结直肠癌 (CRC) 是一个多阶段过程，从良性腺瘤的形成开始，进展为浸润性癌，最后发生转移性扩散。肿瘤细胞必须调整其新陈代谢，以支持与疾病进展相关的能量和生物合成需求。因此，靶向癌细胞代谢是结直肠癌的一种有前途的治疗途径。然而，为了识别 CRC 阶段特有的代谢脆弱性的易处理节点，我们必须了解 CRC 发展过程中代谢如何变化。在这里，我们使用一个独特的模型系统 - 包括人类早期腺瘤到晚期腺癌。我们发现，腺瘤细胞在肿瘤进展的早期阶段会转变为升高的糖酵解，但仍维持氧化代谢。进展的腺癌细胞更多地依赖谷氨酰胺衍生的碳来为 TCA 循环提供燃料，而糖酵解和 TCA 循环活动在早期腺瘤细胞中仍然紧密耦合。腺癌细胞在燃料来源方面更加灵活，使它们能够在营养贫乏的环境中增殖。尽管具有这种可塑性，我们仍将天冬酰胺（ASN）合成确定为晚期腺癌细胞代谢脆弱性的一个节点。我们发现，天冬酰胺合成酶 (ASNS) 的缺失会阻碍其增殖，而早期腺瘤细胞对 ASN 缺失有很大抵抗力。从机制上讲，我们表明晚期腺癌细胞依赖 ASNS 来支持 mTORC1 信号传导以及最大糖酵解和氧化能力。早期腺瘤细胞对 ASNS 损失的抵抗可能是由于晚期细胞中不存在的反馈回路，使它们能够感知和调节 ASN 水平并通过自噬补充 ASN。总之，我们的研究定义了 CRC 发展过程中的代谢变化，并强调 ASN 合成是疾病后期的可靶向代谢脆弱性。版权所有 © 2024 作者。由 Elsevier GmbH 出版。保留所有权利。

Colorectal cancer (CRC) is a multi-stage process initiated through the formation of a benign adenoma, progressing to an invasive carcinoma and finally metastatic spread. Tumour cells must adapt their metabolism to support the energetic and biosynthetic demands associated with disease progression. As such, targeting cancer cell metabolism is a promising therapeutic avenue in CRC. However, to identify tractable nodes of metabolic vulnerability specific to CRC stage, we must understand how metabolism changes during CRC development. Here, we use a unique model system - comprising human early adenoma to late adenocarcinoma. We show that adenoma cells transition to elevated glycolysis at the early stages of tumour progression but maintain oxidative metabolism. Progressed adenocarcinoma cells rely more on glutamine-derived carbon to fuel the TCA cycle, whereas glycolysis and TCA cycle activity remain tightly coupled in early adenoma cells. Adenocarcinoma cells are more flexible with respect to fuel source, enabling them to proliferate in nutrient-poor environments. Despite this plasticity, we identify asparagine (ASN) synthesis as a node of metabolic vulnerability in late-stage adenocarcinoma cells. We show that loss of asparagine synthetase (ASNS) blocks their proliferation, whereas early adenoma cells are largely resistant to ASN deprivation. Mechanistically, we show that late-stage adenocarcinoma cells are dependent on ASNS to support mTORC1 signalling and maximal glycolytic and oxidative capacity. Resistance to ASNS loss in early adenoma cells is likely due to a feedback loop, absent in late-stage cells, allowing them to sense and regulate ASN levels and supplement ASN by autophagy. Together, our study defines metabolic changes during CRC development and highlights ASN synthesis as a targetable metabolic vulnerability in later stage disease.Copyright © 2024 The Authors. Published by Elsevier GmbH.. All rights reserved.