Gene knockdown with lentiviral vector‐mediated intrathecal RNA interference of protein kinase C gamma reverses chronic morphine tolerance in rats

Song, Zongbin; Zou, Wangyuan; Liu, Chang; Guo, Qulian

doi:10.1002/jgm.1514

Cited by 28 publications

(25 citation statements)

References 48 publications

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“…without using viral vectors (29), to provide gene knockdown or gene silencing using shRNA or siRNA (30,31), as well as in regeneration strategies using neural precursor cells (32). However, none of those previous studies focused specifically on myelinating Schwann cells, because ubiquitous promoters were used.…”

Section: Discussionmentioning

confidence: 99%

Intrathecal gene therapy rescues a model of demyelinating peripheral neuropathy

Kagiava

Sargiannidou

Theophilidis

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Inherited demyelinating peripheral neuropathies are progressive incurable diseases without effective treatment. To develop a gene therapy approach targeting myelinating Schwann cells that can be translatable, we delivered a lentiviral vector using a single lumbar intrathecal injection and a myelin-specific promoter. The human gene of interest, GJB1, which is mutated in X-linked CharcotMarie-Tooth Disease (CMT1X), was delivered intrathecally into adult Gjb1-null mice, a genetically authentic model of CMT1X that develops a demyelinating peripheral neuropathy. We obtained widespread, stable, and cell-specific expression of connexin32 in up to 50% of Schwann cells in multiple lumbar spinal roots and peripheral nerves. Behavioral and electrophysiological analysis revealed significantly improved motor performance, quadriceps muscle contractility, and sciatic nerve conduction velocities. Furthermore, treated mice exhibited reduced numbers of demyelinated and remyelinated fibers and fewer inflammatory cells in lumbar motor roots, as well as in the femoral motor and sciatic nerves. This study demonstrates that a single intrathecal lentiviral gene delivery can lead to Schwann cell-specific expression in spinal roots extending to multiple peripheral nerves. This clinically relevant approach improves the phenotype of an inherited neuropathy mouse model and provides proof of principle for treating inherited demyelinating neuropathies.demyelinating neuropathy | Charcot-Marie-Tooth disease | connexin32 | peripheral nerve | gene therapy

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Section: Discussionmentioning

confidence: 99%

Intrathecal gene therapy rescues a model of demyelinating peripheral neuropathy

Kagiava

Sargiannidou

Theophilidis

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…To induce morphine tolerance, 10 μg morphine sulfate (in a volume of 10 μL) was delivered via an intrathecal catheter twice daily for 7 d. The control group was injected with 10 μL normal saline. Injections were followed by administration of 10 μL normal saline to flush the catheter [52]. Morphine analgesia was assessed on the test day using the tail-flick test both before and 30 min after morphine administration.…”

Section: Methodsmentioning

confidence: 99%

miR-219-5p targets CaMKIIγ to attenuate morphine tolerance in rats

Wang

Shao

et al. 2017

Oncotarget

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Morphine tolerance is a clinical challenge in pain management. Emerging evidence suggests that microRNA (miRNA) plays a regulatory role in the development of morphine tolerance. miR-219-5p (miR-219) targets calmodulin-dependent protein kinase II γ (CaMKIIγ) to activate central pain sensitization via N-methyl-D-aspartate (NMDA) receptor. Therefore, we hypothesized that miR-219-5p attenuates morphine tolerance by targeting CaMKIIγ. We found that the expression of miR-219-5p was decreased significantly after chronic morphine treatment. Overexpression of miR-219-5p by lentivirus injection prevents the development of morphine tolerance. CaMKIIγ, the target gene of miR-219-5p was downregulated by overexpression of miR-219-5p both in vivo and in vitro. Furthermore, we found that lentiviral-mediated miR-219-5p decreased the expression of NMDA receptor subunit 1 (NR1), leading to attenuation of morphine tolerance. Overall, the data demonstrate that miR-219-5p plays a crucial role in alleviating morphine tolerance by inhibiting the CaMKII/NMDA receptor pathway. Overexpression of miR-219-5p may be a potential strategy to ameliorate morphine tolerance.

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“…Accordingly, a variety of approaches have been explored to elucidate the potential role of PKCγ as a pain mediator in nociceptive signal transduction [26-29]. It has been demonstrated that PKCγ is involved in many aspects of cellular sensitization, including modulation of channel conductivity, increased trafficking of receptors, and release of excitatory neurotransmitters [30].…”

Section: Introductionmentioning

confidence: 99%

PKCγ Receptor Mediates Visceral Nociception and Hyperalgesia following Exposure to PTSD-Like Stress in the Spinal Cord of Rats

Chen

et al. 2013

Mol Pain

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BackgroundClinical studies indicate that patients with post-traumatic stress disorder (PTSD) frequently share comorbidity with numerous chronic pain conditions. However, the sustained effects of PTSD-like stress over time on visceral nociception and hyperalgesia have been rarely studied, and the underlying mechanisms of stress-induced modulation of visceral hyperalgesia remain elusive. The purpose of this study was to investigate the characterization of visceral nociception and hyperalgesia over time in rats exposed to PTSD-like stress, and to explore the potential role of protein kinase C gamma (PKCγ) in mediating visceral hyperalgesia following exposure to PTSD-like stress.ResultsOn day 1, the rats exposed to single-prolonged stress (SPS, an established animal model for PTSD) exhibited an analgesic response and its visceromotor response (VMR) to graded colorectal distention (CRD) at 40 and 60 mmHg was reduced compared with the control group (all P < 0.05). On day 6, the VMR returned to the baseline value. However, as early as 7 days after SPS, VMR dramatically increased compared with its baseline value and that in the controls (all P < 0.001) and this increase persisted for 28 days, with the peak on day 9. Abdominal withdrawal reflex (AWR) scores were higher in SPS rats than in controls on days 7, 9, 14, 21 and 28 (all P < 0.001). Intrathecal administration of GF109203X (an inhibitor of PKC gamma), attenuated the SPS-induced increase in both VMR and AWR scores on days 7, 14, 21 and 28 (all P < 0.05). PKCγ protein expression determined by immunofluorescence was reduced in the spinal cord within 3 days after the exposure to SPS (P < 0.01), which returned to normal levels between days 4 and 6, and significantly increased from day 7, and this increase was maintained on days 14, 21, and 28 (all P < 0.001), with the peak on day 9. In addition, Western blotting showed a consistent trend in the changes of PKCγ protein expression.ConclusionsThe modified SPS alters visceral sensitivity to CRD, and contributes to the maintenance of visceral hyperalgesia, which is associated with enhanced PKCγ expression in the spinal cord. Functional blockade of the PKCγ receptors attenuates SPS-induced visceral hyperalgesia. Thus, the present study identifies a specific molecular mechanism for visceral hyperalgesia which may pave the way for novel therapeutic strategies for PTSD-like conditions.

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Gene knockdown with lentiviral vector‐mediated intrathecal RNA interference of protein kinase C gamma reverses chronic morphine tolerance in rats

Abstract: A single injection of LV-shPKCγ reversed MT by reducing the expression of PKCγ in the spinal cord. These findings indicate that the use of LV-shPKCγ might be a potential strategy for therapy in MT.

Cited by 28 publications

References 48 publications

Intrathecal gene therapy rescues a model of demyelinating peripheral neuropathy

Intrathecal gene therapy rescues a model of demyelinating peripheral neuropathy

miR-219-5p targets CaMKIIγ to attenuate morphine tolerance in rats

PKCγ Receptor Mediates Visceral Nociception and Hyperalgesia following Exposure to PTSD-Like Stress in the Spinal Cord of Rats

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