Theranostics 2018; 8(12):3400-3407. doi:10.7150/thno.24322

Research Paper

Molecular detection of inflammation in cell models using hyperpolarized 13C-pyruvate

Renuka Sriram, Julia Nguyen, Justin DeLos Santos, Linda Nguyen, Jinny Sun, Seth Vigneron, Mark Van Criekinge, John Kurhanewicz, John D. MacKenzie

Department of Radiology and Biomedical Imaging, University of California San Francisco, San Francisco, California, USA

Abstract

The detection and treatment monitoring of inflammatory states remain challenging in part due to the multifactorial mechanisms of immune activation and spectrum of clinical manifestations. Currently, diagnostic strategies tend to be subjective and limited quantitative tools exist to monitor optimal treatment strategies. Pro-inflammatory M1 polarized macrophages exhibit a distinct metabolic glycolytic phenotype compared to the continuum of M2 polarization states. In the present study, the distinct metabolic phenotypes of resting and activated macrophages were successfully characterized and quantified using hyperpolarized carbon-13 (13C) labeled pyruvate and its metabolic products, i.e. lactate, as a biomarker of resting, disease and treated states.

Methods: Mouse macrophage J774A.1 cells were used as a model system in an NMR compatible bioreactor to facilitate dynamic hyperpolarized 13C measurements. The glycolytic metabolism of the cells in the quiescent or resting state were compared with macrophages stimulated by lipopolysaccharide, a classical M1 activator using hyperpolarized 13C labeled pyruvate. Additionally, the activated macrophages were also treated with a non-steroidal anti-inflammatory drug to assess the changes in hyperpolarized lactate signal. The hyperpolarized lactate signals were then correlated using biochemical and molecular assays.

Results: We first validated our model system of inflammatory cells by the hallmarks of M1 polarization using steady state metabolic profiling with high resolution NMR in conjunction with nitric oxide Greiss assay, enzyme activity, and mRNA expression. Subsequently, we clearly showed that the cutting edge technology of hyperpolarized 13C NMR can be used to detect elevated lactate levels in M1 polarized macrophages in comparison to control and non-steroidal anti-inflammatory drug treated M2 states.

Conclusion: Hyperpolarized 13C lactate has the potential to serve as a biomarker to non-invasively detect and quantify pro-inflammatory state of immune regulatory cells and its response to therapy.

Keywords: hyperpolarized 13C nuclear magnetic resonance (MR), dynamic nuclear polarization (DNP), pyruvate, lactate, M1 polarization, macrophages, inflammation

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How to cite this article:
Sriram R, Nguyen J, Santos JD, Nguyen L, Sun J, Vigneron S, Van Criekinge M, Kurhanewicz J, MacKenzie JD. Molecular detection of inflammation in cell models using hyperpolarized 13C-pyruvate. Theranostics 2018; 8(12):3400-3407. doi:10.7150/thno.24322. Available from http://www.thno.org/v08p3400.htm