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.
Gerald Z Zhuang  1 ,  Udita Upadhyay  1 ,  Xiaoying Tong  1 ,  Yuan Kang  1 ,  Diana M Erasso  1 ,  Eugene S Fu  1 ,  Konstantinos D Sarantopoulos  1  2 ,  Eden R Martin  3  4 ,  Tim Wiltshire  5 ,  Luda Diatchenko  6 ,  Shad B Smith  7 ,  William Maixner  7 ,  Roy C Levitt  8  9  10  11
Affiliations
  • 1. Department of Anesthesiology, Perioperative Medicine and Pain Management, University of Miami Miller School of Medicine, Miami, FL, USA.
  • 2. Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
  • 3. John T. MacDonald Foundation, Department of Human Genetics, University of Miami Miller School of Medicine, Miami, FL, USA.
  • 4. John P. Hussman Institute for Human Genomics, University of Miami Miller School of Medicine, Miami, FL, USA.
  • 5. Department of Pharmacology and Experimental Therapeutics, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC, USA.
  • 6. Alan Edwards Centre for Research on Pain, Department of Anesthesiology, McGill University, Montreal, Quebec, Canada.
  • 7. Center for Translational Pain Medicine, Department of Anesthesiology, Duke University School of Medicine, Durham, NC, USA.
  • 8. Department of Anesthesiology, Perioperative Medicine and Pain Management, University of Miami Miller School of Medicine, Miami, FL, USA. [email protected].
  • 9. Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, USA. [email protected].
  • 10. John T. MacDonald Foundation, Department of Human Genetics, University of Miami Miller School of Medicine, Miami, FL, USA. [email protected].
  • 11. John P. Hussman Institute for Human Genomics, University of Miami Miller School of Medicine, Miami, FL, USA. [email protected].
PMID: 29789638 DOI: 10.1038/s41434-018-0018-7
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.

Products