Using siRNA-based spherical nucleic acid nanoparticle conjugates for gene regulation in psoriasis
Using siRNA-based spherical nucleic acid nanoparticle conjugates for gene regulation in psoriasis
复制标题
DOI：
10.1016/j.jconrel.2017.10.034
复制
发表时间：
2017-12-28
影响因子：
10.8
通讯作者：
Tavoosidana, Gholamreza
中科院分区：
文献类型：
--
作者：
Nemati, Houshang;Ghahramani, Mohammad-Hosein;Tavoosidana, Gholamreza
银屑病是一种T细胞介导的具有自身免疫性质的皮肤病，一般在动物中观察不到，缺乏相关的银屑病实验动物模型阻碍了对该病发病机制的研究。小干扰RNA的应用和全身给药为皮肤基因调控提供了许多有效的治疗优势，本研究建立了银屑病IMQ动物模型，并设计了一种安全的融合肽载体--球形核酸金纳米颗粒偶联物，以提高siRNA对细胞和皮肤的穿透能力和对基因调控的靶向性。我们评估了银屑病模型和EGFR siRNA治疗（作为球形核酸纳米颗粒）、表型（红斑、鳞屑、炎症和增厚的体征）、细胞增殖的显微镜评价和CD 3、CD 4和CD 8标志物的免疫化学评价。此外，我们监测了SNA-NC处理A431细胞后EGF和EGFR基因的抑制。在人皮肤细胞中进行qRT-PCR检测，结果表明SNA-NC稳定、无毒。体外实验表明，EGF和EGFR siRNA与球形核酸金纳米颗粒缀合可以显著降低细胞中的基因表达。体内实验表明，通过皮肤局部应用由SNA-NC递送的siRNA可以显著抑制细胞增殖。小鼠背部皮肤的显微镜评价和免疫组化过程证实了SNA-NCs siRNA在银屑病小鼠模型中的抑制作用。这些结果表明，局部应用SNA-NCs siRNA可以通过抑制基因表达和T细胞产生的功能活性来改善银屑病样皮肤病变。
Psoriasis is a T-cell-mediated skin disease with autoimmune nature that is generally not observed in animals, this lack of a relevant experimental animal model of psoriasis has hindered the investigation of pathogenesis of disease. Application and systemic delivery of small interfering RNAs offer many effective therapeutic advantages for gene regulation in the skin.In this study, we present an IMQ animal model of psoriasis and designed a safe fusion peptide carrier, spherical nucleic acid gold nanoparticles conjugate, to improve penetration of the siRNA into the cells and skin and their targeting ability to gene regulation. We evaluated the model of psoriasis and EGFR siRNA treatment (as spherical nucleic acid nanoparticles), phenotypically (signs of erythema, scaling, inflammation and thickening), microscopic evaluation of cell proliferation and immunohistochemically evaluation of CD3, CD4, and CD8 markers. Also, we monitored suppression of EGF & EGFR genes after treatment of A431 cells by SNA-NCs. The expression of genes was validated by qRT-PCR in human skin cells.The results showed that the SNA-NCs were stable and non-toxic. In vitro experiments indicated that EGF & EGFR siRNAs conjugated with spherical nucleic acid gold nanoparticles can significantly reduce gene expression in cells. In vivo experiments showed that the topical application of siRNAs delivered by SNA-NCs through the skin can significantly inhibit the proliferation of cells. Microscopic evaluation of mice back skin and immunohistochemistry process approved Inhibitory effect of SNA-NCs siRNA in the mouse model of psoriasis.Since the proliferation of T cells was crucial for the development of a psoriatic phenotype. These results demonstrate that topical application of SNA-NCs siRNA may improve psoriatic-like skin lesions by suppressing gene expression and functional activity of T cell production.