LC-MS metabolomics comparisons of cancer cell and macrophage responses to methotrexate and polymer-encapsulated methotrexate

Mohammad Ahmad Al-Natour,Ali Alazzo,Amir M Ghaemmaghami,Dong-Hyun Kim,Cameron Alexander

doi:10.1016/j.ijpx.2019.100036

Abstract

Methotrexate (MTX) is a folate analogue antimetabolite widely used for the treatment of rheumatoid arthritis and cancer. A number of studies have shown that MTX delivered via nanoparticle carriers is more potent against cancer cells than free MTX, a phenomenon attributed to higher cellular uptake of the particles compared to the saturable folate receptor pathway. In this study, a cell-based global metabolic profiling approach was applied to study the effects of MTX in both free drug form and when encapsulated in -poly(lactide-co-glycolide) (PLGA) nanoparticles on a cancer cell line, A549, and also on human-like THP-1 macrophages. The results showed that MTX loaded nanoparticles had less impact on the macrophages than free MTX, and the effects on macrophages were limited to changes in nucleotide metabolism and suppression of the tricarboxylic acid cycle, whereas free MTX also led to a drop in glycolytic activity and impairment in redox homeostasis. In contrast, MTX loaded nanoparticles showed a greater impact on A549 cells than the free drug, which was in accord with studies in other cell lines in prior literature with MTX-carrier nanoparticles.

Highlights

The folate antagonist methotrexate (MTX), is used widely as an antiproliferative agent in cancer treatments and as an anti-inflammatory, for rheumatoid arthritis
Initial experiments demonstrated that the prepared NPs were well tolerated by both cells lines (Figure S1, S2), as evidenced by < 5% changes in overall metabolic activity evaluated with Alamar Blue assays
A global Liquid chromatography-mass spectrometry (LC-MS) metabolic profiling approach was employed to study the effects of MTX and nanoparticles (NPs) with entrapped MTX (Table S1) on THP-1 and A549 cells respectively

Summary

Introduction

The folate antagonist methotrexate (MTX), is used widely as an antiproliferative agent in cancer treatments and as an anti-inflammatory, for rheumatoid arthritis. Inhibition of DHFR is not enough to manage RA; MTX is assumed to have other mechanisms including inhibition of purine metabolism by inhibiting certain enzymes that are involved in that process. These include selective suppression of B cells, preventing T cell activation, downregulation of methyltransferase activity and inhibiting interleukin 1-beta binding to its receptor on the cell surface (Böhm, 2004; Brody et al, 1993; Wessels et al, 2008)

Methods

Results

Conclusion