Effect of different Trp53 allelic states on the response to etoposide treatment in ER-Hoxb8 cells

Trp53 is an essential tumor suppressor gene that coordinates the response to cellular stress, including exposure to chemotherapeutic agents. We aimed to characterize the effect of various Trp53 allelic states (i.e. wild-type, monoallelic mutations and biallelic mutations) on the modulation of downstream pathways in the context of treatment with the chemotherapeutic agent etoposide. In order to obtain a cell model that represents key characteristics of the pre-malignant condition of clonal hematopoiesis, we utilized ER-Hoxb8 cells, which represent an immortalized, but otherwise genetically normal hematopoietic progenitor cell line. Generation of this cell model from a mouse strain carrying the inducible Trp53 missense mutation R245W in the endogenous locus and coupled to a GFP reporter, allows therefore to study the impact of the various allelic states of Trp53 on cellular consequences in a context that models clonal hematopoiesis. Er-Hoxb8 cells carrying the various Trp53 allelic configurations (wild-type: mono_GFPneg and bi_GFPneg; monoallelic mutated: mono_GFPpos; biallelic mutated: bi_GFPpos) were treated with DMSO or 10 uM etoposide for 3 hours, after which RNA was isolated and subjected to bulk RNA-seq.