Single-cell mass cytometry and transcriptome profiling reveal the impact of graphene on human immune cells

Marco Orecchioni,Kostas Kostarelos,Gianni Cesareni,Davide Bedognetti,Filomena Spada,Wouter Hendrickx,Cécilia Ménard-Moyon,Claudia Fuoco,Francesco Sgarrella,Artur Filipe Rodrigues,Lucia G Delogu,Leon Newman,Francesco M Marincola,Alberto Bianco

doi:10.1038/s41467-017-01015-3

Abstract

Understanding the biomolecular interactions between graphene and human immune cells is a prerequisite for its utilization as a diagnostic or therapeutic tool. To characterize the complex interactions between graphene and immune cells, we propose an integrative analytical pipeline encompassing the evaluation of molecular and cellular parameters. Herein, we use single-cell mass cytometry to dissect the effects of graphene oxide (GO) and GO functionalized with amino groups (GONH2) on 15 immune cell populations, interrogating 30 markers at the single-cell level. Next, the integration of single-cell mass cytometry with genome-wide transcriptome analysis shows that the amine groups reduce the perturbations caused by GO on cell metabolism and increase biocompatibility. Moreover, GONH2 polarizes T-cell and monocyte activation toward a T helper-1/M1 immune response. This study describes an innovative approach for the analysis of the effects of nanomaterials on distinct immune cells, laying the foundation for the incorporation of single-cell mass cytometry on the experimental pipeline.

Highlights

Understanding the biomolecular interactions between graphene and human immune cells is a prerequisite for its utilization as a diagnostic or therapeutic tool
Thin graphene oxide (GO) flakes and GO surfacefunctionalized with amino groups (GONH2)[35] via epoxide ring opening, using triethyleneglycol (TEG) diamine, were investigated
We have previously shown that the epoxide ring opening reaction is a versatile strategy to functionalize GO in a controlled manner

Summary

Introduction

Understanding the biomolecular interactions between graphene and human immune cells is a prerequisite for its utilization as a diagnostic or therapeutic tool. The functionalization of GO enhanced its biocompatibility toward the immune populations analyzed (Fig. 1b) with the exception of natural killer (NK) cells and memory CTLs (see Methods section for gating strategy), in which both GO and GONH2 induced minimal cytotoxicity.

Results

Conclusion