CRISPR Epigenome Editing of AKAP150 in DRG Neurons Abolishes Degenerative IVD-Induced Neuronal Activation.
CRISPR Epigenome Editing of AKAP150 in DRG Neurons Abolishes Degenerative IVD-Induced Neuronal Activation.
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DOI：
10.1016/j.ymthe.2017.06.010
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发表时间：
2017-09
期刊：
影响因子：
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通讯作者：
J. Stover;Niloofar Farhang;Kristofer C. Berrett;Jason Gertz;B. Lawrence;R. Bowles
中科院分区：
文献类型：
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作者：
J. Stover;Niloofar Farhang;Kristofer C. Berrett;Jason Gertz;B. Lawrence;R. Bowles
背痛是残疾的主要原因，在世界范围内具有重大的社会经济影响。退行性椎间盘（IVD）已被假设为导致背痛，但需要更好地了解退行性IVD和支配椎间盘的伤害性神经元之间的相互作用以及直接针对这些相互作用的治疗策略，以提高我们对背痛的理解和治疗。我们研究了退行性IVD诱导的背根神经节（DRG）神经元活性的变化，并利用CRISPR表观基因组编辑作为神经调节策略。通过将DRG神经元暴露于正常和病理IVD pH水平下的退行性IVD条件培养基，我们证明退行性IVD触发白细胞介素（IL）-6诱导的神经元活性对热刺激的增加，这是由AKAP直接介导的，并通过酸性pH增强。我们开发了慢病毒CRISPR表观基因组编辑载体，其通过靶向启动子组蛋白甲基化调节AKAP 150的内源性表达。当递送至DRG神经元时，这些表观基因组修饰载体消除了退行性IVD诱导的DRG升高的神经元活性，同时保留了非病理性神经元活性。这项工作阐明了CRISPR表观基因组编辑作为基于靶基因的疼痛神经调节策略的潜力。
Back pain is a major contributor to disability and has significant socioeconomic impacts worldwide. The degenerative intervertebral disc (IVD) has been hypothesized to contribute to back pain, but a better understanding of the interactions between the degenerative IVD and nociceptive neurons innervating the disc and treatment strategies that directly target these interactions is needed to improve our understanding and treatment of back pain. We investigated degenerative IVD-induced changes to dorsal root ganglion (DRG) neuron activity and utilized CRISPR epigenome editing as a neuromodulation strategy. By exposing DRG neurons to degenerative IVD-conditioned media under both normal and pathological IVD pH levels, we demonstrate that degenerative IVDs trigger interleukin (IL)-6-induced increases in neuron activity to thermal stimuli, which is directly mediated by AKAP and enhanced by acidic pH. Utilizing this novel information on AKAP-mediated increases in nociceptive neuron activity, we developed lentiviral CRISPR epigenome editing vectors that modulate endogenous expression of AKAP150 by targeted promoter histone methylation. When delivered to DRG neurons, these epigenome-modifying vectors abolished degenerative IVD-induced DRG-elevated neuron activity while preserving non-pathologic neuron activity. This work elucidates the potential for CRISPR epigenome editing as a targeted gene-based pain neuromodulation strategy.