Low Electric Field-Based Gene Delivery of IL-30 Gene Therapy for Liver Injury

 描述：肝损伤，如肝纤维化和肝硬变，常直接或间接导致死亡。传统的治疗方法--通常是小分子药物--无法治愈严重的肝脏损伤，尽管这些药物对患者有很大的好处，但一些药物对肝脏也有毒性。因此，迫切需要开发有效的、无毒的治疗方法来预防和/或减少肝脏损伤。在目前的应用中，我们建议研究一种非传统的治疗肝损伤的方法，即非病毒、低电场电穿孔传递白介素30(IL-30)基因治疗。我们将使用IL-30，因为我们之前的研究表明，它在健康人的肝脏中高表达，而在肝硬变的肝脏中低表达(参见附录I中的肝病学中的酒吧)，这表明这种细胞因子是一种天然的生理肝脏保护剂。事实上，我们的初步结果表明，IL-30可以抑制炎症诱导的小鼠模型的肝损伤，可能是通过导致激活的星状细胞凋亡，从而减少炎症细胞因子和纤维化形成胶原的产生来源。我们将使用电穿孔来提供IL-30基因治疗，因为我们和其他人已经发现，电穿孔是将DNA输送到肌肉和皮肤的一种非常有效的方法。此外，临床试验表明，电穿孔可以安全地输送疫苗和抗癌基因治疗。因此，电穿孔介导的IL-30基因治疗具有很大的临床应用潜力。IL-30‘S作用的生物学机制的研究为今后IL-30基因治疗与其他途径相结合成功治疗肝病奠定了基础。我们的中心假设是，低电场电穿孔传递的IL-30基因治疗在不引起肝脏毒性的情况下减少和修复肝损伤，这种肝损伤的减轻和修复是通过IL-30‘S激活NKG2D配体-NKG2D受体介导的纤维化前体细胞的死亡来实现的。为了验证我们的假设，我们将追求以下两个具体目标。目标1：利用以皮肤为基础的低成本 电场介导型IL-30基因治疗慢性肝损伤目的：探讨IL-30基因治疗减轻肝损伤的生物学机制。这些目标的成功实现将为基因治疗提供一种简单、实用、潜在有效的方法；使我们能够将IL-30基因治疗转化为未来的临床试验；并揭示IL-30抑制肝损伤的新机制。这种新的治疗方法可能是对标准治疗的补充，以修复目前被认为是无法治愈的肝损伤。

 DESCRIPTION: Liver injury such as fibrosis and cirrhosis often directly or indirectly causes death. Conventional treatments- usually small-molecule drugs-do not cure severe liver injury, and although these agents provide great benefit in patients, some are also toxic to the liver. Therefore, there is an urgent need to develop effective, nontoxic therapies that prevent and/or reduce liver injury. In the current application, we propose investigating an unconventional approach to treating liver injury, namely, non-viral, low-electric-field electroporation delivery o interleukin-30 (IL-30) gene therapy. We will use IL-30 because our previous studies revealed that its expression is high in healthy human livers but low in cirrhotic livers (see the pub in Hepatology in Appendix I), suggesting that this cytokine is a natural and physiological hepatoprotectant. Indeed, our preliminary results show that IL-30 can inhibit inflammation-induced liver injury in a murine model by possibly causing the apoptosis of the activated stellate cells, which reduces the production source of inflammatory cytokines and fibrosis-forming collagen. We will use electroporation to deliver IL-30 gene therapy because we and others have found that electroporation is a very effective method of delivering DNA into muscles and skin. Furthermore, clinical tests have shown that electroporation can be used to safely deliver vaccines and anticancer gene therapy. Therefore, electroporation delivery of IL-30 gene therapy has great potential to be translated into the clinic. The proposed investigation of the biological mechanism of IL-30's action will lay the foundation for combining IL-30 gene therapy with other approaches to successfully treat liver disease in the future. Our central hypothesis is that low-electric-field electroporation delivery of IL-30 gene therapy reduces and repairs liver injury without causing liver toxicity and that this liver injury reduction and repair is through IL- 30's activation of the NKG2D ligand-NKG2D receptor-mediated death of fibrosis progenitor cells. To test our hypothesis, we will pursue the following two specific aims. Aim 1: Utilize skin-based, low electric field-mediated electroporation delivery of IL-30 gene therapy for treating chronic liver injury. Aim 2: Determine the biological mechanisms underlying the action of IL-30 gene therapy in reducing liver injury. The successful completion of these aims will identify a simple, practical potentially effective method of gene therapy delivery; enable us to translate IL-30 gene therapy to clinical testing in the future; and reveal a novel mechanism by which IL-30 inhibits liver injur. This novel treatment may complement standard therapies to repair liver injury currently believed to be incurable.