RNA aided immunotherapy of Epstein-Barr virus associated cancers: A novel approach for reconstitution of tumor immunogenicity in 3D microfluidic chips.
Only 2% of the human genome is protein coding. The rest of the 98% of it is noncoding. In order to better understand the complexity which prevails in gene regulation, cell differentiation and proliferation, we must also focus on the noncoding part of our genome, both in health and and in disease.
The noncoding RNAs, previously considered part of the junk material, has now taken a central stage in gene regulation. The principal aim of this proposal is to identify how Epstein-Barr virus, the ubiquitous herpesvirus contributes to lymphoma development by altering microRNAs which affect immune checkpoints like PD-L1 and ICOSL. We have identified that EBNA2, a virally encoded nuclear protein can increase PD-L1 by downregulating the PD-L1 targeting miR-34a. As an economical viable and faster alternative to murine models, we will test 3D microfluidic chip based assays to assess immunogenicity of EBV infected lymphomas. In the same 3D microfluidic models, we will investigate if combined delivery of immune checkpoint inhibitor antibodies and anti PD-L1 miRNA such as miR-34a reconstitutes the reduced immunogenicity of lymphoma cells. The 3D microfluidic system will be used to cocultivate the tumor cells reconstituted with immune checkpoint inhibitors and miRNAs with activated T lymphocytes and as the readout of activated T cell responses, IFN-g and caspase 3 expression will be monitored. It is out hope that we will be able to establish a reliable and fast approach to investigate the potential of RNA aided cancer immunotherapy in the 3D microfluidic chip based system.