Epstein-Barr virus LMP1 manipulates the content and functions of extracellular vesicles to enhance metastatic potential of recipient cells

Abstract

Extracellular vesicles (EV) mediate intercellular communication events and alterations in normal vesicle content contribute to function and disease initiation or progression. The ability to package a variety of cargo and transmit molecular information between cells renders EVs important mediators of cell-to-cell crosstalk. Latent membrane protein 1 (LMP1) is a chief viral oncoprotein expressed in most Epstein-Barr virus (EBV)-associated cancers and is released from cells at high levels in EVs. LMP1 containing EVs have been demonstrated to promote cell growth, migration, differentiation, and regulate immune cell function. Despite these significant changes in recipient cells induced by LMP1 modified EVs, the mechanism how this viral oncogene modulates the recipient cells towards these phenotypes is not well understood. We hypothesize that LMP1 alters EV content and following uptake of the LMP1-modified EVs by the recipient cells results in the activation of cell signaling pathways and increased gene expression which modulates the biological properties of recipient cell towards a new phenotype. Our results show that LMP1 expression alters the EV protein and microRNA content packaged into EVs. The LMP1-modified EVs also enhance recipient cell adhesion, proliferation, migration, invasion concomitant with the activation of ERK, AKT, and NF-κB signaling pathways. The LMP1 containing EVs induced transcriptome reprogramming in the recipient cells by altering gene expression of different targets including cadherins, matrix metalloproteinases 9 (MMP9), MMP2 and integrin-α5 which contribute to extracellular matrix (ECM) remodeling. Altogether, our data demonstrate the mechanism in which LMP1-modified EVs reshape the tumor microenvironment by increasing gene expression of ECM interaction proteins.

Extracellular Vesicles as a Means of Viral Immune Evasion, CNS Invasion, and Glia-Induced Neurodegeneration

“…This decreased the ability of CD4+ T-cell EVs to inhibit HIV infection. In the context of EBV, the viral protein latent membrane protein-1 was shown to alter the protein and microRNA contents of EVs released from infected cells (Nkosi et al, 2020 ). Further, the altered EVs had an effect on several aspects of the target cells, including proliferation, adhesion, and migration.…”

Exosomes and organ-specific metastasis

“…47 LMP-1 is a major viral oncogene that is expressed in most Epstein-Barr (EB) virus-related cancers. 48,49 Nkosi et al 49 showed that LMP-1-modified extracellular vesicles can reshape the tumor microenvironment by changing the expression of different target genes including cadherin, MMP9, MMP2, and ITG a5. Wu et al 50 also found that LMP-1-modified extracellular vesicles promoted tumor proliferation and tumor PMN formation by activating CAFs.…”

Nanotechnology Frontiers in γ-Herpesviruses Treatments

“…Epstein–Barr nuclear antigens 1 and 2 (EBNA-1 and EBNA-2) are required to replicate the viral genome during the cell cycle progression, using the host DNA polymerase, and to gain a ‘persisting’ state, respectively [ 19 , 20 , 21 ]. In the last 60 years, several findings have showed that marmoset EBV-infected cells can proliferate and grow in vitro, expressing all of the latent proteins: six nuclear antigens (EBNA-1 to EBNA-6), latent membrane proteins (LMPs), and two non-polyadenylated RNAs (EBERs) [ 22 , 23 , 24 , 25 , 26 , 27 , 28 ]. One of them, LMP1, is an integral membrane protein that it mimics the CD40 receptor.…”

“…Immediate early (IE) proteins are necessary to trigger the viral replication expressing the early and late proteins. The former are required to replicate the viral genome in a defined structure, named concatemers, and released in a nucleocapsid, they gain the plasmatic membrane by a well-defined mechanism [ 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 ]. BFRF1 and BFLF2 encoded two proteins; they are essential to migrate and to translocate the incoming new virions in the cytoplasm by acquiring and losing the nuclear membrane shifts [ 31 , 32 , 33 , 34 ].…”