Brain pharmacology of intrathecal antisense oligonucleotides revealed through multimodal imaging

JCI Insight. 2019 Oct 17;4(20):e129240. doi: 10.1172/jci.insight.129240.


Intrathecal (IT) delivery and pharmacology of antisense oligonucleotides (ASOs) for the CNS have been successfully developed to treat spinal muscular atrophy. However, ASO pharmacokinetic (PK) and pharmacodynamic (PD) properties remain poorly understood in the IT compartment. We applied multimodal imaging techniques to elucidate the IT PK and PD of unlabeled, radioactively labeled, or fluorescently labeled ASOs targeting ubiquitously expressed or neuron-specific RNAs. Following lumbar IT bolus injection in rats, all ASOs spread rostrally along the neuraxis, adhered to meninges, and were partially cleared to peripheral lymph nodes and kidneys. Rapid association with the pia and arterial walls preceded passage of ASOs across the glia limitans, along arterial intramural basement membranes, and along white-matter axonal bundles. Several neuronal and glial cell types accumulated ASOs over time, with evidence of probable glial accumulation preceding neuronal uptake. IT doses of anti-GluR1 and anti-Gabra1 ASOs markedly reduced the mRNA and protein levels of their respective neurotransmitter receptor protein targets by 2 weeks and anti-Gabra1 ASOs also reduced binding of the GABAA receptor PET ligand 18F-flumazenil in the brain over 4 weeks. Our multimodal imaging approaches elucidate multiple transport routes underlying the CNS distribution, clearance, and efficacy of IT-dosed ASOs.

Keywords: Neuroimaging; Neuroscience; Pharmacology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arteries / diagnostic imaging
  • Arteries / metabolism
  • Brain / blood supply
  • Brain / cytology
  • Brain / diagnostic imaging
  • Brain / metabolism*
  • Flumazenil / administration & dosage
  • Flumazenil / analogs & derivatives
  • GABA-A Receptor Antagonists / administration & dosage
  • GABA-A Receptor Antagonists / pharmacokinetics*
  • Gene Knockdown Techniques
  • Humans
  • Injections, Spinal
  • Intravital Microscopy
  • Male
  • Molecular Targeted Therapy / methods
  • Muscular Atrophy, Spinal / drug therapy*
  • Neuroglia / metabolism
  • Neurons / metabolism
  • Oligonucleotides, Antisense / administration & dosage
  • Oligonucleotides, Antisense / pharmacokinetics*
  • Pia Mater / diagnostic imaging
  • Pia Mater / metabolism
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • Rats
  • Receptors, AMPA / analysis
  • Receptors, AMPA / antagonists & inhibitors
  • Receptors, AMPA / genetics
  • Receptors, GABA-A / analysis
  • Receptors, GABA-A / genetics
  • Single Photon Emission Computed Tomography Computed Tomography
  • Spatio-Temporal Analysis
  • Thionucleotides / administration & dosage
  • Thionucleotides / pharmacokinetics
  • Tissue Distribution


  • 2'-fluoroflumazenil
  • GABA-A Receptor Antagonists
  • Gabra1 protein, rat
  • Oligonucleotides, Antisense
  • RNA, Messenger
  • Receptors, AMPA
  • Receptors, GABA-A
  • Thionucleotides
  • Flumazenil
  • glutamate receptor ionotropic, AMPA 1