Next Generation of Cancer Drug Repurposing: Therapeutic Combination of Aspirin and Oseltamivir Phosphate Potentiates Gemcitabine to Disable Key Survival Pathways Critical for Pancreatic Cancer Progression

Abstract

Resistance to chemotherapeutics and high metastatic rates contribute to the abysmal survival rate in patients with pancreatic cancer. An alternate approach for treating human pancreatic cancer involves repurposing the anti-inflammatory drug, aspirin (ASA), with oseltamivir phosphate (OP) in combination with the standard chemotherapeutic agent, gemcitabine (GEM). The question is whether treatment with ASA and OP can sensitize cancer cells to the cytotoxicity induced by GEM and limit the development of chemoresistance. To assess the key survival pathways critical for pancreatic cancer progression, we used the AlamarBlue cytotoxicity assay to determine the cell viability and combination index for the drug combinations, flow cytometric analysis of annexin V apoptosis assay to detect apoptotic and necrotic cells, fluorometric QCM™ chemotaxis migration assay to assess cellular migration, fluorometric extracellular matrix (ECM) cell adhesion array kit to assess the expression of the ECM proteins, scratch wound assay using the 96-well WoundMaker™, and the methylcellulose clonogenic assay to assess clonogenic potential. The combination of ASA and OP with GEM significantly upended MiaPaCa-2 and PANC-1 pancreatic cancer cell viability, clonogenic potential, expression of critical extracellular matrix proteins, migration, and promoted apoptosis. ASA in combination with OP significantly improves the effectiveness of GEM in the treatment of pancreatic cancer and disables key survival pathways critical to disease progression.

Repositioning of Old Drugs for Novel Cancer Therapies: Continuous Therapeutic Perfusion of Aspirin and Oseltamivir Phosphate with Gemcitabine Treatment Disables Tumor Progression, Chemoresistance, and Metastases

“…Since we established that the continuous perfusion of ASA + OP in an osmotic pump presents a promising option for treating pancreatic cancer, we questioned whether ASA, OP, or ASA + OP loaded pumps would be effective treatment options compared to ASA + OP injections (INJ) for the treatment of pancreatic cancer. The rationale is that the combination of ASA and OP with GEM treatment significantly upends MiaPaCa-2 and PANC-1 pancreatic cancer cell viability, clonogenicity potential, and expression of critical extracellular matrix expression proteins, migration, and induces apoptosis, as previously reported by us [ 6 ].…”

“…We have previously reported using the methylcellulose clonogenicity assay on MiaPaCa-2 and PANC-1 cells to determine metastatic, resistant pancreatic progenitor cells to proliferate and differentiate into colonies using in a semi-solid media [ 6 ]. PANC-1 cells revealed a statistically significant decreased in the clonogenicity potential following treatment with ASA + GEM, OP + GEM, and ASA + OP + GEM compared to the CTRL and GEM alone ( p < 0.01) [ 6 ].…”

“…We have previously reported using the methylcellulose clonogenicity assay on MiaPaCa-2 and PANC-1 cells to determine metastatic, resistant pancreatic progenitor cells to proliferate and differentiate into colonies using in a semi-solid media [ 6 ]. PANC-1 cells revealed a statistically significant decreased in the clonogenicity potential following treatment with ASA + GEM, OP + GEM, and ASA + OP + GEM compared to the CTRL and GEM alone ( p < 0.01) [ 6 ]. Here, the xenograft tumors at necropsy were fixed, paraffin-embedded, and stained with antibodies for pancreatic cell differentiation marker PDX-1, proliferation marker Ki67, and the TUNEL assay to measure apoptotic cells in the tumor ( Figure 4 ).…”

“…Intrinsic resistance of pancreatic cancer cells to chemotherapy treatment is well recognized clinically and, in the laboratory [ 5 , 6 , 7 , 8 , 9 ]. However, combining first-line chemotherapies in treating metastatic pancreatic cancer, including Abraxane with gemcitabine (GEM) or 5-FU with Oxaliplatin and Irinotecan, can control the disease process for a median length of time of approximately six months in patients studied [ 3 , 4 , 10 , 11 ].…”

“…We also incorporated ASA based on our previous work showing that ASA specifically targeted and inhibited Neu-1 sialidase activity [ 7 ]. Notably, Neu1 regulates EGF-induced receptor activation in pancreatic PANC-1 cancer cells and its GEM-resistant variant, PANC-1-GemR cells [ 6 , 7 ].…”