Human carbonic anhydrase-8 AAV8 gene therapy inhibits nerve growth factor signaling producing prolonged analgesia and anti-hyperalgesia in mice

Gene Ther. 2018 Jul;25(4):297-311. doi: 10.1038/s41434-018-0018-7. Epub 2018 Apr 24.

Abstract

Carbonic anhydrase-8 (Car8; murine gene symbol) is an allosteric inhibitor of inositol trisphosphate receptor-1 (ITPR1), which regulates neuronal intracellular calcium release. We previously reported that wild-type Car8 overexpression corrects the baseline allodynia and hyperalgesia associated with calcium dysregulation in the waddle (wdl) mouse due to a 19 bp deletion in exon 8 of the Car8 gene. In this report, we provide preliminary evidence that overexpression of the human wild-type ortholog of Car8 (CA8WT), but not the reported CA8 S100P loss-of-function mutation (CA8MT), inhibits nerve growth factor (NGF)-induced phosphorylation of ITPR1, TrkA (NGF high-affinity receptor), and ITPR1-mediated cytoplasmic free calcium release in vitro. In addition, we show that gene transfer using AAV8-V5-CA8WT viral particles via sciatic nerve injection demonstrates retrograde transport to dorsal root ganglia (DRG) producing prolonged V5-CA8WT expression, pITPR1 and pTrkA inhibition, and profound analgesia and anti-hyperalgesia in male C57BL/6J mice. AAV8-V5-CA8WT-mediated overexpression prevented and treated allodynia and hyperalgesia associated with chronic neuropathic pain produced by the spinal nerve ligation (SNL) model. These AAV8-V5-CA8 data provide a proof-of-concept for precision medicine through targeted gene therapy of NGF-responsive somatosensory neurons as a long-acting local analgesic able to prevent and treat chronic neuropathic pain through regulating TrkA signaling, ITPR1 activation, and intracellular free calcium release by ITPR1.

Publication types

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

MeSH terms

  • Analgesia / methods
  • Animals
  • Biomarkers, Tumor / biosynthesis
  • Biomarkers, Tumor / genetics*
  • Dependovirus / genetics
  • Disease Models, Animal
  • Genetic Therapy / methods*
  • Humans
  • Hyperalgesia / genetics
  • Hyperalgesia / therapy*
  • Inositol 1,4,5-Trisphosphate Receptors / drug effects*
  • Inositol 1,4,5-Trisphosphate Receptors / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Nerve Growth Factor / antagonists & inhibitors*
  • Nerve Growth Factor / genetics
  • Nerve Growth Factor / metabolism
  • Neuralgia / genetics
  • Neuralgia / therapy
  • Neurons / metabolism
  • Pain Management / methods
  • Phosphorylation
  • Signal Transduction

Substances

  • Biomarkers, Tumor
  • CA8 protein, human
  • Inositol 1,4,5-Trisphosphate Receptors
  • NGF protein, human
  • Nerve Growth Factor