Data Independent Analysis proteomic characterisation of polycythaemia vera and Acute myeloid leukaemia

Lead Research Organisation: University of Manchester
Department Name: School of Medical Sciences

Abstract

Polycythaemia vis a myeloproliferative disease characterised by the accumulation of erythroid cells. This disease can progress to acute myeloid leukaemia. Despite advances in the understanding of myeloid neoplasia the aetiology of polycythaemia vera is poorly understood. The presence of the JAK2 mutation does not explain the heterogeneous clinical behaviour of the disease, and the molecular mechanisms and pathways underlying progression are not known.
Objectives:
We will use new technology platforms specifically designed for high resolution data-independent acquisition (DIA) and characterise the cellular and plasma constituents characteristic of healthy individuals, polycythaemia vera and acute myeloid leukaemia patients using the £18million MRC provided to build the Stoller Biomarker Discovery Centre and also the Waters research platforms.
Methods:
We will carry out analysis of the proteome of enriched primitive cells (nucleus and cytosol) and plasma from the peripheral blood using DIA (SWATH and Waters novel MSe and 2D-MS). In a single measurement, these techniques capture all of the components in a biological sample -a digital proteomic map. These maps allow for the iterative re-mining of the permanent digital record in silico. The top candidates identified as having significance in polycythaemia vera and acute myeloid leukaemia pathogenesis will be further validated in informatic analysis and then using drug discovery approaches. functional molecular biology studies.
Potential Outcomes:
Our proposed work programme will produce permanent digital maps allowing the investigation of disease processes using patient specific disease models to gain a better understanding of the processes underpinning the disease in the correct context. These models can be used in the development of new interventions to treat disease.
Fall back:
The application of DIA technologies to myeloproliferative neoplasms and acute leukaemias has never been carried out before and any knowledge gained from the project will be publishable. However, as a fall-back position, previous work on polycythaemia vera obtained by our group on the spliceosome pathway and polycythaemia vera will be exploited in cell biology assays. This work would suggest that there is an altered spliceosome complex and by consequence an altered proteome expression. We will carry out targeted/quantitative MS analysis to assess and quantify the changes in the levels of these proteins to see if they are molecular markers of the disease relevant to disease mechanism and molecular pathology.

Publications

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