The Multikinase Inhibitor AD80 Induces Mitotic Catastrophe and Autophagy in Pancreatic Cancer Cells

Abstract

Significant advances in understanding the molecular complexity of the development and progression of pancreatic cancer have been made, but this disease is still considered one of the most lethal human cancers and needs new therapeutic options. In the present study, the antineoplastic effects of AD80, a multikinase inhibitor, were investigated in models of pancreatic cancer. AD80 reduced cell viability and clonogenicity and induced polyploidy in pancreatic cancer cells. At the molecular level, AD80 reduced RPS6 and histone H3 phosphorylation and induced γH2AX and PARP1 cleavage. Additionally, the drug markedly decreased AURKA phosphorylation and expression. In PANC-1 cells, AD80 strongly induced autophagic flux (consumption of LC3B and SQSTM1/p62). AD80 modulated 32 out of 84 autophagy-related genes and was associated with vacuole organization, macroautophagy, response to starvation, cellular response to nitrogen levels, and cellular response to extracellular stimulus. In 3D pancreatic cancer models, AD80 also effectively reduced growth independent of anchorage and cell viability. In summary, AD80 induces mitotic aberrations, DNA damage, autophagy, and apoptosis in pancreatic cancer cells. Our exploratory study establishes novel targets underlying the antineoplastic activity of the drug and provides insights into the development of therapeutic strategies for this disease.

Oxidative stress induces release of mitochondrial DNA into the extracellular space in human placental villous trophoblast BeWo cells

“…Proteins were transferred for 90 minutes to nitrocellulose or polyvinylidene difluoride (PVDF) membranes. Afterwards, membranes were blocked for 1 hour with 3% Bovine Serum Albumin or with 5% non-fat dry milk (for LC3 A/B) as described by Lima et al (23) Primary antibodies were dissolved with 3% BSA in tris buffered saline (TBST) and membranes were incubated overnight at 4°C with SQSTM1/p62 (Cell Signaling, cat# 88588, 1:1000). The primary LC3 A/B (Cell Signaling, cat# 4108s, 1:1000) was dissolved with 3% non-fat dry milk.…”

Modulating in vitro lung fibroblast activation via senolysis of senescent human alveolar epithelial cells

“…To investigate novel mechanisms of existing late-stage clinical assets, AD80, a multi-kinase inhibitor targeting RET (c-RET), BRAF, S6K, and SRC that is under investigation for the treatment of certain cancers, was included as a potential senolytic. AD80 has shown apoptosis-inducing effects in various cell types, particularly on cells in G 2 /M phase [ 31 , 32 ], and was hypothesized to also impact injured AECs. ABT-317, a JAK1-selective inhibitor, was also tested as a potential senomorphic that is more selective than ruxolitinib [ 33 ].…”