Preclinical investigation of Pegylated arginase 1 as a treatment for retina and brain injury

Exp Neurol. 2022 Feb:348:113923. doi: 10.1016/j.expneurol.2021.113923. Epub 2021 Nov 12.

Abstract

Arginase 1 (A1) is the enzyme that hydrolyzes the amino acid, L-arginine, to ornithine and urea. We have previously shown that A1 deletion worsens retinal ischemic injury, suggesting a protective role of A1. In this translational study, we aimed to study the utility of systemic pegylated A1 (PEG-A1, recombinant human arginase linked to polyethylene glycol) treatment in mouse models of acute retinal and brain injury. Cohorts of WT mice were subjected to retinal ischemia-reperfusion (IR) injury, traumatic optic neuropathy (TON) or brain cerebral ischemia via middle cerebral artery occlusion (MCAO) and treated with intraperitoneal injections of PEG-A1 or vehicle (PEG only). Drug penetration into retina and brain tissues was measured by western blotting and immunolabeling for PEG. Neuroprotection was measured in a blinded fashion by quantitation of NeuN (neuronal marker) immunolabeling of retina flat-mounts and brain infarct area using triphenyl tetrazolium chloride (TTC) staining. Furthermore, ex vivo retina explants and in vitro retina neuron cultures were subjected to oxygen-glucose deprivation (OGD) followed by reoxygenation (R) and treated with PEG-A1. PEG-A1 given systemically did not cross the intact blood-retina/brain barriers in sham controls but reached the retina and brain after injury. PEG-A1 provided neuroprotection after retinal IR injury, TON and cerebral ischemia. PEG-A1 treatment was also neuroprotective in retina explants subjected to OGD/R but did not improve survival in retinal neuronal cultures exposed to OGD/R. In summary, systemic PEG-A1 administration is neuroprotective and provides an excellent route to deliver the drug to the retina and the brain after acute injury.

Keywords: Arginase; Ischemia-reperfusion injury; Neurodegeneration; Neuroprotection; Retinal ischemia; Stroke.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Arginase / pharmacokinetics
  • Arginase / therapeutic use*
  • Blood-Brain Barrier
  • Blood-Retinal Barrier
  • Brain / metabolism
  • Brain Injuries / drug therapy*
  • Brain Ischemia / drug therapy
  • Cell Survival / drug effects
  • Humans
  • Infarction, Middle Cerebral Artery / drug therapy
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Neurons / drug effects
  • Neurons / metabolism
  • Neuroprotective Agents / pharmacokinetics
  • Neuroprotective Agents / therapeutic use*
  • Optic Nerve Injuries / drug therapy
  • Polyethylene Glycols
  • Recombinant Proteins / therapeutic use
  • Reperfusion Injury / prevention & control
  • Retina / injuries*
  • Retina / metabolism

Substances

  • Neuroprotective Agents
  • Recombinant Proteins
  • Polyethylene Glycols
  • ARG1 protein, human
  • Arginase