AMPK activation by ASP4132 inhibits non-small cell lung cancer cell growth

Ying-Chen Xia,Jian-Hua Zha,Ben-Tong Yu,Yan Zhang,Jie Zhen,Hui Yin,Yong-Hua Sang,Guo-Qiu Xu

doi:10.1038/s41419-021-03655-2

Abstract

Activation of adenosine monophosphate-activated protein kinase (AMPK) is able to produce significant anti-non-small cell lung cancer (NSCLC) cell activity. ASP4132 is an orally active and highly effective AMPK activator. The current study tested its activity against NSCLC cells. In primary NSCLC cells and established cell lines (A549 and NCI-H1944) ASP4132 potently inhibited cell growth, proliferation and cell cycle progression as well as cell migration and invasion. Robust apoptosis activation was detected in ASP4132-treated NSCLC cells. Furthermore, ASP4132 treatment in NSCLC cells induced programmed necrosis, causing mitochondrial p53-cyclophilin D (CyPD)-adenine nucleotide translocase 1 (ANT1) association, mitochondrial depolarization and medium lactate dehydrogenase release. In NSCLC cells ASP4132 activated AMPK signaling, induced AMPKα1-ACC phosphorylation and increased AMPK activity. Furthermore, AMPK downstream events, including mTORC1 inhibition, receptor tyrosine kinases (PDGFRα and EGFR) degradation, Akt inhibition and autophagy induction, were detected in ASP4132-treated NSCLC cells. Importantly, AMPK inactivation by AMPKα1 shRNA, knockout (using CRISPR/Cas9 strategy) or dominant negative mutation (T172A) almost reversed ASP4132-induced anti-NSCLC cell activity. Conversely, a constitutively active AMPKα1 (T172D) mimicked and abolished ASP4132-induced actions in NSCLC cells. In vivo, oral administration of a single dose of ASP4132 largely inhibited NSCLC xenograft growth in SCID mice. AMPK activation, mTORC1 inhibition and EGFR-PDGFRα degradation as well as Akt inhibition and autophagy induction were detected in ASP4132-treated NSCLC xenograft tumor tissues. Together, activation of AMPK by ASP4132 potently inhibits NSCLC cell growth in vitro and in vivo.

Highlights

Lung cancer accounts for over 13% of all new cancer diagnoses[1,2]
In ASP4132-treated pNSCLC-1 cells, G1phase cells were increased, while S-phase cells were reduced (Fig. 1D). These results indicated that the AMPK activator disrupted cell cycle progression and induced G1S arrest in pNSCLC-1 cells (Fig. 1D)
No significant apoptosis activation was detected in primary (“primary lung epithelial cells (pEpi)”) and established (BEAS2B) lung epithelial cells with the same ASP4132 treatment (Fig. 2I), further supporting cancer cell specific response by the AMPK activator. These results clearly showed that ASP4132 activated apoptosis in non-small cell lung cancer (NSCLC) cells

Summary

Introduction

Lung cancer accounts for over 13% of all new cancer diagnoses[1,2]. In the United States alone, an estimated of 228,820 adults will be diagnosed with lung cancer each year[1,2]. Of which non-small cell lung cancer (NSCLC) constitutes close to 85% of all lung cancers[1,2]. NSCLC main subtypes include adenocarcinoma, squamous cell carcinoma, and large cell carcinoma[1,2]. Adenosine monophosphate-activated protein kinase (AMPK) is a highly-conserved eukaryotic protein kinase, playing an essential role in cellular energy homeostasis[3,4,5]. With the cellular energy decreasing, activated AMPK is able to promote glucose and fatty acid uptake and oxidation[3,4,5]. AMPK is composed of α, β, and γ subunits[3,4,5]

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Results

Conclusion