Inductively-Coupled Plasma Mass Spectrometry-Novel Insights From an Old Technology Into Stressed Red Blood Cell Physiology

Daniel Stephenson; Travis Nemkov; Syed M Qadri; William P Sheffield; Angelo D'Alessandro

doi:10.3389/fphys.2022.828087

Inductively-Coupled Plasma Mass Spectrometry-Novel Insights From an Old Technology Into Stressed Red Blood Cell Physiology

Front Physiol. 2022 Feb 7:13:828087. doi: 10.3389/fphys.2022.828087. eCollection 2022.

Authors

Daniel Stephenson¹, Travis Nemkov¹, Syed M Qadri², William P Sheffield^{3

4}, Angelo D'Alessandro¹

Affiliations

¹ Department of Biochemistry and Molecular Genetics, University of Colorado Denver-Anschutz Medical Campus, Aurora, CO, United States.
² Faculty of Health Sciences, Ontario Tech University, Oshawa, ON, Canada.
³ Centre for Innovation, Canadian Blood Services, Hamilton, ON, Canada.
⁴ Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON, Canada.

Abstract

Background: Ion and metal homeostasis are critical to red blood cell physiology and Inductively Coupled Plasma (ICP) is a decades old approach to pursue elemental analysis. Recent evolution of ICP has resulted in its coupling to mass spectrometry (MS) instead of atomic absorption/emission.

Methods: Here we performed Inductively-coupled plasma mass spectrometry (ICP-MS) measurements of intra- and extra-cellular Na, K, Ca, Mg, Fe, and Cu in red blood cells undergoing ionic, heat, or starvation stress. Results were correlated with Ca measurements from other common platforms (e.g., fluorescence-based approaches) and extensive measurements of red blood cell metabolism.

Results: All stresses induced significant intra- and extracellular alterations of all measured elements. In particular, ionomycin treatment or hypertonic stress significantly impacted intracellular sodium and extracellular potassium and magnesium levels. Iron efflux was observed as a function of temperatures, with ionic and heat stress at 40°C causing the maximum decrease in intracellular iron pools and increases in the supernatants. Strong positive correlation was observed between calcium measurements via ICP-MS and fluorescence-based approaches. Correlation analyses with metabolomics data showed a strong positive association between extracellular calcium and intracellular sodium or magnesium levels and intracellular glycolysis. Extracellular potassium or iron were positively correlated with free fatty acids (especially mono-, poly-, and highly-unsaturated or odd-chain fatty acid products of lipid peroxidation). Intracellular iron was instead positively correlated with saturated fatty acids (palmitate, stearate) and negatively with methionine metabolism (methionine, S-adenosylmethionine), phosphatidylserine exposure and glycolysis.

Conclusion: In the era of omics approaches, ICP-MS affords a comprehensive characterization of intracellular elements that provide direct insights on red blood cell physiology and represent meaningful covariates for data generated via other omics platforms such as metabolomics.

Keywords: RBC death; calcium; inductively coupled plasma; iron; metabolomics; potassium; sodium.

Abstract

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