CRISPR/Cas9-Based Mutagenesis of Histone H3.1 in Spinal Dynorphinergic Neurons Attenuates Thermal Sensitivity in Mice

Abstract

Burn injury is a trauma resulting in tissue degradation and severe pain, which is processed first by neuronal circuits in the spinal dorsal horn. We have recently shown that in mice, excitatory dynorphinergic (Pdyn) neurons play a pivotal role in the response to burn-injury-associated tissue damage via histone H3.1 phosphorylation-dependent signaling. As Pdyn neurons were mostly associated with mechanical allodynia, their involvement in thermonociception had to be further elucidated. Using a custom-made AAV9_mutH3.1 virus combined with the CRISPR/cas9 system, here we provide evidence that blocking histone H3.1 phosphorylation at position serine 10 (S10) in spinal Pdyn neurons significantly increases the thermal nociceptive threshold in mice. In contrast, neither mechanosensation nor acute chemonociception was affected by the transgenic manipulation of histone H3.1. These results suggest that blocking rapid epigenetic tagging of S10H3 in spinal Pdyn neurons alters acute thermosensation and thus explains the involvement of Pdyn cells in the immediate response to burn-injury-associated tissue damage.

Elevated 18:0 lysophosphatidylcholine contributes to the development of pain in tissue injury

“…4B). 52,85 While pretreatment of cells with 1 mM 18:0 LPC did not increase the overall number of heat-responding neurons (142 out of 441 (32.2%), Fischer exact test, P 5 0.78), it did change the distribution of heat thresholds; they formed 3 rather than 4 groups (Fig. 4C; 36, 42 and 48.6˚C, with a cumulative threshold of 41.9˚C).…”

“… 75 , 79 Furthermore, blocking phosphorylation of S10 in H3 in spinal cord dynorphinergic neurons, which constitute the majority of neurons exhibiting up-regulation in p-S10H3 after burn injury, 84 significantly reduces responsiveness of mice to noxious heat. 52 Importantly, although chlorpromazine is an antagonist/inverse agonist of a series of dopamine, serotonin, and α2 adrenergic receptors, which are expressed in various components of spinal dorsal horn circuitries and supraspinal neurons that send fibres into, and modulate nociceptive processing in, the spinal dorsal horn, it has no analgesic effect in tissue injury. 6 , 9 , 26 , 28 , 88 Therefore, the only plausible explanation for the effect of chlorpromazine on p-S10H3 expression is that, following chlorpromazine injection, the spinal nociceptive input from primary sensory neurons was reduced.…”

Pathology of pain and its implications for therapeutic interventions

“… 303 Remolding phosphorylation sites on histone H3.1 by AAV raises the threshold for thermal nociception. 703 The indirect regulatory functions of gene therapies may offer more rapid responses and enhanced safety based on the features of epigenetics and PTMs. Furthermore, introduction of the ASC-encoding gene effectively suppresses the schwannoma growth and mitigates associated cancer pain perception.…”