Small-molecule inhibition of Lats kinases may promote Yap-dependent proliferation in postmitotic mammalian tissues

Nathaniel Kastan,Jeanne Chiaravalli,Theresa Alisch,Ksenia Gnedeva,Aleksandra A Petelski,Eldad Tzahor,Avraham Shakked,Neil Segil,Aaron Nagiel,A J Hudspeth,David J Huggins,Alla Aharanov

doi:10.1038/s41467-021-23395-3

Abstract

Hippo signaling is an evolutionarily conserved pathway that restricts growth and regeneration predominantly by suppressing the activity of the transcriptional coactivator Yap. Using a high-throughput phenotypic screen, we identified a potent and non-toxic activator of Yap. In vitro kinase assays show that the compound acts as an ATP-competitive inhibitor of Lats kinases—the core enzymes in Hippo signaling. The substance prevents Yap phosphorylation and induces proliferation of supporting cells in the murine inner ear, murine cardiomyocytes, and human Müller glia in retinal organoids. RNA sequencing indicates that the inhibitor reversibly activates the expression of transcriptional Yap targets: upon withdrawal, a subset of supporting-cell progeny exits the cell cycle and upregulates genes characteristic of sensory hair cells. Our results suggest that the pharmacological inhibition of Lats kinases may promote initial stages of the proliferative regeneration of hair cells, a process thought to be permanently suppressed in the adult mammalian inner ear.

Highlights

IntroductionRegeneration occurs either by activation and amplification of resident stem cells, as in the epithelia of the skin and intestine, or through cellular dedifferentiation and proliferation, as in the liver
Hippo signaling is an evolutionarily conserved pathway that restricts growth and regeneration predominantly by suppressing the activity of the transcriptional coactivator Yap
Every plate included a positive control, sub-confluent cells, and negative control, densely cultured cells, both exposed to the dimethyl sulfoxide (DMSO) vehicle

Summary

Introduction

Regeneration occurs either by activation and amplification of resident stem cells, as in the epithelia of the skin and intestine, or through cellular dedifferentiation and proliferation, as in the liver. In other instances, such as central nervous and cardiac-muscle tissues, cells exhibit little or no potential for regeneration after injury[1,2,3]. Genetic inactivation of Lats kinases is sufficient to drive cell-cycle reentry in the adult murine utricle and the chicken’s basilar papilla[22] These observations suggested that chemical activation of Yap signaling might engender supportingcell proliferation in adult tissue, a key missing step in the regeneration of the mammalian inner ear

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