A micro-metabolic rewiring assay for assessing hypoxia-associated cancer metabolic heterogeneity.
Publication Year:
2025
PubMed ID:
40093303
Funding Grants:
Public Summary:
Cancer cells can change the way they use energy, and these metabolic changes often make treatments less effective. Different tumors—and even different cells within the same tumor—can behave very differently, especially when they experience low oxygen levels, a condition known as hypoxia. Hypoxia is common in solid tumors and strongly influences how cancer cells grow, survive, and respond to drugs. However, scientists have lacked simple, scalable tools to study how individual cancer cells adjust their metabolism under hypoxic conditions.
In this study, we introduce a new laboratory method called the micro-metabolic rewiring (muMeRe) assay. This tool allows researchers to recreate low-oxygen conditions naturally produced by the cancer cells themselves, without needing complex equipment. This makes it possible to observe how different cancer cells generate hypoxia and shift their metabolism in response. We also created new ways to measure how flexible each cell type is in changing its metabolic behavior. Using this system, we tested a treatment designed to target cancer metabolism and showed that it can reduce drug resistance caused by hypoxia. Overall, this work provides a powerful new platform to help develop more effective, personalized cancer therapies.
Scientific Abstract:
Cancer metabolism plays an essential role in therapeutic resistance, where significant inter- and intra-tumoral heterogeneity exists. Hypoxia is a prominent driver of metabolic rewiring behaviors and drug responses. Recapitulating the hypoxic landscape in the tumor microenvironment thus offers unique insights into heterogeneity in metabolic rewiring and therapeutic responses, to inform better treatment strategies. There remains a lack of scalable tools that can readily interface with imaging platforms and resolve the heterogeneous behaviors in hypoxia-associated metabolic rewiring. Here we present a micro-metabolic rewiring (muMeRe) assay that provides the scalability and resolution needed to characterize the metabolic rewiring behaviors of different cancer cells in the context of hypoxic solid tumors. Our assay generates hypoxia through cellular metabolism without external gas controls, enabling the characterization of cell-specific intrinsic ability to drive hypoxia and undergo metabolic rewiring. We further developed quantitative metrics that measure the metabolic plasticity through phenotypes and gene expression. As a proof-of-concept, we evaluated the efficacy of a metabolism-targeting strategy in mitigating hypoxia- and metabolic rewiring-induced chemotherapeutic resistance. Our study and the scalable platform thus lay the foundation for designing more effective cancer treatments tailored toward specific metabolic rewiring behaviors.
