Establishing the potential of GSK3 manipulation for the treatment of human leukaemias

Background: Chronic lymphocytic leukaemia (CLL) is the most common leukaemia in the Western world. A number of effective treatments currently exist for symptomatic patients with CLL, but primary or acquired resistance to therapies means that the disease is incurable, and many experience considerable treatment-related toxicity. In contrast to CLL, the survival of older patients with the rarer haematological malignancy, acute myeloid leukaemia (AML) is poor and measureable in months. Therefore, there is a clear clinical need for novel interventions in both diseases.

Recent studies (including our own unpublished results) have indicated a role for mitochondrial activity as a significant regulator of cellular bioenergetics in CLL, and suggested a role for oxidative stress in the disease process. Pathological changes in Nrf2, Mcl-1 and Wnt/β-catenin signalling have been recognised in both CLL and AML, with all of these pathways sharing a common regulatory protein, namely GSK3. An emerging function of GSK3 is the control of protein stability, with more than 20 GSK3 substrates targeted for destruction following phosphorylation (https://www.ncbi.nlm.nih.gov/pubmed/29102676). We have a BBSRC funding application pending a decision, to comprehensively classify the GSK3 regulated proteome in metabolic tissues. However, the majority of the known targets control cell proliferation, thus it would seem sensible to expand that work to include neoplasia.
Therefore, we propose that GSK3 has specific protein targets that are dysregulated in both CLL and AML, and thus improving our understanding of the clinically relevant GSK3 targets in these diseases should provide opportunities for novel interventions, as monotherapy or in combination with existing treatments.

Aims: To classify the GSK3 target proteome in human CLL and AML cells, with a view to establishing the feasibility of developing novel interventions.

Methodology: We have cryopreserved CLL cells, plus isolated DNA, RNA and protein. We also have access to the most commonly used AML cell lines and primary human material for validation studies. Normal controls for comparative studies exist in the form of cryopreserved mobilised peripheral blood cell collections that can be fractionated to isolate haemopoietic stem cells and lymphoid populations. In collaboration with Dr Jeffrey Huang, Biomarker Service, we will use quantitative proteomics (10plex-ITRAQ) to compare the expression level of the cellular proteome in CLL and AML samples with appropriate controls. This will be compared with cells treated ± selective GSK3 inhibitors. We can also establish the mRNA levels of these targets to differentiate the targets regulated by protein stability from those regulated at the transcriptional level. From this we can determine biological pathways (eg mitochondrial, metabolic etc) that associate with clinical outcomes, with a view to develop novel strategies to combat these diseases.

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