Optimized protocol for stable isotope tracing and steady-state metabolomics in mouse HER2+ breast cancer brain metastasis

Pravat Kumar Parida, Mauricio Marquez-Palencia, Akash K. Kaushik, Kangsan Kim, Vidhya Nair, Jessica Sudderth, Hieu Vu, Lauren Zacharias, Ralph DeBerardinis, Srinivas Malladi

Research output: Contribution to journalArticlepeer-review

Abstract

Analyzing the metabolic dependencies of tumor cells is vital for cancer diagnosis and treatment. Here, we describe a protocol for 13C-stable glucose and glutamine isotope tracing in mice HER2+ breast cancer brain metastatic lesions. We describe how to inject cancer cells intracardially to generate brain metastatic lesions in mice. We then detail how to perform 13C-stable isotope infusion in mice with established brain metastasis. Finally, we outline steps for sample collection, processing for metabolite extraction, and analyzing mass spectrometry data. For complete details on the use and execution of this protocol, please refer to Parida et al. (2022).

Original languageEnglish (US)
Article number101345
JournalSTAR Protocols
Volume3
Issue number2
DOIs
StatePublished - Jun 17 2022

Keywords

  • Cancer
  • Cell Biology
  • Cell culture
  • Mass Spectrometry
  • Metabolism
  • Metabolomics

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Medicine(all)

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