Inhibition of ZC3H13 Attenuates G2/M Arrest and Apoptosis by Alleviating NABP1 m6A Modification in AKI

Background: Acute kidney injury (AKI) refers to a clinical syndrome caused by various etiologies, leading to rapid decline in renal function in a short period of time. M6A modification, which is the most common internal modification of mRNA, have been reported to play a vital role in AKI. ZC3H13, a kind of m6A methyltransferases, plays vital roles in canceer. However, the biological roles of ZC3H13 in the pathogenesis of AKI is still unknown. Methods: Dot blot assay, and m6A ELISA were performed to detect the m6A modification in cisplatin-induced AKI mice and cisplatin-treated HK2 cells. Hematoxylin and eosin (H&E) and Periodic acid Schiff (PAS) were used to evaluate the histological and immunohistochemical change of the kidney. Flow cytometry and TUNEL staining were used to assess cell apoptosis rate in HK2 cells. Actinomycin was used to measure the stability of NABP1 mRNA. MeRIP-qPCR was used to detect the enrichment of NABP1 mRNA. RNA immunoprecipitation was performed to verify the binding of IGF2BP1 protein to NABP1 mRNA. Results: We observed that m6A modification and the expression of ZC3H13 were substantially elevated in cisplatin-induced AKI mice and cisplatin-treated HK2 cells. Gain- and loss-of-function experiments showed silence of ZC3H13 attenuated G2/M cell cycle arrest and apoptosis both in vivo and in vitro. Mechanistically, we demonstrated that ZC3H13 mediated cisplatin-induced cell cycle arrest, apoptosis, and renal injury by regulating NABP1. Moreover, we confirmed that m6A modification mediated by ZC3H13 stabilized NABP1 mRNA and was discriminated by IGF2BP1. Conclusion: In conclusion, ZC3H13 promoted the m6A modification of NABP1 and further enhanced its mRNA stability through IGF2BP1-dependent mechanism. The inhibition of ZC3H13 alleviated G2/M cell cycle arrest, apoptosis and kidney injury through regulating NABP1, showing that ZC3H13/NABP1 axis is a promising AKI treatment target. Funding: Government Support – Non-U.S.A simplified schematic diagram showing the effect of ZC3H13 in AKI.