MET TL3 drives telomere targ eting of TERRA lncRNA through m 6 A-dependent R-loop formation: a therapeutic target for ALT-positive neuroblastoma

Abstract

Telomerase-negative tumors maintain telomere length by alternative lengthening of telomeres (ALT), but the underlying mechanism behind ALT remains poorly understood. A proportion of aggressive neuroblastoma (NB), particularly relapsed tumors, are positiv e f or AL T (AL T+), suggesting that a better dissection of the ALT mechanism could lead to no v el therapeutic opportunities. TERRA, a long non-coding RNA (lncRNA) derived from telomere ends, localizes to telomeres in a R-loop-dependent manner and pla y s a crucial role in telomere maintenance. Here we present evidence that RNA modification at the N 6 position of internal adenosine (m 6 A) in TERRA b y the meth yltransferase METTL3 is essential f or telomere maintenance in ALT+ cells, and the loss of TERRA m 6 A / METTL3 results in telomere damage. We observed that m 6 A modification is abundant in R-loop enriched TERRA, and the m 6 A-mediated recruitment of hnRNPA2B1 to TERRA is critical for R-loop formation. Our findings suggest that m 6 A drives telomere targeting of TERRA via R-loops, and this m 6 A-mediated R-loop formation could be a widespread mechanism emplo y ed b y other chromatin-interacting lncRNAs. Furthermore, treatment of ALT+ NB cells with a METTL3 inhibitor resulted in compromised telomere targeting of TERRA and accumulation of DNA damage at telomeres, indicating that METTL3 inhibition may represent a therapeutic approach for ALT+ NB.