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The role of MSCs derived inflammation in the pathogenesis of Myeloproliferative Neoplasms (MPN)

The aim of this study is to further investigate the phenotypic and functional differences between MPN-MSCs, healthy donor MSCs as well as 'inflamed' MSCs following multiple TLR agonist stimulation.

Applications are now closed for the 2019 postgraduate training programme.

  • Primary Supervisor: Dr Dominique Bonnet, the Crick
  • Secondary Supervisor: Dr Shahram Kordasti, Systems Cancer Immunology, King’s College London

Funding note: Non-EU candidates are not eligible to apply

Background

Chronic inflammation is known to increase the risk of genomic instability and malignant transformation. Activation of signal transducer and activator of transcription (STAT) family members, particularly STAT3, is considered one of the most important pathways for inflammation-mediated tumourigenesis. Cancer-mediated inflammation not only has anti-apoptotic effects, it also increases cell proliferation and subsequently leads to genomic destabilization and somatic mutations. Myeloproliferative neoplasms (MPNs) are acquired stem cell neoplasms that display autonomous proliferative advantages associated with JAK/STAT pathway deregulation. All MPNs are associated with myeloid-derived inflammation and somatic mutations such as JAK2V617F and CALR can be detected in the majority of patients. MPNs could also evolve into acute leukaemia. Additional somatic mutations of additional genes such as RUNX1, TP53, EZH2, and LNK are considered as independent risk factor for leukemic transformation [1]. However, the source of inflammation and its effect on both adaptive immune response as well as growth advantage of mutated clones are less clear. Interestingly, Mesenchymal Stromal Cells (MSCs) originated from JAK2V617F mutated patients overexpress SPP1 and NF-kB and could support CFU-GM colonies [2].

Project detail

The role of MSCs derived inflammation in the pathogenesis of Myeloproliferative Neoplasms (MPN) and leukaemic transformation.

The aim of this study is to further investigate the phenotypic and functional differences between MPN-MSCs, healthy donor MSCs as well as “inflamed” MSCs following multiple TLR agonist stimulation. We will also compare the difference between these MSCs in supporting MPN propagating cells. State of the art techniques such as mass cytometry (known as cytometry by time-of-flight (CyTOF)), deep sequencing for somatic mutations, single-cell RNAsequencing (scRNAseq) as well as ex vivo and in vivo functional assays will be used for this project. This will help us to identify novel biotherapeutic targets which is an unmet need in MPN.

The specific aims and methodology of this studentship are:

  1. Cross talk between MPN-MSCs, adaptive immune cells and MPN-Propagating cells: CD34+ cells from JAK2V617F mutated patients will be co-cultured for up to 7 days in the presence either healthy donor (HD)MSCs (with and without stimulation by combined TLR agonists) or MPN-MSCs. Following culture, scRNAseq will be performed on both MPN cells and MSCs fractions. The PhD student will be working closely with our bioinformatics team to customise and implement new tools for data analysis and develop unbiased and automated analysis pipelines.
  2. Animal model: In parallel, we will use our humanised 3D scaffold model [3] to dissect the cross-talk between MPN-propagating cells and MSCs. Either HD-MSC or autologous MPN-MSC will be used. 10-14 weeks after implantation of the scaffold, we will retrieve the MPN cells and performed NGS and/or target sequencing to analysis the clonal composition of the MPN and compared this to the original patients. We will also evaluate whether MPN-MSC were or not able to migrate to the mouse BM. We will extract the MSC fractions and performed scRNAseq. This analysis will be compared to our ex vivo data analysis. 
  3. We will validate some of the potential pathways identified using MPN– bone marrow samples and the functional effects of inhibition/activation

Potential research placements:

  1. Familiarisation of computational tools and skills. Translational Systems Biology Group, Guy’s Campus, King’s College London, supervised by Dr Saeed Shoaie.
  2. Learning how to culture, stimulate and analyse the phenotype of MSCs. Regenerative Medicine Lab, Denmark Hill Campus, King’s College London, supervised by Prof Francesco Dazzi.    
  3. Working with 3D materials and analysing MSC behaviours. Craniofacial Development and Stem Cell Biology Group, Guy’s Campus, King’s College London, supervised by Dr Eileen Gentleman.

References

  1. Lasho TL, Mudireddy M, Finke CM, et al. Targeted next-generation sequencing in  blast phase myeloproliferative neoplasms. Blood Adv, 2018;2:370-80.
  2. Ramos TL et al. Mesenchymal stromal cells (MSC) from JAK2+ myeloproliferative neoplasms differ from normal MSC and contribute to the maintenance of neoplastic hematopoiesis. PLoS One, 2017;12:e0182470.
  3. Abarrategi A et al. Versatile humanized niche model enables study of normal and malignant human hematopoiesis. J Clin Invest. 2017;127(2):543-548.