Extracellular vesicles as dynamic surrogates for drug transporter expression. A new tool to personalize pharmacotherapy?

• Applicant: Dr. Juan Pablo Rigalli
• Subject Area: Pharmacology ; Cell Biology
• Term: from 2022 to 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 509856975

• Final Report Year: 2023

Final Report Abstract

The P-glycoprotein (P-gp), the multidrug resistance-associated protein 2 (MRP2) and the breast cancer resistance protein (BCRP) belong to the superfamily of ABC transporters (ATP binding cassette) and play a major role in drug pharmacokinetics. They are expressed in tissues of high pharmacological relevance such as the liver, the kidneys, the intestine and the blood-brain barrier. Also, they account for the resistance towards chemotherapeutic agents in cancer cells. The expression of ABC transporters is highly variable within the population and can be regulated by endo- and xenobiotics. Higher transporter expression may lead to higher drug efflux and therapy failure. Lower transporter expression may lead to lower drug efflux and toxicity. Therefore, the individualized estimation of the expression of ABC transporters could aid to adjust the therapy to avoid cases of over- and under-exposure. So far, repeated extraction of biopsies would be the only option to measure ABC transporter expression in relevant tissues. However, due to its invasive character, this cannot be performed routinely. Extracellular vesicles (EVs) are nanoparticles released by all cell types of the organism. Their cargo consists of proteins, RNA, and lipids, the array of these components being a fingerprint of the cell of origin. The aim of this project was to investigate the potential of the ABC transportercargo in EVs as a dynamic surrogate for the transporter expression in normal and tumoral cells and thus, for their drug excretion capacity. We validated an ultrasensitive UPLC-MS/MS assay for the simultaneous quantification of P-gp, MRP2 and BCRP in EVs and the cells of origin. Treatment of MCF7 (breast cancer) and HepG2 (hepatocellular carcinoma) cells with rifampicin and, rifampicin and hypericin resulted in BCRP up-regulation and MRP2 down-regulation (expression and activity level) in the cells, respectively. Only in HepG2 cells, these changes in the protein expression were fully reproduced by EVs, albeit the onset of the effect was observed at a later time-point. This may be attributed to the time required for the EV biogenesis. Altogether, the use of EVs allows to estimate changes in the expression of MRP2 in one hepatocellular carcinoma cell line. If this is confirmed in vivo, EV analysis could allow to stratify the patients based on their drug clearance capacity and, thus, adjust pharmacotherapy and, ultimately, prevent situations of toxicity or therapy failure.

Publications

• Potential of extracellular vesicles as surrogates for the cellular ABC transporter expression. Annual Meeting of the Italian Society of Clinical Pharmacy and Therapy. Rome, November 24-26th, 2022.
  Gagliardi A.; Rigalli JP, Weiss J.; Sauter M.; Barocelli E.; Borghesi S. & Gazzola A. M.
  
• Extracellular Vesicles and Cancer Multidrug Resistance: Undesirable Intercellular Messengers?. Life, 13(8), 1633.
  Bucci-Muñoz, María; Gola, Aldana Magalí; Rigalli, Juan Pablo; Ceballos, María Paula & Ruiz, María Laura
  
• Extracellular Vesicles as Surrogates for Drug Metabolism and Clearance: Promise vs. Reality. Life, 13(8), 1745.
  Gagliardi, Anna; Bajraktari-Sylejmani, Gzona; Barocelli, Elisabetta; Weiss, Johanna & Rigalli, Juan Pablo
  
• Liquid biopsy and extracellular vesicles (EVs). A new horizon in the precision oncology landscape. International Student Congress of Clinical Innovation and Medical Sciences. Parma (Italy), November 9-11th, 2023.
  Gagliardi A.; Rigalli J. P.; Weiss J. & Barocelli E.