Epstein-Barr virus suppresses N 6 -Methyladenosine modification of TLR9 to promote immune evasion.

Epstein-Barr virus (EBV) is a human tumor virus associated with a variety of malignancies, including nasopharyngeal carcinoma, gastric cancers, and B-cell lymphomas. N6 -methyladenosine (m6 A) modifications modulate a wide range of cellular processes and participate in the regulation of virus-host cell interactions. Here, we discovered that EBV infection downregulates Toll-like receptor 9 (TLR9) m6 A modification levels and thus inhibits TLR9 expression. TLR9 has multiple m6 A modification sites. Knockdown of METTL3, an m6 A "writer", decreases TLR9 protein expression by inhibiting its mRNA stability. Mechanistically, Epstein-Barr nuclear antigen 1 (EBNA1) increases METTL3 protein degradation via K48-linked ubiquitin-proteasome pathway. Additionally, YTHDF1 was identified as an m6 A "reader" of TLR9, enhancing TLR9 expression by promoting mRNA translation in an m6 A -dependent manner, which suggests that EBV inhibits TLR9 translation by "hijacking" host m6 A modification mechanism. Using the METTL3 inhibitor STM2457 inhibits TLR9-induced B cell proliferation and Ig secretion, and opposes TLR9-induced immune responses to assist tumor cell immune escape. In clinical lymphoma samples, the expression of METTL3, YTHDF1 and TLR9 was highly correlated with immune cells infiltration. This study reveals a novel mechanism that EBV represses the important innate immunity molecule TLR9 through modulating the host m6 A modification system.