Preclinical optimization of a gene therapy for erythromelalgia and chronic pain

摘要 慢性疼痛是持续超过正常愈合时间的疼痛。1.5亿人 全世界的老年人患有慢性疼痛，随着老年人口的增加， 糖尿病患病率上升，癌症存活率提高。慢性疼痛不仅 严重影响了许多患者的日常生活质量，也给社会经济带来了沉重的负担。 社会的负担。由于有效的治疗选择有限，慢性 尽管存在成瘾和副作用的风险，但通常用阿片类药物治疗疼痛。可惜 阿片类药物治疗慢性疼痛的处方在很大程度上助长了目前阿片类药物的流行。 因此，对于非成瘾性替代镇痛剂存在迫切且明确的未满足的需求， 慢性疼痛的治疗推动开发特定的和非成瘾性的替代品 止痛药引起了人们对一种特殊的钠通道NaV1.7的兴趣，NaV1.7被证明对 痛觉NaV1.7中的功能获得性突变与一种特征为 由四肢强烈的灼痛引起：原发性红斑性肢痛症。相反，功能丧失 NaV1.7导致感觉不到疼痛。因此，抑制NaV1.7可以是有效的 减轻疼痛和治疗红斑性肢痛症患者的方法。为了实现这一目标，我们设计了 表观遗传调节剂抑制NaV1.7的表达。而不是进行永久编辑， 在基因组中，这些表观遗传调节剂将瞬时抑制NaV1.7的表达。通过靶向 在DNA水平上，我们可以实现对NaV1.7的特异性和持久的调节， 比RNA和蛋白质靶向方法更好的药代动力学前景。本研究 我们建议优化这些表观遗传调节剂以及它们的递送，以实现高表达。 特异性和有效性。此外，我们将评估我们的优化调制器在小规模 生产研究以及大型动物模型中的毒理学研究。这样做的结果 这项研究将是一种优化的基因疗法，不仅是非成瘾性和有效的治疗 但也是高度特异性和持久性的。

ABSTRACT Chronic pain is pain that persists past the normal time of healing. 1.5 billion people worldwide suffer from chronic pain and this number continues to increase as the elderly population grows, the prevalence of diabetes rises, and cancer survival rates improve. Chronic pain not only severely impacts daily quality of life for many patients, it also places a heavy socioeconomic burden on society. Due to the limited number of efficacious treatment options available, chronic pain is often treated with opioids despite the risk of addiction and side effects. Unfortunately, the prescribing of opioids to treat chronic pain has largely fueled the current opioid epidemic. Therefore, there is an urgent and clear unmet need for non-addictive alternative analgesics for the treatment of chronic pain. The push to develop specific and non-addictive alternative painkillers has brought interest to a particular sodium channel, NaV1.7, shown to be important for pain sensing. Gain-of-function mutations in NaV1.7 are associated with a disorder characterized by intense burning pain in the extremities: primary erythromelalgia. Conversely, loss-of function of NaV1.7 results in the inability to feel pain. Therefore, inhibiting NaV1.7 can be an effective method of reducing pain and treat erythromelalgia patients. To accomplish this, we designed epigenetic modulators to repress expression of NaV1.7. Rather than making permanent edits to the genome, these epigenetic modulators will transiently inhibit expression of NaV1.7. By targeting NaV1.7 at the DNA-level, we can achieve specific and long-lasting modulation of NaV1.7, with better pharmacokinetics prospects than RNA- and protein-targeting approaches. In this study, we propose to optimize these epigenetic modulators as well as their delivery in order to achieve high specificity and efficacy. In addition, we will evaluate our optimized modulators in small-scale manufacturing studies as well as toxicological studies in a large animal model. The result of this study will be an optimized gene therapy that is not only non-addictive and efficacious for treatment of chronic pain but also highly specific and long-lasting.