Drugs modulating stochastic gene expression affect the erythroid differentiation process.

Anissa Guillemin,Ronan Duchesne,Fabien Crauste,Sandrine Gonin-Giraud,Olivier Gandrillon,Cristina Pina

doi:10.1371/journal.pone.0225166

Abstract

To better understand the mechanisms behind cells decision-making to differentiate, we assessed the influence of stochastic gene expression (SGE) modulation on the erythroid differentiation process. It has been suggested that stochastic gene expression has a role in cell fate decision-making which is revealed by single-cell analyses but studies dedicated to demonstrate the consistency of this link are still lacking. Recent observations showed that SGE significantly increased during differentiation and a few showed that an increase of the level of SGE is accompanied by an increase in the differentiation process. However, a consistent relation in both increasing and decreasing directions has never been shown in the same cellular system. Such demonstration would require to be able to experimentally manipulate simultaneously the level of SGE and cell differentiation in order to observe if cell behavior matches with the current theory. We identified three drugs that modulate SGE in primary erythroid progenitor cells. Both Artemisinin and Indomethacin decreased SGE and reduced the amount of differentiated cells. On the contrary, a third component called MB-3 simultaneously increased the level of SGE and the amount of differentiated cells. We then used a dynamical modelling approach which confirmed that differentiation rates were indeed affected by the drug treatment. Using single-cell analysis and modeling tools, we provide experimental evidence that, in a physiologically relevant cellular system, SGE is linked to differentiation.

Highlights

Cell-to-cell variability is intrinsic to all living forms, from prokaryotes [1, 2] to eukaryotes [3]
We recently described a surge in cell-to-cell variability that precedes an irreversible differentiation of normal primary chicken erythroid progenitors called T2EC [35], that is fully compatible with such a view [31]
In order to characterize the relation between stochastic gene expression (SGE) and differentiation, we first make sure to be able to change the amount of SGE in T2EC using three drug treatments: Artemisinin, Indomethacin and MB-3

Summary

Introduction

Cell-to-cell variability is intrinsic to all living forms, from prokaryotes [1, 2] to eukaryotes [3]. Such variability originates from many sources, but arguably stochastic gene expression (SGE) is an important driving force in the generation of cell-to-cell variability among genetically identical cells [4], additional regulation layers do exist [5]. Role of stochastic gene expression during differentiation process additional external funding received for this study. All these funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Methods

Results

Discussion

Conclusion