Precision spinal gene delivery-induced functional switch in nociceptive neurons reverses neuropathic pain

Takahiro Tadokoro; Mariana Bravo-Hernandez; Kirill Agashkov; Yoshiomi Kobayashi; Oleksandr Platoshyn; Michael Navarro; Silvia Marsala; Atsushi Miyanohara; Tetsuya Yoshizumi; Michiko Shigyo; Volodymyr Krotov; Stefan Juhas; Jana Juhasova; Duong Nguyen; Helena Kupcova Skalnikova; Jan Motlik; Hana Studenovska; Vladimir Proks; Rajiv Reddy; Shawn P Driscoll; Thomas D Glenn; Taratorn Kemthong; Suchinda Malaivijitnond; Zoltan Tomori; Ivo Vanicky; Manabu Kakinohana; Samuel L Pfaff; Joseph Ciacci; Pavel Belan; Martin Marsala

doi:10.1016/j.ymthe.2022.04.023

Precision spinal gene delivery-induced functional switch in nociceptive neurons reverses neuropathic pain

Mol Ther. 2022 Aug 3;30(8):2722-2745. doi: 10.1016/j.ymthe.2022.04.023. Epub 2022 May 5.

Authors

Takahiro Tadokoro¹, Mariana Bravo-Hernandez², Kirill Agashkov³, Yoshiomi Kobayashi², Oleksandr Platoshyn², Michael Navarro², Silvia Marsala⁴, Atsushi Miyanohara⁵, Tetsuya Yoshizumi², Michiko Shigyo², Volodymyr Krotov³, Stefan Juhas⁶, Jana Juhasova⁶, Duong Nguyen⁶, Helena Kupcova Skalnikova⁶, Jan Motlik⁶, Hana Studenovska⁷, Vladimir Proks⁷, Rajiv Reddy⁸, Shawn P Driscoll⁹, Thomas D Glenn⁹, Taratorn Kemthong¹⁰, Suchinda Malaivijitnond¹⁰, Zoltan Tomori¹¹, Ivo Vanicky¹², Manabu Kakinohana¹³, Samuel L Pfaff⁹, Joseph Ciacci¹⁴, Pavel Belan¹⁵, Martin Marsala¹⁶

Affiliations

¹ Neuroregeneration Laboratory, Department of Anesthesiology, University of California, San Diego (UCSD), La Jolla, CA 92037, USA; Department of Anesthesiology, University of Ryukyus, Okinawa, Japan; Neurgain Technologies, 9620 Towne Centre Drive, Suite 100, San Diego, CA 92121, USA.
² Neuroregeneration Laboratory, Department of Anesthesiology, University of California, San Diego (UCSD), La Jolla, CA 92037, USA.
³ Departments of Sensory Signaling and Molecular Biophysics, Bogomoletz Institute of Physiology, Kyiv, Ukraine.
⁴ Neuroregeneration Laboratory, Department of Anesthesiology, University of California, San Diego (UCSD), La Jolla, CA 92037, USA; Neurgain Technologies, 9620 Towne Centre Drive, Suite 100, San Diego, CA 92121, USA.
⁵ Neuroregeneration Laboratory, Department of Anesthesiology, University of California, San Diego (UCSD), La Jolla, CA 92037, USA; Vector Core Laboratory, University of California, San Diego (UCSD), La Jolla, CA 92037, USA.
⁶ Institute of Animal Physiology and Genetics, Czech Academy of Sciences, Rumburská 89, 277 21 Liběchov, Czech Republic.
⁷ Institute of Macromolecular Chemistry, Czech Academy of Sciences, Department of Biomaterials and Bioanalogous Systems, Heyrovsky Square 2,162 06 Prague 6, Czech Republic.
⁸ Department of Anesthesiology, Pain Medicine, University of California, San Diego (UCSD), La Jolla, CA 92037, USA.
⁹ Gene Expression Laboratory and the Howard Hughes Medical Institute, Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
¹⁰ National Primate Research Center of Thailand, Chulalongkorn University, Kaengkhoi District, Saraburi 18110, Thailand.
¹¹ Department of Biophysics, Institute of Experimental Physics, Slovak Academy of Sciences, Kosice, Slovakia.
¹² Institute of Neurobiology, Biomedical Research Center, Slovak Academy of Sciences, Kosice, Slovakia.
¹³ Department of Anesthesiology, University of Ryukyus, Okinawa, Japan.
¹⁴ Department of Neurosurgery, University of California, San Diego (UCSD), La Jolla, CA 92037, USA.
¹⁵ Departments of Sensory Signaling and Molecular Biophysics, Bogomoletz Institute of Physiology, Kyiv, Ukraine; Kyiv Academic University, Kyiv, Ukraine.
¹⁶ Neuroregeneration Laboratory, Department of Anesthesiology, University of California, San Diego (UCSD), La Jolla, CA 92037, USA; Institute of Neurobiology, Biomedical Research Center, Slovak Academy of Sciences, Kosice, Slovakia. Electronic address: mmarsala@ucsd.edu.

Abstract

Second-order spinal cord excitatory neurons play a key role in spinal processing and transmission of pain signals to the brain. Exogenously induced change in developmentally imprinted excitatory neurotransmitter phenotypes of these neurons to inhibitory has not yet been achieved. Here, we use a subpial dorsal horn-targeted delivery of AAV (adeno-associated virus) vector(s) encoding GABA (gamma-aminobutyric acid) synthesizing-releasing inhibitory machinery in mice with neuropathic pain. Treated animals showed a progressive and complete reversal of neuropathic pain (tactile and brush-evoked pain behavior) that persisted for a minimum of 2.5 months post-treatment. The mechanism of this treatment effect results from the switch of excitatory to preferential inhibitory neurotransmitter phenotype in dorsal horn nociceptive neurons and a resulting increase in inhibitory activity in regional spinal circuitry after peripheral nociceptive stimulation. No detectable side effects (e.g., sedation, motor weakness, loss of normal sensation) were seen between 2 and 13 months post-treatment in naive adult mice, pigs, and non-human primates. The use of this treatment approach may represent a potent and safe treatment modality in patients suffering from spinal cord or peripheral nerve injury-induced neuropathic pain.

Keywords: GAD65 and VGAT; long-lasting pain reversal; neuropathic pain; precision spinal vector delivery; subpial injection technique; subpial non-human primates; subpial pig; targeted AAV9 delivery.

Publication types

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

MeSH terms

Animals
Gene Transfer Techniques
Mice
Neuralgia* / etiology
Neuralgia* / therapy
Nociceptors*
Posterior Horn Cells
Spinal Cord
Spinal Cord Dorsal Horn
Swine

Abstract

Publication types

MeSH terms

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