Assays Used for Discovering Small Molecule Inhibitors of YAP Activity in Cancers

Abstract

YAP/TAZ are transcriptional coactivators that function as the key downstream effectors of Hippo signaling. They are commonly misregulated in most human cancers, which exhibit a higher level of expression and nuclear localization of YAP/TAZ, and display addiction to YAP-dependent transcription. In the nucleus, these coactivators associate with TEA domain transcription factors (TEAD1-4) to regulate the expression of genes that promote cell proliferation and inhibit cell death. Together, this results in an excessive growth of the cancerous tissue. Further, YAP/TAZ play a critical role in tumor metastasis and chemotherapy resistance by promoting cancer stem cell fate. Furthermore, they affect tumor immunity by promoting the expression of PD-L1. Thus, YAP plays an important role in multiple aspects of cancer biology and thus, provides a critical target for cancer therapy. Here we discuss various assays that are used for conducting high-throughput screens of small molecule libraries for hit identification, and subsequent hit validation for successful discovery of potent inhibitors of YAP-transcriptional activity. Furthermore, we describe the advantages and limitations of these assays.

Sestrin2 mediates FOXM1 expression to block the EMT process in non-small cell lung cancer through the AMPK/YAP pathway

“…However, there were few studies on the specific molecular regulatory mechanism of Sestrin2, which was worthy of more studies to be explore. YAP is a crucial molecule in Hippo signaling pathway which engaged in progression of cancers [33,34]. As expected, the expression and functions of YAP in NSCLC have been reported.…”

Development of a New HiBiT Biosensor Monitoring Stability of YAP/TAZ Proteins in Cells

“…In this study, we have developed novel ultra-stable HiBiT biosensors for monitoring the levels of YAP and TAZ in cells. These differ from the conventional split NanoLuc assay that depends on the interaction of target proteins for the complementation of the SmBiT and LgBiT fragments, which do not spontaneously interact with each other [50]. Many studies have exploited this conventional approach to monitor the activity of YAP/TAZ and their binding partners, mainly TEAD [51].…”

Proteomics appending a complementary dimension to precision oncotherapy

“…Upon phosphorylation, LATS1/2 and its splice protein MOB1A/B sequentially phosphorylated the downstream effector's YAP and TAZ. When the Hippo pathway was inhibited, unphosphorylated YAP/TAZ was released into the nucleus to form a complex with TEAD1–4 transcription factors that regulated the transcription of genes related to cell proliferation and survival [146 ] . Seo et al harnessed proteomic analysis to obtain a MAP4KS interacting protein, STRN4, and correlation analysis of its expression with YAP in endometrial cancer suggested STRN4 as a putative oncogenic factor in endometrial cancer [147 ] .…”