Covalent disruptor of YAP-TEAD association suppresses defective Hippo signaling

Our plain-English summary of Covalent disruptor of YAP-TEAD association suppresses defective Hippo signaling, by M. Fan, W. Lu, J. Che, et al (eLife, 2022).

The study

• Authors: M. Fan, W. Lu, J. Che, N.P. Kwiatkowski, Y. Gao, H.S. Seo, S.B. Ficarro, P.C. Gokhale, Y. Liu, E.A. Geffken, J. Lakhani, K. Song, M. Kuljanin, W. Ji, J. Jiang, Z. He, J. Tse, A.S. Boghossian, M.G. Rees, M.M. Ronan, J.A. Roth, J.D. Mancias, J.A. Marto, S. Dhe-Paganon, T. Zhang, N.S. Gray
• Journal: eLife
• Published: 2022;11
• Cited by: 58 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

The transcription factor TEAD, together with its coactivator YAP/TAZ, is a key transcriptional modulator of the Hippo pathway. Activation of TEAD transcription by YAP has been implicated in a number of malignancies, and this complex represents a promising target for drug discovery. However, both YAP and its extensive binding interfaces to TEAD have been difficult to address using small molecules, mainly due to a lack of druggable pockets. TEAD is post-translationally modified by palmitoylation that targets a conserved cysteine at a central pocket, which provides an opportunity to develop cysteine-directed covalent small molecules for TEAD inhibition. Here, we employed covalent fragment screening approach followed by structure-based design to develop an irreversible TEAD inhibitor MYF-03-69. Using a range of in vitro and cell-based assays we demonstrated that through a covalent binding with TEAD palmitate pocket, MYF-03-69 disrupts YAP-TEAD association, suppresses TEAD transcriptional activity and inhibits cell growth of Hippo signaling defective malignant pleural mesothelioma (MPM). Further, a cell viability screening with a panel of 903 cancer cell lines indicated a high correlation between TEAD-YAP dependency and the sensitivity to MYF-03-69. Transcription profiling identified the upregulation of proapoptotic BMF gene in cancer cells that are sensitive to TEAD inhibition. Further optimization of MYF-03-69 led to an in vivo compatible compound MYF-03-176, which shows strong antitumor efficacy in MPM mouse xenograft model via oral administration. Taken together, we disclosed a story of the development of covalent TEAD inhibitors and its high therapeutic potential for clinic treatment for the cancers that are driven by TEAD-YAP alteration.

© 2022, Fan, Lu et al.

In plain English

This was a laboratory drug-discovery study, not a study of people. The authors screened a biased library of cysteine-reactive covalent fragments by mass spectrometry against recombinant TEAD2 and identified MYF-01-37, which labelled cysteine 380. Structure-based optimisation produced MYF-03-69, which labelled TEAD2 completely at a 10-fold molar excess and, in a 1.68 Å co-crystal with TEAD1, formed a covalent bond with Cys359 in the palmitate pocket. MYF-03-69 inhibited palmitoylation of all four TEAD paralogs at submicromolar IC50 values, while the non-covalent analogue MYF-03-69-NC was inactive. In NCI-H226 mesothelioma cells it reduced co-immunoprecipitation of YAP with TEAD, inhibited a TEAD reporter (IC50 56 nM) and, at 2 µM, altered 339 genes. It slowed growth of NCI-H226 and MSTO-211H cells but not MeT-5A or NCI-H2452 cells. Across 903 barcoded cancer cell lines, 33 lines had AUC of 0.8 or less, mostly YAP- or TEAD-dependent. The orally dosed analogue MYF-03-176 produced average tumour regressions of 54 percent (30 mg/kg) and 68 percent (75 mg/kg) in NCI-H226 xenograft mice.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory drug-discovery study: covalent fragment screening, protein crystallography, biochemical and cell-based assays, a multi-cell-line viability screen, and a mouse xenograft experiment
• Who was studied: Recombinant TEAD1-4 YAP-binding domain proteins; human cell lines including NCI-H226, MSTO-211H, NCI-H2452 mesothelioma cells, MeT-5A mesothelium cells, HEK293T and others; 903 barcoded cancer cell lines in the PRISM screen; female 7-week-old NSG mice bearing NCI-H226 xenografts (8-9 mice per group), work conducted at Dana-Farber Cancer Institute
• Exposure: The covalent TEAD inhibitors MYF-01-37, MYF-03-69 and its orally bioavailable analogue MYF-03-176, with the non-covalent control MYF-03-69-NC and comparator TEAD palmitate-pocket binders K-975, compound 2 and VT103; mice received MYF-03-176 by oral gavage twice daily at 30 or 75 mg/kg for 28 days
• Main results:
  • MYF-03-69 at a 10-fold molar excess fully labelled recombinant TEAD2, with the modified residue identified as C380
  • A co-crystal structure of TEAD1 YBD with MYF-03-69 at 1.68 Å showed a covalent bond to Cys359
  • MYF-03-69 inhibited palmitoylation of all four recombinant TEAD paralogs at submicromolar IC50 values, whereas the non-covalent analogue MYF-03-69-NC did not
  • In NCI-H226 mesothelioma cells, 24 hr of MYF-03-69 reduced endogenous YAP pulled down with a pan-TEAD antibody, with little change in TEAD protein level
  • In NCI-H226 reporter cells, MYF-03-69 reduced YAP-TEAD transcriptional activity with an IC50 of 56 nM over 72 hr
  • In cysteine proteome profiling of NCI-H226 cells, 7 of 12,498 mapped cysteines showed more than 50 percent conjugation at 25 µM MYF-03-69; the TEAD palmitate pocket cysteines were not detected
  • RNA sequencing of NCI-H226 cells found 339 genes differentially expressed at 2 µM, while at 0.1 µM only CTGF changed significantly
  • In 5-day growth assays MYF-03-69 inhibited NCI-H226 and MSTO-211H cells but not MeT-5A mesothelium cells or NCI-H2452 mesothelioma cells with intact Hippo signalling
  • In the PRISM screen of 903 cancer cell lines, 33 lines met a sensitivity threshold of AUC of 0.8 or less and most were YAP- or TEAD-dependent by CERES scores
  • BMF mRNA rose in the three sensitive lines but not in the three insensitive lines, although it also rose in PC9 and HEK293T cells, which were resistant
  • In the NCI-H226 xenograft mice, oral MYF-03-176 twice daily for 28 days gave average tumour regression of 54 percent at 30 mg/kg and 68 percent at 75 mg/kg, a difference the authors reported as not significant
  • At 75 mg/kg mice lost on average 5 percent body weight and three of eight animals lost 12-14 percent, which recovered after dosing stopped
• The authors' conclusion: The authors concluded that MYF-03-69 is a potent covalent pan-TEAD inhibitor that disrupts YAP-TEAD association and alters transcription in a way dependent on covalent binding, and that they nominate it as a lead for drug discovery in mesothelioma and other YAP- or TEAD-dependent cancers
• Limitations the authors state:
  • The TEAD palmitate pocket cysteines were not detected by the DBIA probe used for proteome-wide selectivity profiling, possibly because of low TEAD abundance or because the probe can only be used on cell lysate
  • The authors stated that the mechanism by which palmitate-pocket binders inhibit TEAD transcription remains unknown
  • Upregulation of BMF did not track perfectly with sensitivity, since it also increased in PC9 and HEK293T cells that resisted TEAD inhibition; the authors offered signalling heterogeneity as a possible explanation

Topics

Medical terms in this study, explained in our Lexicon: Discovery, Pleural Mesothelioma.

Related studies

• Direct and selective pharmacological disruption of the YAP-TEAD interface by IAG933 inhibits Hippo-dependent and RAS-MAPK-altered cancers — E.A. Chapeau, L. Sansregret, et al. (2024)
• LIM-domain protein AJUBA suppresses malignant mesothelioma cell proliferation via Hippo signaling cascade — I. Tanaka, H. Osada, et al. (2015)
• Discovery of a Potent and Selective TEAD Degrader with Durable Degradation Activity — L. Cao, J. Yang, et al. (2025)
• First-in-human phase Ia study of the covalent TEAD palmitoylation inhibitor BPI-460372 in patients with advanced solid tumors: pharmacokinetics, exposure-limited dosing, and early antitumor activity — H. Liang, Y. Xu, et al. (2026)
• TAZ activation by Hippo pathway dysregulation induces cytokine gene expression and promotes mesothelial cell transformation — A. Matsushita, T. Sato, et al. (2019)
• Comparative Assessment and High-Throughput Drug-Combination Profiling of TEAD-Palmitoylation Inhibitors in Hippo Pathway Deficient Mesothelioma — L. Evsen, P.J. Morris, et al. (2023)

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