The role of PHF6 during haematopoietic differentiation

PHD finger protein 6 (PHF6) is a protein, consisting of two atypical plant homeodomains (aPHD1/2) with nuclear and nucleolar localization sequences. It is mutated in a variety of different leukaemia, including T-cell acute lymphoblastic leukaemia (T-ALL), acute myeloid leukaemia (AML), and mixed phenotype acute leukaemia (MPAL). The mutations identified in leukaemia are predominantly nonsense and frame shifts, resulting in the absence of protein expression.

A number of interactions of PHF6 with other proteins have been described, but mostly in non-haematological cell types. Recently we reported the interaction between PHF6 and the LMO2 complex. The LMO2 complex is a haematopoietic transcriptional regulator involving a number of transcription factors and mediators. It is known to be required from the onset of haematopoietic development, but is downregulated in terminally differentiating cells. In this project we will study the differentiation of in the presence and absence of LMO2 and/or PHF6. We will perform these experiments using embryonic stem cell differentiation to study this during the onset of haematopoiesis and using CD34+ haematopoietic stem/progenitor cells to study this during haematopoietic differentiation and lineage commitment. Techniques that will be utilised include, tissue culture, Western blot, mRNA expression analysis, quantitative PCR, CRISPR-Cas9 and shRNA knockdown, chromatin immunoprecipitation, immunohistochemistry, genome wide sequencing.