Programmable ZBTB7A RNA N6-methyladenosine site-specific demethylation suppresses colorectal cancer liver metastasis

Abstract

Colorectal cancer liver metastasis (CRLM) is the primary cause of CRC-related mortality, with inevitable chemoresistance to targeted therapies and immunotherapy. N 6 -methyladenosine (m 6 A), as a crucial epigenetic regulator of gene expression and cellular physiology, involved in the pathogenesis of CRLM. Precise manipulation of m 6 A modifications could offer a non-pharmacological precision treatment for many diseases. However, the precise editing of m 6 A modification in regulating CRLM progression remains elusive. Here we integrated multi-omics and identified zinc finger and BTB domain-containing 7A (ZBTB7A) as an m 6 A-modified transcription factor that promoted CRLM. Mechanistically, METTL3-mediated m 6 A modification of ZBTB7A facilitated recognition by the m 6 A reader YTHDF1/3 complex, enhancing its translation and expression. This m 6 A-dependent regulation promoted CRLM progression via activation of the ARHGAP26/Rho GTPase signaling axis. Notably, we applied a targeted RNA m 6 A erasure (TRME) system to achieve site-specific demethylation at a single site (m 6 A_site_411666) within ZBTB7A mRNA, without perturbing m 6 A abundance. Temporal demethylation at this site is sufficient to inhibit the CRC cell migration. This study unveils the critical role of the METTL3/ZBTB7A/ARHGAP26 axis in the process of m 6 A-mediated CRLM and positions m 6 A precise editing as a promising therapy in the preclinical treatment of CRLM.