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

Gerald Z. Zhuang, Udita Upadhyay, Xiaoying Tong, Yuan Kang, Diana M. Erasso, Eugene Fu, Konstantinos D. Sarantopoulos, Eden R Martin, Tim Wiltshire, Luda Diatchenko, Shad B. Smith, William Maixner, Roy C Levitt

Research output: Contribution to journalArticle

2 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1-15
Number of pages15
JournalGene Therapy
DOIs
StateAccepted/In press - Apr 24 2018

Fingerprint

Carbonic Anhydrases
Hyperalgesia
Nerve Growth Factor
Inositol
Genetic Therapy
Analgesia
Calcium
Neuralgia
Chronic Pain
trkA Receptor
Genes
Spinal Nerves
Precision Medicine
Spinal Ganglia
Sciatic Nerve
Inbred C57BL Mouse
Virion
Ligation
Analgesics
Exons

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Human carbonic anhydrase-8 AAV8 gene therapy inhibits nerve growth factor signaling producing prolonged analgesia and anti-hyperalgesia in mice. / Zhuang, Gerald Z.; Upadhyay, Udita; Tong, Xiaoying; Kang, Yuan; Erasso, Diana M.; Fu, Eugene; Sarantopoulos, Konstantinos D.; Martin, Eden R; Wiltshire, Tim; Diatchenko, Luda; Smith, Shad B.; Maixner, William; Levitt, Roy C.

In: Gene Therapy, 24.04.2018, p. 1-15.

Research output: Contribution to journalArticle

Zhuang, Gerald Z. ; Upadhyay, Udita ; Tong, Xiaoying ; Kang, Yuan ; Erasso, Diana M. ; Fu, Eugene ; Sarantopoulos, Konstantinos D. ; Martin, Eden R ; Wiltshire, Tim ; Diatchenko, Luda ; Smith, Shad B. ; Maixner, William ; Levitt, Roy C. / Human carbonic anhydrase-8 AAV8 gene therapy inhibits nerve growth factor signaling producing prolonged analgesia and anti-hyperalgesia in mice. In: Gene Therapy. 2018 ; pp. 1-15.
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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.",
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AU - Kang, Yuan

AU - Erasso, Diana M.

AU - Fu, Eugene

AU - Sarantopoulos, Konstantinos D.

AU - Martin, Eden R

AU - Wiltshire, Tim

AU - Diatchenko, Luda

AU - Smith, Shad B.

AU - Maixner, William

AU - Levitt, Roy C

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