The serine-threonine kinase TAO3 promotes cancer invasion and tumor growth by facilitating trafficking of endosomes containing the invadopodia scaffold TKS5α

Abstract

1Invadopodia are actin-based proteolytic membrane protrusions required for invasive behavior and 2 tumor growth. We used our high-content screening assay to identify kinases impacting 3 invadopodia formation. Among the top hits we selected TAO3, a STE20-like kinase of the GCK 4 subfamily, for further analysis. TAO3 was over-expressed in many human cancers, and regulated 5 invadopodia formation in melanoma, breast and bladder cancers. Furthermore, TAO3 catalytic 6 activity facilitated melanoma growth in 3-dimensional matrices and in vivo. We developed potent 7 catalytic inhibitors of TAO3 that inhibited invadopodia formation and function, and tumor cell 8 extravasation and growth. Using these inhibitors, we determined that TAO3 activity was required 9 for endosomal trafficking of TKS5α, an obligate invadopodia scaffold protein. A 10 phosphoproteomics screen for TAO3 substrates revealed the dynein subunit protein LIC2 as a 11 relevant substrate. Knockdown of LIC2 or expression of a phosphomimetic form promoted 12 invadopodia formation. Thus, TAO3 is a new therapeutic target with a distinct mechanism of 13 action. 14 15 SIGNIFICANCE 16Targeting tumor invasive behavior represents an understudied opportunity. We used an unbiased 17 screening approach to identify kinases required for invadopodia formation and function. We 18 validated TAO3, both genetically and with a novel inhibitor, and determined TAO3 function. Our