The m6A demethylase ALKBH5 promotes tumor progression by inhibiting RIG-I expression and interferon alpha production through the IKKε/TBK1/IRF3 pathway in head and neck squamous cell carcinoma

• Main Authors: Chang, H. (Author), He, Y. (Author), Jin, S. (Author), Li, M. (Author), Ma, H. (Author), Wang, R. (Author), Zhang, J. (Author), Zhang, Z. (Author)
• Format: Article
• Language: English
• Published: BioMed Central Ltd 2022
• Subjects: AlkB Homolog 5, RNA Demethylase; ALKBH5; ALKBH5 protein, human; alkylated DNA repair protein alkB homolog 5; alpha interferon; animal; Animals; C3H mouse; DEAD Box Protein 58; genetics; Head and Neck Neoplasms; Head and neck squamous cell carcinoma; head and neck tumor; human; Humans; I kappa B kinase; I-kappa B Kinase; Interferon alpha; interferon regulatory factor 3; Interferon Regulatory Factor-3; Interferon-alpha; IRF3 protein, human; Irf3 protein, mouse; messenger RNA; metabolism; Mice; Mice, Inbred C3H; mouse; N6-methyladenosine; Protein Serine-Threonine Kinases; retinoic acid inducible protein I; RIG-I; RNA, Messenger; Squamous Cell Carcinoma of Head and Neck; TBK1 protein, human; Tbk1 protein, mouse; tumor microenvironment; Tumor Microenvironment
• Online Access: View Fulltext in Publisher

 Background: N6-methyladenosine (m6A) RNA modification plays a critical role in various physiological and pathological conditions. However, the role of m6A modification in head and neck squamous cell carcinoma (HNSCC) remains elusive. Methods: In this study, the expression of m6A demethylases was detected by HNSCC tissue microarray. m6A-RNA immunoprecipitation (MeRIP) sequencing and RNA sequencing were used to identify downstream targets of ALKBH5. Comprehensive identification of RNA-binding proteins by mass spectrometry (ChIRP-MS) was used to explore the m6A “readers”. Tumor-infiltrating lymphocytes were analyzed in SCC7-bearing xenografts in C3H mice. Results: Here, we demonstrate the downregulation of m6A status and upregulation of two demethylases in HNSCC. Silencing the m6A demethylase alkB homolog 5, RNA demethylase (ALKBH5) suppresses tumor progression in vitro and in vivo. m6A-RNA immunoprecipitation sequencing reveals that ALKBH5 downregulates the m6A modification of DDX58 mRNA. Moreover, RIG-I, encoded by the DDX58 mRNA, reverses the protumorigenic characteristics of ALKBH5. ChIRP-MS demonstrates that HNRNPC binds to the m6A sites of DDX58 mRNA to promote its maturation. ALKBH5 overexpression inhibits RIG-I-mediated IFNα secretion through the IKKε/TBK1/IRF3 pathway. The number of tumor-infiltrating lymphocytes in C3H immunocompetent mice is reduced by ALKBH5 overexpression and restored by IFNα administration. Upregulation of AKLBH5 negatively correlates with RIG-I and IFNα expression in HNSCC patients. Conclusions: These findings unveil a novel mechanism of immune microenvironment regulation mediated by m6A modification through the ALKBH5/RIG-I/IFNα axis, providing a rationale for therapeutically targeting epitranscriptomic modulators in HNSCC. © 2022, The Author(s).