabolic alterations in stromal cells. First, we will validate stromal BCKA synthesis metabolic pathway as a lethal target in patient-derived circulating tumor cell (CTC)-organoids and patient-derived tissue slices, using 13C-based isotope tracing and genetic tools and unravel that CAF-secreted extracellular matrix (ECM) is a source for BCKA. To understand the mechanism behind ECM degradation, we will regulate proteasomal proteolysis of ECM in CTC-organoids and patient-derived tissue slices. Second, to elucidate SMAD4-deletion in PDAC cells induced metabolic rewiring of CAFs we have proposed loss-of-function studies and use CTC- organoids and tissue slices to demonstrate increased stromal-reliance of SMAD4-deleted PDAC cells on stromal BCAT1 and elucidate temporal metabolic reprogramming in SMAD4-deleted CTC-organoids over time course of organoid formation. Third, we will test the efficacy of targeting BCKA anabolism in CAFs with conditional knockout mice and perform in vivo studies targeting stromal BCKA anabolic pathway using patient- derived low-passage cell lines and the syngeneic allograft model. Importantly, we will inhibit stromal BCAT1 in the proposed Aims using Gabapentin, Erg 240 (BCAT1 inhibitor from Ergon Pharma), ERG 245 (BCAT1/2 inhibitor from Ergon Pharma), & CRISPR targeting of BCAT1 in CAFs. Notably, our proposed aims will reveal a novel BCAA metabolism-centric regulatory role of reactive stroma in cancers and will uncover the underlying mode of action of this regulation. These findings can lead to novel therapeutics targeting communication between cancer cells and their microenvironment.

传统疗法在胰腺导管腺癌（PDAC）中的失败是由于我们有限的了解肿瘤微环境（TME）如何促进快速发展或复发 PDAC。在PDAC中，基质细胞被鉴定为癌症相关的成纤维细胞（CAF）肿瘤体积的90％。最近的研究揭示了分支链氨基酸的重要性（BCAA）在癌症中。但是，基质细胞在支持BCAA代谢中的作用仍然很差理解。了解癌症质体生态系统需要深入了解癌症的交集通过重新编程的BCAA代谢在基质中相关的转化。 smad4删除是 PDAC经常发生。但是，Smad4删除PDAC细胞在调节激活中的代谢作用基质细胞转化是未知的。我们提出的目标是弥合这一知识差距。我们提出了PDAC肿瘤的癌症和基质室中的BCAA代谢差异在基质 - 丰富的肿瘤。我们假设反应性基质BCAA代谢从静态基质改变了，并且IS 通过分泌BCKA来调节PDAC细胞生长的驱动因素。其次，Smad4删除的PDAC细胞诱导基质细胞的代谢改变。首先，我们将验证基质BCKA合成代谢途径为使用患者衍生的循环肿瘤细胞（CTC） - 孔子和患者衍生组织切片的致命靶标 13C基于CAF分泌的细胞外基质（ECM）的基于13C的同位素跟踪和遗传工具和漏洞是 BCKA的来源。要了解ECM降解背后的机制，我们将调节蛋白酶体 ECM在CTC-棒素和患者衍生的组织切片中的蛋白水解。其次，阐明SMAD4-DESTION 在PDAC细胞诱导CAF的代谢重新布线中，我们提出了功能丧失研究并使用CTC- 器官和组织切片以证明SMAD4删除PDAC细胞的基质依赖性增加随着时间的推移，SMAD4删除的CTC-甲型机构中的基质BCAT1和阐明时间代谢重编程器官形成的过程。第三，我们将测试靶向BCKA在CAF中的BCKA合成剂的功效有条件的敲除小鼠并进行体内研究，以患者为靶向基质BCKA合成途径派生的低通质细胞系和同种异体移植模型。重要的是，我们将在拟议的目的是使用Gabapentin，ERG 240（来自Ergon Pharma的BCAT1抑制剂），ERG 245（BCAT1/2） CAFS中BCAT1的Ergon Pharma的抑制剂和CRISPR靶向。值得注意的是，我们提出的目标将揭示新型BCAA代谢为中心的反应性基质在癌症中的作用，并将发现潜在的该法规的作用方式。这些发现可以导致新颖的治疗剂针对沟通在癌细胞和它们的微环境之间。