2 research outputs found
Pharmacological inhibition of CLK2 activates YAP by promoting alternative splicing of AMOTL2
Yes-associated protein (YAP), the downstream effector of the evolutionarily conserved Hippo pathway, promotes cellular proliferation and coordinates certain regenerative responses in mammals. Small molecule activators of YAP may, therefore, display therapeutic utility in treating disease states involving insufficient proliferative repair. From a high-throughput chemical screen of the comprehensive drug repurposing library ReFRAME, here we report the identification of SM04690, a clinical stage inhibitor of CLK2, as a potent activator of YAP-driven transcriptional activity in cells. CLK2 inhibition promotes alternative splicing of the Hippo pathway protein AMOTL2, producing an exon-skipped gene product that can no longer associate with membrane-bound proteins, resulting in decreased phosphorylation and membrane localization of YAP. This study reveals a novel mechanism by which pharmacological perturbation of alternative splicing inactivates the Hippo pathway and promotes YAP-dependent cellular growth
Covalent Targeting As a Common Mechanism for Inhibiting NLRP3 Inflammasome Assembly
The NLRP3 inflammasome is a cytosolic protein complex
important
for the regulation and secretion of inflammatory cytokines, including
IL-1β and IL-18. Aberrant overactivation of NLRP3 is implicated
in numerous inflammatory disorders. However, the activation and regulation
of NLRP3 inflammasome signaling remain poorly understood, limiting
our ability to develop pharmacologic approaches to target this important
inflammatory complex. Here, we developed and implemented a high-throughput
screen to identify compounds that inhibit the inflammasome assembly
and activity. From this screen, we identify and profile inflammasome
inhibition of 20 new covalent compounds across nine different chemical
scaffolds, as well as many known inflammasome covalent inhibitors.
Intriguingly, our results indicate that NLRP3 possesses numerous reactive
cysteines on multiple domains whose covalent targeting blocks the
activation of this inflammatory complex. Specifically, focusing on
compound VLX1570, which possesses multiple electrophilic moieties,
we demonstrate that this compound allows covalent, intermolecular
cross-linking of NLRP3 cysteines to inhibit inflammasome assembly.
Our results, along with the recent identification of numerous covalent
molecules that inhibit NLRP3 inflammasome activation, further support
the continued development of electrophilic compounds that target reactive
cysteine residues on NLRP3 to regulate its activation and activity