NSD2 Promotes Tumor Angiogenesis Through Methylating and Activating STAT3 Signaling

Background：Tumor angiogenesis plays important roles in tumorigenesis and development, the regulation mechanism of angiogenesis is still not been fully elucidated. Nuclear receptor binding SET domain protein 2 (NSD2), a histone methyltransferase which catalyzes the di-methylation of histone H3 at lysine 36, has been proved a critical molecule in proliferation, metastasis and tumorigenesis. But its role in tumor angiogenesis remains unknown.Methods： Cell Counting Kit 8 (CCK8), scratch assays, transwell-migration assays, tube-formation and mice xenograft model assays were used to confirm the role of NSD2 in the bioprocess of angiogenesis. Bioinformatics analysis, western blot and immunofluorescence staining were used to verify the function of NSD2 in regulating STAT3 signaling pathway. Immunofluorescence co-localization, immunoprecipitation, mass spectrometry and site-directed mutagenesis were performed to determine the NSD2-dependent methylation site of STAT3.Results: Here we demonstrated that NSD2 promoted tumor angiogenesis in vitro and in vivo. Furthermore, we confirmed that the angiogenic function of NSD2 was mediated by STAT3. Momentously, we found that NSD2 promoted the methylation of STAT3 and that the inhibition of STAT3 methylation resulted in the attenuation of STAT3 signaling pathway. In addition, mass spectrometry and site-directed mutagenesis assays revealed that NSD2 methylated STAT3 at lysine 163 (K163), and K163 was of significance in the activation of STAT3 signaling pathway.Conclusion: We conclude that methylation of STAT3 catalyzed by NSD2 promotes the activation of STAT3 pathway and enhances the ability of tumor angiogenesis. Our findings investigate a NSD2 dependent methylation-phosphorylation regulation pattern of STAT3 and reveal that NSD2/STAT3/VEGFA axis might be a potential target for tumor therapy.

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