High-throughput screening of clinically approved drugs that prime nonviral gene delivery to human Mesenchymal stem cells
High-throughput screening of clinically approved drugs that prime nonviral gene delivery to human Mesenchymal stem cells
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DOI：
10.1186/s13036-020-00238-1
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发表时间：
2020-05-19
影响因子：
5.6
通讯作者：
Pannier, Angela K.
中科院分区：
文献类型：
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作者：
Kozisek, Tyler;Hamann, Andrew;Pannier, Angela K.
背景人骨髓间充质干细胞（humanmesenchymalstemcells，hMSCs）因其独特的生物学特性在细胞治疗学中的应用受到广泛关注，然而，通过基因修饰可以增强hMSCs的内在治疗特性。病毒转导是有效的，但存在安全性问题。相反，非病毒基因递送虽然比病毒更安全，但效率低且具有细胞毒性，特别是在hMSC中。为了解决非病毒基因递送到hMSCs的缺点，我们的实验室先前已经证明，用糖皮质激素地塞米松对hMSCs进行药理学“引发”可以通过调节转染诱导的细胞毒性来显著增加hMSCs中的转染。这项工作旨在通过筛选707种FDA批准的药物来建立hMSCs的转染引发化合物库，这些药物属于不同的药物类别，来自NIH临床收藏的四种浓度，用于调节来自两个人供体的脂肪来源的hMSCs的非病毒基因递送的能力。结果分析用荧光转基因转染的细胞的显微镜图像，以鉴定显著影响hMSC转染而没有显著毒性的化合物。增加两种供体间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，糖皮质激素，增加转基因生产18倍以上未引发转染。此外，降低两种供体之间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-与未引发的转染相比为41倍。结论我们的NCC筛选是第一个高通量和药物再利用的方法，以确定非病毒基因递送引发化合物在两个供体的hMSCs。引发化合物和类确定在这个屏幕上表明，增殖，线粒体功能和细胞凋亡的调制是至关重要的增强非病毒基因传递到hMSCs。
Background 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. Results 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. Conclusions 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.