Magnetic Resonance and Ultrastructural Characterization of PEGylation-associated Vacuolation in Nonclinical Models

Toxicol Pathol. 2017 Jul;45(5):604-613. doi: 10.1177/0192623317714068. Epub 2017 Jul 4.

Abstract

Conjugation with polyethylene glycol (PEG) is a strategy for improving the pharmaceutical properties of therapeutic proteins. In nonclinical studies of PEGylated compounds, microscopic tissue vacuolation is often observed, characterized ultrastructurally in this report by lysosomal distension. Although PEGylation-associated vacuolation appears to be of limited toxicologic concern when alternative therapies are limited, the risk-benefit considerations may be impacted by uncertainty about reversibility, lack of methods for monitoring PEG accumulation in vivo without biopsy, and the variability in tissues affected depending on species studied. We demonstrate the use of magnetic resonance spectroscopy (MRS) to measure PEG concentrations at multiple time points in vivo in the kidney with comparison to PEG concentrations ex vivo in body fluids and tissue extracts using nuclear magnetic resonance (NMR) spectroscopy. Furthermore, we demonstrate the use of these techniques to study distribution and elimination of PEG in a dog model of PEGylation-associated vacuolation. This report suggests that MRS could be further investigated as a feasible imaging-based method for monitoring PEG accumulation in a clinical setting in conjunction with NMR quantitation of PEG in plasma and urine.

Keywords: PEGylation; biologics; dog; magnetic resonance; pharmaceutical; protein therapeutics; rat; vacuolation.

MeSH terms

  • Animals
  • Cerebral Cortex / chemistry
  • Cerebral Cortex / cytology
  • Cerebral Cortex / metabolism
  • Dogs
  • Female
  • Kidney / chemistry
  • Kidney / cytology
  • Kidney / metabolism
  • Magnetic Resonance Spectroscopy / methods*
  • Male
  • Polyethylene Glycols / chemistry*
  • Polyethylene Glycols / metabolism*
  • Polyethylene Glycols / pharmacokinetics
  • Proteins / chemistry*
  • Proteins / metabolism*
  • Proteins / pharmacokinetics
  • Rats
  • Rats, Sprague-Dawley
  • Spleen / chemistry
  • Spleen / cytology
  • Spleen / metabolism
  • Tissue Distribution
  • Vacuoles / chemistry*
  • Vacuoles / metabolism
  • Vacuoles / ultrastructure

Substances

  • Proteins
  • Polyethylene Glycols