Direct conversion of mouse fibroblasts to self-renewing, tripotent neural precursor cells
Direct conversion of mouse fibroblasts to self-renewing, tripotent neural precursor cells
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DOI：
10.1073/pnas.1121003109
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发表时间：
2012-02-14
影响因子：
11.1
通讯作者：
Wernig, Marius
中科院分区：
文献类型：
--
作者：
Lujan, Ernesto;Chanda, Soham;Wernig, Marius
我们最近表明，定义的转录因子集足以将小鼠和人成纤维细胞直接转化为类似于功能性神经元的细胞，称为“诱导神经元”（iN）细胞。然而，对于某些应用，期望将成纤维细胞转化为增殖性神经前体细胞（NPC）而不是神经元。我们假设NPC样细胞可以使用用于产生iN细胞的相同的主要方法来诱导。为了实现这一目标，我们用一组在NPC中高度表达的11个转录因子感染来自Sox 2-EGFP小鼠的小鼠胚胎成纤维细胞。转基因诱导后24天，Sox 2-EGFP（+）集落出现，表达NPC特异性基因，并分化为神经元和星形胶质细胞。使用逐步消除，我们发现Sox 2和FoxG 1能够产生克隆自我更新，双能诱导的NPC，产生星形胶质细胞和功能性神经元。当我们将含有Pou和同源框结构域的转录因子Brn 2加入Sox 2和FoxG 1时，我们能够诱导三能NPC，这些NPC不仅可以分化为神经元和星形胶质细胞，还可以分化为少突胶质细胞。转录因子FoxG 1和Brn 2单独也能够诱导NPC样细胞;然而，这些细胞产生不太成熟的神经元，尽管它们确实产生星形胶质细胞，甚至少突胶质细胞，能够整合到髓鞘发育不良的Shiverer脑中。我们的数据表明，使用靶细胞类型特异性转录因子的直接谱系重编程可用于诱导NPC样细胞，这些细胞可能用于脑或脊髓中基于自体细胞移植的治疗。
We recently showed that defined sets of transcription factors are sufficient to convert mouse and human fibroblasts directly into cells resembling functional neurons, referred to as "induced neuronal" (iN) cells. For some applications however, it would be desirable to convert fibroblasts into proliferative neural precursor cells (NPCs) instead of neurons. We hypothesized that NPC-like cells may be induced using the same principal approach used for generating iN cells. Toward this goal, we infected mouse embryonic fibroblasts derived from Sox2-EGFP mice with a set of 11 transcription factors highly expressed in NPCs. Twenty-four days after transgene induction, Sox2-EGFP(+) colonies emerged that expressed NPC-specific genes and differentiated into neuronal and astrocytic cells. Using stepwise elimination, we found that Sox2 and FoxG1 are capable of generating clonal self-renewing, bipotent induced NPCs that gave rise to astrocytes and functional neurons. When we added the Pou and Homeobox domain-containing transcription factor Brn2 to Sox2 and FoxG1, we were able to induce tripotent NPCs that could be differentiated not only into neurons and astrocytes but also into oligodendrocytes. The transcription factors FoxG1 and Brn2 alone also were capable of inducing NPC-like cells; however, these cells generated less mature neurons, although they did produce astrocytes and even oligodendrocytes capable of integration into dysmyelinated Shiverer brain. Our data demonstrate that direct lineage reprogramming using target cell-type-specific transcription factors can be used to induce NPC-like cells that potentially could be used for autologous cell transplantation-based therapies in the brain or spinal cord.