This MSCA-Global Fellowship, “Contribution of the Epstein-Barr Virus and the Tumour Microenvironment to Anti-Apoptotic Mechanisms in DLBCL” (CoMAnD), aimed to explore the synergistic role of the proto-oncogene MYC and the oncogenic Epstein-Barr virus (EBV) in the development and treatment of Diffuse large B cell Lymphoma (DLBCL). The post-doctoral fellow (PF) explored this by first generating novel models of DLBCL from primary human tonsillar germinal centre B cells through immortalisation with different proto-oncogenes (MYC, BCL-2, BCL-6) and/or EBV. These novel DLBCL models were then characterised, with a specific focus on 1) the impact of MYC and EBV on the expression of members of the BCL-2 family of proteins (members of the intrinsic apoptotic pathway) and 2) the sensitivity of the novel DLBCL models of BH3-mimetics, a family of small molecule inhibitors that act on pro-survival members of the BCL-2 family of proteins (WP1). Next, the impact of MYC and EBV on the evolution of the tumour microenvironment (TME) and pro-/anti-apoptotic signals was explored by transplanting the novel DLBCL models into humanised mouse models. These mice were treated with BH3-mimetics to target the intrinsic apoptotic pathway (WP2). Finally, the TME of the novel humanised mouse models of MYC and/or EBV+ DLBCL was compared to patient samples using ultrahigh multiplex immunohistochemistry to uncover the relationship between MYC and EBV, and the evolution of the TME and anti-/pro-apoptotic signalling in DLBCL (WP3).
This research project has significant societal impact as currently, EBV+ DLBCL is associated with worse survival outcomes than the EBV- counterpart. Despite this, no treatment stratification exists for patients with EBV+ DLBCL. Without tractable models of EBV+ DLBCL, developing novel therapeutic strategies is severely impeded. To address this barrier, this project used novel isogenic models of EBV+ and EBV- DLBCL, genetically engineered to express different cellular and viral oncogenes, recapitulating human disease. Beyond the scope of this project, which aimed to explore intrinsic apoptosis and the tumour microenvironment in EBV+ DLBCL, these models have the potential to be employed to explore a diverse range of mechanisms and pathways involved in tumour progression and treatment resistance in EBV+ DLBCL.
In summary, this project aimed to elucidate the relationship between EBV, the proto-oncogene MYC and their effects on the intrinsic apoptosis pathway and the tumour microenvironment. The PF explored the potential to treat EBV+ DLBCL by targeting the intrinsic apoptosis pathway using BH3-mimetic drugs and evaluated how EBV and MYC modulate the tumour microenvironment to promote therapy resistance. Towards these aims, this fellowship had 6 objectives, 3 scientific and 3 training and development:
O1: Explore how EBV infection and MYC synergise to transform B cells in vitro.
O2: Using humanised mouse models, determine how different viral and cellular oncogenic drivers impact the TME.
O3: Validate TME changes in primary tumours and explore how selected TME features regulate apoptosis.
O4: Project management.
O5: Training and knowledge transfer.
O6: Dissemination, exploitation and communication.
