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

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.

• Authors: Gerald Z Zhuang, Udita Upadhyay, Xiaoying Tong, Yuan Kang, Diana M Erasso, Eugene S Fu, Konstantinos D Sarantopoulos, Eden R Martin, Tim Wiltshire, Luda Diatchenko, Shad B Smith, William Maixner, Roy C Levitt
• Journal: Gene therapy (Gene Ther) Vol. 25 Issue 4 Pg. 297-311 (07 2018) ISSN: 1476-5462 [Electronic] England
• PMID: 29789638 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

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

Topics

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