Hnf1β、Emx2、Pax8和Hnf4的表达水平和配比影响诱导肾小管上皮细胞的直接重编程。

IF 4 Q2 CELL & TISSUE ENGINEERING
Xueli Hu, Jianjian Sun, Meng Wan, Bianhong Zhang, Linhui Wang, Tao P Zhong
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引用次数: 0

摘要

从成纤维细胞生成诱导肾上皮细胞(iRECs)为肾脏疾病建模和肾脏再生提供了巨大的机遇。然而,目前生成 iRECs 的重编程效率较低,阻碍了其潜在的治疗应用和再生方法。部分原因可能是转导成纤维细胞中的重编程因子(RFs)Hnf1β(H1)、Emx2(E)、Pax8(P)和Hnf4α(H4)表达不均且不平衡。在这里,我们建立了一种先进的逆转录病毒载体系统,它能高水平地表达 H1、E、P 和 H4,而且这些转录本与由 P2A 分离出来的双组分转录本的比例截然不同。利用携带Cdh16-Cre; mT/mG等位基因的小鼠胚胎成纤维细胞(MEF),通过直接监测单细胞命运转换来进行iREC重编程。为了诱导iREC重编程,产生了三组双核RF组合,包括H1E/H4P、H1H4/EP和H1P/H4E。每种RF组合都能产生不同的H1、E、P和H4表达水平以及不同的重编程效率。与单独的 H1、E、P 和 H4 慢病毒转导相比,理想的 H1E/H4P 组合能以平衡的化学计量产生高表达水平的 RF,从而大大提高 iRECs 的效率和质量。我们发现,H1E/H4P 诱导的 iRECs 表现出肾小管上皮细胞的优异特征,如表达肾小管特异性基因、具有内吞傲慢活性以及沿着脱细胞肾支架组装成肾小管。这项研究确立了 H1E/H4P 盒作为未来 iREC 研究和再生医学的宝贵平台的地位。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Expression levels and stoichiometry of Hnf1β, Emx2, Pax8 and Hnf4 influence direct reprogramming of induced renal tubular epithelial cells.

Generation of induced renal epithelial cells (iRECs) from fibroblasts offers great opportunities for renal disease modeling and kidney regeneration. However, the low reprogramming efficiency of the current approach to generate iRECs has hindered potential therapeutic application and regenerative approach. This could be in part attributed to heterogeneous and unbalanced expression of reprogramming factors (RFs) Hnf1β (H1), Emx2 (E), Pax8 (P), and Hnf4α (H4) in transduced fibroblasts. Here, we establish an advanced retroviral vector system that expresses H1, E, P, and H4 in high levels and distinct ratios from bicistronic transcripts separated by P2A. Mouse embryonic fibroblasts (MEFs) harboring Cdh16-Cre; mT/mG allele are utilized to conduct iREC reprogramming via directly monitoring single cell fate conversion. Three sets of bicistronic RF combinations including H1E/H4P, H1H4/EP, and H1P/H4E have been generated to induce iREC reprogramming. Each of the RF combinations gives rise to distinct H1, E, P, and H4 expression levels and different reprogramming efficiencies. The desired H1E/H4P combination that results in high expression levels of RFs with balanced stoichiometry. substantially enhances the efficiency and quality of iRECs compared with transduction of separate H1, E, P, and H4 lentiviruses. We find that H1E/H4P-induced iRECs exhibit the superior features of renal tubular epithelial cells, as evidenced by expressing renal tubular-specific genes, possessing endocytotic arrogation activity and assembling into tubules along decellularized kidney scaffolds. This study establishes H1E/H4P cassette as a valuable platform for future iREC studies and regenerative medicine.

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来源期刊
Cell Regeneration
Cell Regeneration Biochemistry, Genetics and Molecular Biology-Cell Biology
CiteScore
5.80
自引率
0.00%
发文量
42
审稿时长
35 days
期刊介绍: Cell Regeneration aims to provide a worldwide platform for researches on stem cells and regenerative biology to develop basic science and to foster its clinical translation in medicine. Cell Regeneration welcomes reports on novel discoveries, theories, methods, technologies, and products in the field of stem cells and regenerative research, the journal is interested, but not limited to the following topics: ◎ Embryonic stem cells ◎ Induced pluripotent stem cells ◎ Tissue-specific stem cells ◎ Tissue or organ regeneration ◎ Methodology ◎ Biomaterials and regeneration ◎ Clinical translation or application in medicine
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