Direct reprogramming of epidermal cells to neural crest derivatives for cell therapies

摘要 我们实验室最近发现，出生后的人表皮角质形成细胞(KC)可以被重新编程 在没有基因导入转录因子或重新编程的情况下进入神经峰(NC)命运 多能状态。KC派生NC(KC-NC)可以被诱骗以区分为所有功能NC 衍生产品包括外周神经元、黑素细胞、雪旺细胞和间充质干细胞衍生产品 (骨细胞、软骨细胞、脂肪细胞和平滑肌细胞)。在移植到鸡胚中后， KC-NC沿着刻板的路径迁移，并产生了多种NC衍生物。在此，我们建议 将我们的发现扩展到成人表皮角质形成细胞，提供了对NC的机制理解 重新编程过程，并演示了使用KC-NC来源的雪旺细胞治疗 脱髓鞘疾病。在目标1中，我们将确定成人表皮角质形成细胞是否可以 在特定条件下重新编程为神经脊干细胞。目标2将研究KC的机制 重新编程为NC命运。在目标3中，我们将开发一种高效的策略，将KC-NC区分为 雪旺细胞。KC-NC雪旺细胞将用于治疗脱髓鞘疾病 先天性髓鞘减少症的小鼠模型已成为评估 髓鞘细胞制剂。这项工作代表了干细胞生物学的范式转变，正如它展示的那样 人类表皮细胞的可塑性将被重新编程为共同发育来源的细胞-两者 来自外胚层--没有基因改造，在特定的培养条件下。最后，我们的 这项工作有可能提供一种新的来源，丰富的，容易获得的自体干细胞 治疗神经退行性疾病，细胞来源仍然是一个严重的障碍 阻碍了细胞治疗的方法。

ABSTRACT Our laboratory recently discovered that postnatal human epidermal keratinocytes (KC) could be reprogrammed into a neural crest (NC) fate without genetic introduction of transcription factors or reprogramming to the pluripotent state. The KC-derived NC (KC-NC) could be coaxed to differentiate into all functional NC derivatives including peripheral neurons, melanocytes, Schwann cells and mesenchymal stem cell derivatives (osteocytes, chondrocytes, adipocytes and smooth muscle cells). Upon transplantation into chicken embryos, KC-NC migrated along stereotypical pathways and gave rise to multiple NC derivatives. Here we propose to extend our findings to adult epidermal keratinocytes, provide mechanistic understanding of the NC reprograming process, and demonstrate the use of KC-NC derived Schwann cells for treatment of demyelinating disease. In aim 1, we will determine whether human adult epidermal keratinocytes can be reprogrammed into neural crest stem cells under defined conditions. Aim 2 will study the mechanism of KC reprogramming into NC fate. In aim 3, we will develop a highly efficient strategy to differentiate KC-NC into Schwann cells. The KC-NC Schwann cells will be employed for the treatment of demyelinating disease using a mouse model of congenital hypomyelinating disease that has become the gold standard for the assessment of myelinating cell preparations. This work represents a paradigm shift in stem cell biology as it demonstrates the plasticity of human epidermal cells to be reprogrammed into cells of common developmental origin – both originate from the ectoderm - without genetic modification and under defined culture conditions. Finally, our work has the potential to provide a novel source of abundant, readily accessible, autologous stem cells for treatment of neurodegenerative diseases, for which cell sourcing remains a severe impediment hampering cell therapy approaches.