PO.MCB01.01 · 分子与细胞生物学
利用微流控质谱平台鉴定ER+乳腺癌对CDK4/6抑制剂耐药的独特代谢适应
Distinct metabolic adaptations to resistance to CDK4/6 inhibitors in ER+ breast cancer identified using microfluidic mass spectrometry platform
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：细胞周期蛋白依赖性激酶抑制剂（CDK4/6i）是近十年来雌激素受体阳性（ER+）乳腺癌治疗中最重要的、改变临床实践的进展之一。尽管取得了临床成功，CDK4/6i耐药仍是一项重大的临床挑战。重要的是，治疗反应与CDK4、CDK6或其他经典细胞周期蛋白的表达水平并不相关，提示耐药源于复杂的适应性机制。
方法：为应对这一挑战，我们从LCC2、LCC9和T47D乳腺癌细胞系中生成了对palbociclib和abemaciclib均耐受的药物耐受持留细胞群（DTPs）。对这些DTPs的转录组分析揭示了代谢酶表达的显著改变，提示代谢重编程在CDK4/6i耐受细胞的存活与维持中发挥核心作用。为进一步鉴定和定量代谢物及代谢通路的变化，我们使用一种动态采样平台（DSP）分析了这些DTP群体，该平台整合了样品导入机制、微加工处理装置以及电喷雾电离质谱（ESI-MS）分析。该装置采用先进的微加工技术制造，可实现实时多重处理与代谢谱分析的整合。全部非靶向代谢组学数据采用通路水平的基因集富集分析（GSEA）进行评估。
结果：在各耐药细胞状态中，多条代谢通路表现出与DTP样适应相关的协调性变化。palbociclib耐药的LCC2-DTPs表现出一种独特的代谢重塑轨迹，其特征为泛醌生物合成持续升高，而这一适应在abemaciclib耐药的LCC2-DTPs及其他DTP亚系中并不存在。这一药物特异性代谢特征提示palbociclib以独特方式作用于线粒体通路，可能影响治疗耐药。另一方面，涉及电子传递链和脂肪酸氧化的通路在abemaciclib的DTPs中上调，提示对每种CDK4/6i存在差异化的代谢调控。
结论：这些发现凸显了palbociclib和abemaciclib在内分泌耐药的ER+乳腺癌中造成的代谢后果存在差异，并强调了微流控ESI-MS工作流程在解析与治疗耐药和肿瘤进展相关的药物特异性适应状态方面的实用价值。
查看英文原文 English abstract
Background: Cyclin-dependent kinase inhibitors, CDK4/6i, have been one of the most significant practice-changing advances in the treatment of estrogen receptor positive (ER+) breast cancer in the recent decade. Despite their clinical success, CDK4/6i resistance remains a major clinical challenge. Importantly, therapeutic response does not correlate with expression levels of CDK4, CDK6, or other canonical cell cycle proteins, suggesting that resistance arises from complex adaptive mechanisms.
Methods: To address this challenge, we generated drug-tolerant persister cell populations (DTPs) from LCC2, LCC9 and T47D breast cancer cell lines to both palbociclib and abemaciclib. Transcriptomic profiling of these DTPs revealed significant alterations in the expression of metabolic enzymes, suggesting that metabolic reprogramming plays a central role in the survival and maintenance of CDK4/6i-tolerant cells. To further identify and quantify changes in metabolites and metabolic pathways, we analyzed these DTP populations using a dynamic sampling platform (DSP) integrating a sample introduction mechanism, a microfabricated processing device, and electrospray ionization mass spectrometry (ESI-MS) analysis. The device, manufactured via advanced microfabrication techniques, enables integration of real-time multiple processing and metabolic profiling. Full untargeted metabolomics data were evaluated using pathway level gene set enrichment analysis (GSEA).
Results: Across resistant cell states, multiple metabolic pathways exhibited coordinated shifts associated with DTP-like adaptation. Palbociclib resistant LCC2-DTPs exhibit a distinct metabolic remodeling trajectory characterized by consistently elevated ubiquinone biosynthesis, an adaptation absent in abemaciclib-resistant LCC2-DTPs and other DTP sublines. This drug-specific metabolic signature suggests that palbociclib engages mitochondrial pathways in a unique manner, potentially influencing therapeutic resistance. On the other hand, pathways involving electron transport chain and fatty acid oxidation were upregulated in abemaciclib DTPs, suggesting differential metabolic regulation in response to each CDK4/6i.
Conclusions: These findings highlight divergent metabolic consequences for palbociclib and abemaciclib in endocrine-resistant ER+ breast cancer and underscore the utility of microfluidic ESI-MS workflows for resolving drug-specific adaptive states relevant to therapeutic resistance and tumor progression.
利益披露 Disclosure
G. A. Slusher, None..
Y. Gu, None.
S. S. Badve,
Susan G. Komen for the Cure SAC220219 ).
CDMRP BCRP Level 1 BC241097 ).
1R01CA281932-01A1 ).
1R33CA297922-01 ).
A. G. Fedorov, None.
Y. Gökmen-Polar,
Susan G. Komen for the Cure SAC220219 ).
CDMRP BCRP Level 1 BC241097 ).
1R01CA281932-01A1 ).
1R33CA297922-01 ).