Human mesenchymal stem cells (hMSCs) are intensely researched for applications in cell therapeutics due to their unique properties, however, intrinsic therapeutic properties of hMSCs could be enhanced by genetic modification. Viral transduction is efficient, but suffers from safety issues. Conversely, nonviral gene delivery, while safer compared to viral, suffers from inefficiency and cytotoxicity, especially in hMSCs. To address the shortcomings of nonviral gene delivery to hMSCs, our lab has previously demonstrated that pharmacological ‘priming’ of hMSCs with the glucocorticoid dexamethasone can significantly increase transfection in hMSCs by modulating transfection-induced cytotoxicity. This work seeks to establish a library of transfection priming compounds for hMSCs by screening 707 FDA-approved drugs, belonging to diverse drug classes, from the NIH Clinical Collection at four concentrations for their ability to modulate nonviral gene delivery to adipose-derived hMSCs from two human donors. Microscope images of cells transfected with a fluorescent transgene were analyzed in order to identify compounds that significantly affected hMSC transfection without significant toxicity. Compound classes that increased transfection across both donors included glucocorticoids, antibiotics, and antihypertensives. Notably, clobetasol propionate, a glucocorticoid, increased transgene production 18-fold over unprimed transfection. Furthermore, compound classes that decreased transfection across both donors included flavonoids, antibiotics, and antihypertensives, with the flavonoid epigallocatechin gallate decreasing transgene production − 41-fold compared to unprimed transfection. Our screen of the NCC is the first high-throughput and drug-repurposing approach to identify nonviral gene delivery priming compounds in two donors of hMSCs. Priming compounds and classes identified in this screen suggest that modulation of proliferation, mitochondrial function, and apoptosis is vital for enhancing nonviral gene delivery to hMSCs.

中文翻译：

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对临床批准的药物进行高通量筛选，这些药物可将非病毒基因递送至人间充质干细胞。

人类间充质干细胞 (hMSCs) 因其独特的特性而被深入研究用于细胞治疗，然而，hMSCs 的内在治疗特性可以通过基因修饰来增强。病毒转导是有效的，但存在安全问题。相反，与病毒相比，非病毒基因传递虽然更安全，但存在效率低下和细胞毒性，尤其是在 hMSCs 中。为了解决非病毒基因向 hMSCs 传递的缺点，我们的实验室之前已经证明，用糖皮质激素地塞米松对 hMSCs 进行药理学“启动”可以通过调节转染诱导的细胞毒性显着增加 hMSCs 的转染。这项工作旨在通过筛选属于不同药物类别的 707 种 FDA 批准的药物，为 hMSCs 建立转染引发化合物库，来自美国国立卫生研究院临床收藏的四种浓度，因为它们能够调节来自两个人类供体的非病毒基因向脂肪来源的 hMSCs 的传递。分析了用荧光转基因转染的细胞的显微镜图像，以确定显着影响 hMSC 转染而没有显着毒性的化合物。增加两个供体转染的化合物类别包括糖皮质激素、抗生素和抗高血压药。值得注意的是，丙酸氯倍他索（一种糖皮质激素）比未引物转染增加了 18 倍的转基因产量。此外，减少两个供体转染的化合物类别包括类黄酮、抗生素和抗高血压药，其中类黄酮表没食子儿茶素没食子酸酯降低转基因产量 - 与未引物转染相比减少了 41 倍。我们的 NCC 筛选是第一个高通量和药物再利用方法，用于识别两个 hMSCs 供体中的非病毒基因递送引发化合物。此筛选中确定的引发化合物和类别表明，增殖、线粒体功能和细胞凋亡的调节对于增强非病毒基因向 hMSCs 的传递至关重要。