101. Human carbonic anhydrase-8 AAV8 gene therapy inhibits nerve growth factor signaling producing prolonged analgesia and anti-hyperalgesia in mice.
- Author
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Zhuang GZ, Upadhyay U, Tong X, Kang Y, Erasso DM, Fu ES, Sarantopoulos KD, Martin ER, Wiltshire T, Diatchenko L, Smith SB, Maixner W, and Levitt RC
- Subjects
- Analgesia methods, Animals, Biomarkers, Tumor biosynthesis, Dependovirus genetics, Disease Models, Animal, Humans, Hyperalgesia genetics, Inositol 1,4,5-Trisphosphate Receptors metabolism, Male, Mice, Mice, Inbred C57BL, Nerve Growth Factor genetics, Nerve Growth Factor metabolism, Neuralgia genetics, Neuralgia therapy, Neurons metabolism, Pain Management methods, Phosphorylation, Signal Transduction, Biomarkers, Tumor genetics, Genetic Therapy methods, Hyperalgesia therapy, Inositol 1,4,5-Trisphosphate Receptors drug effects, Nerve Growth Factor antagonists & inhibitors
- 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 (CA8
WT ), 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.- Published
- 2018
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