Evaluation of modified Interferon alpha mRNA constructs for the treatment of non-melanoma skin cancer.
Evaluation of modified Interferon alpha mRNA constructs for the treatment of non-melanoma skin cancer.
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DOI：
10.1038/s41598-018-31061-w
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发表时间：
2018-08-28
影响因子：
4.6
通讯作者：
Strunk D
中科院分区：
文献类型：
--
作者：
Hochmann S;Mittermeir M;Santic R;Koszik F;Griessner L;Sonderegger AS;Hoffmann T;Russe E;Scheiblhofer S;Weiss R;Mandler M;Schneeberger A;Strunk D
应用体外转录(IVT)信使核糖核酸(MRNA)是一种越来越受欢迎的策略，可以在选择的组织或器官中瞬时产生蛋白质作为治疗药物。在这里，我们将重点放在皮肤上，目的是测试整个人皮肤组织移植技术是否可以用于评估不同的IVT干扰素α(干扰素-α)基因在生物传递后的原位表达效果。根据组织学分析和TUNEL染色，皮肤外植体至少存活5天且完好无损。利用GFP报告基因的表达形式，我们发现在生物递送后主要在表皮表达。在5个序列优化的干扰素-α变异体中，有2个与天然干扰素-α信使核糖核酸转染组相比，显著提高了人皮肤中干扰素-α蛋白的表达。对周围培养上清液中干扰素-α的分泌分析证实了上述结果。我们提供了一种概念证明，将干扰素-mRNAm RNA导入完整的人全层皮肤外植体可以用于体外测试α序列的修改。这种方法可以用来开发基于信使核糖核酸的新型治疗方法，用于治疗常见的表皮皮肤疾病，包括非黑色素瘤皮肤癌，此前干扰素-α蛋白治疗已显示出很强的治疗效果。
Application of in vitro transcribed (IVT) messenger ribonucleic acid (mRNA) is an increasingly popular strategy to transiently produce proteins as therapeutics in a tissue or organ of choice. Here, we focused on the skin and aimed to test if whole human skin tissue explant technology can be used to evaluate the expression efficacy of different IVT Interferon alpha (IFN-α) mRNA constructs in situ, after biolistic delivery. Skin explants were viable and intact for at least five days based on histologic analysis and TUNEL staining. Using GFP reporter mRNA formulations, we found mostly epidermal expression after biolistic delivery. Two out of five sequence-optimized IFN-α mRNA variants resulted in significantly improved IFN-α protein expression in human skin compared to native IFN-α mRNA transfection. IFN-α secretion analysis of the surrounding culture media confirmed these results. We provide a proof-of-concept that IFN-α mRNA delivery into intact human full thickness skin explants can be utilized to test mRNA sequence modifications ex vivo. This approach could be used to develop novel mRNA-based treatments of common epidermal skin conditions including non-melanoma skin cancer, where IFN-α protein therapy has previously shown a strong therapeutic effect.
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影响因子：
12.4
作者：
Kariko, Katalin;Muramatsu, Hiromi;Welsh, Frank A.;Ludwig, Janos;Kato, Hiroki;Akira, Shizuo;Weissman, Drew
通讯作者：
Weissman, Drew
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Sung, Hsing-Wen
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作者：
Kowalska, Joanna;Lewdorowicz, Magdalena;Jemielity, Jacek
通讯作者：
Jemielity, Jacek
影响因子：
4.5
作者：
Grudzien-Nogalska, Ewa;Jemielity, Jacek;Rhoads, Robert E.
通讯作者：
Rhoads, Robert E.