Targeting the TEA DNA binding Domain of TEAD with a Peptide Inhibitor TEAi Suppresses Oncogenic Transcription and Tumor Growth

The Hippo pathway is a critical regulator of organ size and tumorigenesis, with the TEAD-YAP/TAZ transcriptional complex serving as a key effector frequently hyperactivated in cancers. Targeting this complex has emerged as a promising anticancer strategy. Here, we develop TEAi, a 64-amino acid peptide derived from Drosophila Nerfin-1, which directly binds the TEA DNA-binding domain of TEAD. TEAi potently suppressed TEAD-YAP-driven transcriptional activity, as evidenced by luciferase reporter and qPCR assays targeting canonical downstream genes CTGF and CYR61. Mechanistically, TEAi inhibited the DNA-binding capacity of TEAD2 without direct DNA interaction, thereby abolishing promoter recruitment of the TEAD-YAP complex. unexpectedly, TEAi induced nuclear export and cytoplasmic accumulation of TEAD2, suggesting a non-canonical mechanism distinct from native Nerfin-1 function. Functionally, TEAi significantly suppressed cancer cell proliferation in vitro and tumor growth in vivo. Unlike inhibitors targeting the YAP-binding interface, TEAi directly blocks the DNA-binding activity of TEAD, offering a potential advantage by comprehensively inhibiting all TEAD-dependent oncogenic transcription. Our findings establish TEAi as a promising prototype for therapeutics targeting TEAD, with implications for treating cancers driven by Hippo pathway dysregulation.