Repurposing Niclosamide for Targeting Pancreatic Cancer by Inhibiting Hh/Gli Non-Canonical Axis of Gsk3β

Abstract

Niclosamide (Nic), an FDA-approved anthelmintic drug, is reported to have anti-cancer efficacy and is being assessed in clinical trials for various solid tumors. Based on its ability to target multiple signaling pathways, in the present study, we evaluated the therapeutic efficacy of Nic on pancreatic cancer (PC) in vitro. We observed an anti-cancerous effect of this drug as shown by the G0/G1 phase cell cycle arrest, inhibition of PC cell viability, colony formation, and migration. Our results revealed the involvement of mitochondrial stress and mTORC1-dependent autophagy as the predominant players of Nic-induced PC cell death. Significant reduction of Nic-induced reactive oxygen species (ROS) and cell death in the presence of a selective autophagy inhibitor spautin-1 demonstrated autophagy as a major contributor to Nic-mediated cell death. Mechanistically, Nic inhibited the interaction between BCL2 and Beclin-1 that supported the crosstalk of autophagy and apoptosis. Further, Nic treatment resulted in Gsk3β inactivation by phosphorylating its Ser-9 residue leading to upregulation of Sufu and Gli3, thereby negatively impacting hedgehog signaling and cell survival. Nic induced autophagic cell death, and p-Gsk3b mediated Sufu/Gli3 cascade was further confirmed by Gsk3β activator, LY-294002, by rescuing inactivation of Hh signaling upon Nic treatment. These results suggested the involvement of a non-canonical mechanism of Hh signaling, where p-Gsk3β acts as a negative regulator of Hh/Gli1 cascade and a positive regulator of autophagy-mediated cell death. Overall, this study established the therapeutic efficacy of Nic for PC by targeting p-Gsk3β mediated non-canonical Hh signaling and promoting mTORC1-dependent autophagy and cell death.

Disturbance of the Warburg effect by dichloroacetate and niclosamide suppresses the growth of different sub-types of malignant pleural mesothelioma in vitro and in vivo

“…In addition, ROS is increased when DCA is combined with sorafenib ( Sun et al, 2021 ) or PX-478 ( Parczyk et al, 2021 ). Nic also increases ROS ( Zhou et al, 2017 ; Shangguan et al, 2020 ; Kaushal et al, 2021 ; Lohiya and Katti, 2021 ). In this study, DCA alone increased only hydrogen peroxide in 211H cells and superoxide in H226 cells.…”

Autophagy: A Key Player in Pancreatic Cancer Progression and a Potential Drug Target

“…Another interesting study involving PDAC assessed the effects of Spautin-1 with niclosamide, a compound known to inhibit cellular metabolism and induce apoptosis [ 355 ]. PDAC cells pre-incubated with Spautin-1 and subsequently treated with niclosamide resulted in markedly reduced apoptotic cell death [ 355 ].…”

“…Another interesting study involving PDAC assessed the effects of Spautin-1 with niclosamide, a compound known to inhibit cellular metabolism and induce apoptosis [ 355 ]. PDAC cells pre-incubated with Spautin-1 and subsequently treated with niclosamide resulted in markedly reduced apoptotic cell death [ 355 ]. Spautin-1 was found to reduce Beclin-1 levels which enabled abundant Bcl-2 to prevent BAX/BAK oligomerization and subsequently, reduced apoptosis [ 355 ].…”

“…PDAC cells pre-incubated with Spautin-1 and subsequently treated with niclosamide resulted in markedly reduced apoptotic cell death [ 355 ]. Spautin-1 was found to reduce Beclin-1 levels which enabled abundant Bcl-2 to prevent BAX/BAK oligomerization and subsequently, reduced apoptosis [ 355 ]. Notably, these anti-apoptotic effects from Spautin-1 need to be considered in apoptosis-induced PDAC therapy.…”

Drug Repurposing to Enhance Antitumor Response to PD-1/PD-L1 Immune Checkpoint Inhibitors

“…The anticancer mechanism of action of NCS is complex and multifactorial. Several molecular targets and pathways have been implicated, including degradation of β-catenin induced upon phosphorylation of glycogen synthase kinase-3 (GSK-3β) [ 80 , 81 , 82 ]. This small molecule can be combined with conventional cytotoxic agents such as camptothecin or temozolomide to treat glioblastoma [ 83 , 84 ], with paclitaxel or doxorubicin to treat triple-negative breast cancer [ 85 , 86 ], and with other drugs used to treat colon cancer, prostate cancer, osteosarcoma, etc.…”