Hematopoietic differentiation proceeds through a series of binary decisions where lymphoid and myeloid precursors are specified first, followed by more restricted bi- and monopotent progenitors. Each step involves extinction of the original gene expression program and establishment of a novel gene expression program. This is thought to be accomplished by both antagonistic and synergistic transcription factor interactions that establish cell type specific transcription factor combinations. Recent work from t he host laboratory demonstrated that enforced expression in B cells of either the C/EBPa or C/EBPb transcription factor reprograms these cells into macrophages by a series of paralell and sequential events that require endogenous PU.1. The proposed researc h project aims at dissecting the process of trans-differentiation in this system through analysing the changes that occur both at the level of gene expression and of transcription factor occupacy of target gene promoters. For these studies I will use B cel ls infected with an inducible form of C/EBP which will permit to observe the changes in gene expression over time, using Affymetrix-microarrays and quantitave PCR. To identify C/EBP and PU.1 target genes in B cells and macrophages I will use a "ChIP-on-chi p" approach. Results obtained should lead to a better understanding of the lymphoid and myeloid cell branching process and the molecular events accompanying reprogramming of one differentiated cell type into another.
Call for proposal
See other projects for this call