m⁶A modification regulates cell proliferation via reprogramming the balance between glycolysis and pentose phosphate...

N6-methyladenosine (m⁶A) stands as the predominant modification in eukaryotic mRNA and is involved in various biological functions. Aberrant m⁶A has been implicated in abnormal cellular phenotypes, including defects in stem cell differentiation and tumorigenesis. However, the precise effects of m⁶A on cell proliferation and the underlining mechanism of metabolic gene regulation remain incompletely understood. Here, we established a cellular environment with low-m⁶A levels and observed a severe impairment of cell proliferation. Mechanistic studies revealed that the depletion of m⁶A on TIGAR mRNA led to increased expression, subsequently inhibiting glycolysis while promoting the pentose phosphate pathway (PPP). A genome-wide CRISPR-Cas9 screen identified numerous genes involved in cell proliferation that are sensitive to m⁶A modification, with G6PD emerging as a key regulator. Integration of gene expression and survival data from cancer patients suggested that patients with elevated G6PD expression may exhibit enhanced responsiveness to tumor growth inhibition through m⁶A suppression. Our findings elucidate the critical role of m⁶A in cell proliferation, highlighting the therapeutic potential of targeting m⁶A-mediated metabolic pathways in cancer.