All-trans retinoic acid induces reprogramming of canine dedifferentiated cells into neuron-like cells
All-trans retinoic acid induces reprogramming of canine dedifferentiated cells into neuron-like cells
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DOI：
10.1371/journal.pone.0229892
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发表时间：
2020-03-31
期刊：
影响因子：
3.7
通讯作者：
Sugiya, Hiroshi
中科院分区：
文献类型：
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作者：
Nakano, Rei;Kitanaka, Taku;Sugiya, Hiroshi
细胞身份的规范取决于细胞对生物活性配体序列的暴露。全反式维甲酸（ATRA）影响早期神经元发育，参与神经元谱系重编程。我们之前建立了一种来源于高度同质性成熟脂肪细胞的成纤维样去分化脂肪细胞（DFATs），这种细胞更适合于细胞重编程的研究。犬认知功能障碍与人类认知功能障碍相似，提示犬可能是人类神经元疾病的病理和药理学模型。然而，ATRA对狗神经元重编程的影响仍不清楚。因此，在本研究中，我们研究了ATRA对犬DFAT神经元重编程的影响。ATRA诱导神经元标志物mRNA/蛋白的表达。神经元样细胞表现出Ca 2+内流，伴有去极化（50 mM KCl; 84.75 +/- 4.05%）和Na+通道激活（50 μ M藜芦碱; 96.02 +/- 2.02%）。突触前末梢活动的光学成像和神经递质释放的检测表明，神经元样细胞具有GABA能神经元的特性。全基因组RNA测序分析表明，犬DFAT的转录组谱被有效地重编程为皮质中间神经元谱系。总的来说，ATRA可以从犬DFAT产生功能性GABA能皮质中间神经元样细胞，以> 80%的效率表现出神经元功能。我们进一步证明了JNK 3对犬DFAT中ATRA诱导的神经元重编程的贡献。总之，来自犬DFAT的神经元样细胞可能是细胞移植治疗、体外疾病建模和神经元疾病药物筛选的转化研究的有力工具。
The specification of cell identity depends on the exposure of cells to sequences of bioactive ligands. All-trans retinoic acid (ATRA) affects neuronal development in the early stage, and it is involved in neuronal lineage reprogramming. We previously established a fibroblast-like dedifferentiated fat cells (DFATs) derived from highly homogeneous mature adipocytes, which are more suitable for the study of cellular reprogramming. Canine cognitive dysfunction is similar to human cognitive dysfunction, suggesting that dogs could be a pathological and pharmacological model for human neuronal diseases. However, the effect of ATRA on neuronal reprogramming in dogs has remained unclear. Therefore, in this study, we investigated the effect of ATRA on the neuronal reprogramming of canine DFATs. ATRA induced the expression of neuronal marker mRNA/protein. The neuron-like cells showed Ca2+ influx with depolarization (50 mM KCl; 84.75 +/- 4.05%) and Na+ channel activation (50 mu M veratridine; 96.02 +/- 2.02%). Optical imaging of presynaptic terminal activity and detection of neurotransmitter release showed that the neuron-like cells exhibited the GABAergic neuronal property. Genome-wide RNA-sequencing analysis shows that the transcriptome profile of canine DFATs is effectively reprogrammed towards that of cortical interneuron lineage. Collectively, ATRA can produce functional GABAergic cortical interneuron-like cells from canine DFATs, exhibiting neuronal function with > 80% efficiency. We further demonstrated the contribution of JNK3 to ATRA-induced neuronal reprogramming in canine DFATs. In conclusion, the neuron-like cells from canine DFATs could be a powerful tool for translational research in cell transplantation therapy, in vitro disease modeling, and drug screening for neuronal diseases.