Towards the development of a RNAi-based topical treatment for psoriasis: Proof-of-concept in a 3D psoriasis skin model
Towards the development of a RNAi-based topical treatment for psoriasis: Proof-of-concept in a 3D psoriasis skin model
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DOI：
10.1111/exd.13414
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发表时间：
2018-05-01
影响因子：
3.6
通讯作者：
Lambert, Jo
中科院分区：
文献类型：
--
作者：
Desmet, Eline;Van Gele, Mireille;Lambert, Jo
RNA干扰已经成为治疗基因沉默的有力工具，因为它提供了沉默几乎任何已知的致病基因的可能性。然而，RNAi分子的体内递送受到其不利的物理化学特性和对内源性酶降解的敏感性的阻碍。为了克服这些局限性，我们最近开发了一种弹性脂质体制剂，称为DDC 642，作为皮肤疾病治疗性RNAi分子的局部递送系统。在这项研究中，我们使用3种不同的体外模型验证了DDC 642包封的RNAi分子在治疗银屑病中的疗效：标准化角质形成细胞单层培养物、银屑病诱导的角质形成细胞和银屑病重建的皮肤模型。选择了已知在银屑病病变中上调的四种基因（IL 22 RA 1、KRT 17、DEFB 4和TSLP），从而选择了银屑病发病机制中的关键参与者。此外，探索了在银屑病的局部治疗中使用组合siRNA疗法的可能性。结果表明，在mRNA和蛋白质水平上，每个不同的靶标都成功地基因沉默。此外，siRNA-DDC 642治疗导致特定银屑病标志物的表达减少，表明其在未来治疗方法中的潜力。所检查的siRNA组合（即KRT 17、DEFB 4和TSLP的同时敲低）显示TSLP表达的降低增强，而K17蛋白表达的降低在银屑病角质形成细胞中受损。尽管本文研究的siRNA组合仍可以进一步改进，但我们的研究已经在体外证明了同时靶向多个基因的临床潜力，每个基因在复杂疾病如银屑病中发挥不同的作用。
RNA interference has emerged as a powerful tool for therapeutic gene silencing, as it offers the possibility to silence virtually any known pathology-causing gene. However, in vivo delivery of RNAi molecules is hampered by their unfavourable physicochemical characteristics and susceptibility to degradation by endogenous enzymes. To overcome these limitations, we recently developed an elastic liposomal formulation, called DDC642, as topical delivery system of therapeutic RNAi molecules for skin disorders. In this study, we validated the therapeutic efficacy of DDC642-encapsulated RNAi molecules in the treatment of psoriasis using 3 different in vitro models: a standardized keratinocyte monolayer culture, psoriasis-induced keratinocytes and a psoriasis-reconstructed skin model. Four genes (IL22RA1, KRT17, DEFB4 and TSLP), known to be upregulated in psoriatic lesions, and thereby key players in psoriasis pathogenesis were selected. Moreover, the possibility of using a combined siRNA therapy in the topical treatment of psoriasis was explored. Results indicate a successful gene silencing of each different target, both at mRNA and protein levels. Additionally, siRNA-DDC642 treatment resulted in a reduced expression of specific psoriasis markers, indicating their potential in future therapeutic approach. The examined siRNA combination (ie simultaneous knockdown of KRT17, DEFB4 and TSLP) showed an enhanced reduction in TSLP expression, whereas the decrease in K17 protein expression was impaired in psoriatic keratinocytes. Although the here examined siRNA combination could still be further improved, our study proved already in vitro the clinical potential of targeting multiple genes at once, each playing a different role in a complex disease such as psoriasis.