Central Laboratory, School of Medicine, First Affiliated Hospital, Zhejiang University, Hangzhou, China.
Institute of Pharmacology and Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.
CNS Neurosci Ther. 2018 Sep;24(9):801-809. doi: 10.1111/cns.12821. Epub 2018 Feb 17.
The cholinergic deficit is thought to underlie progressed cognitive decline in Alzheimer Disease. The lineage reprogramming of somatic cells into cholinergic neurons may provide strategies toward cell-based therapy of neurodegenerative diseases.
Here, we found that a combination of neuronal transcription factors, including Ascl1, Myt1l, Brn2, Tlx3, and miR124 (5Fs) were capable of directly converting human brain vascular pericytes (HBVPs) into cholinergic neuronal cells. Intriguingly, the inducible effect screening of reprogramming factors showed that a single reprogramming factor, Myt1l, induced cells to exhibit similarly positive staining for Tuj1, MAP2, ChAT, and VAChT upon lentivirus infection with the 5Fs after 30 days. HBVP-converted neurons were rarely labeled even after long-term incubation with BrdU staining, suggesting that induced neurons were directly converted from HBVPs rather than passing through a proliferative state. In addition, the overexpression of Myt1l induced the elevation of Ascl1, Brn2, and Ngn2 levels that contributed to reprogramming.
Our findings provided proof of the principle that cholinergic neurons could be produced from HBVPs by reprogramming factor-mediated fate instruction. Myt1l was a critical mediator of induced neuron cell reprogramming. HBVPs represent another excellent alternative cell resource for cell-based therapy to treat neurodegenerative disease.
胆碱能缺陷被认为是阿尔茨海默病认知能力进行性下降的基础。体细胞向胆碱能神经元的谱系重编程可能为神经退行性疾病的基于细胞的治疗提供策略。
在这里，我们发现包括神经元转录因子 Ascl1、Myt1l、Brn2、Tlx3 和 miR124（5Fs）在内的组合能够直接将人脑血管周细胞（HBVP）转化为胆碱能神经元细胞。有趣的是，重编程因子的诱导效应筛选表明，单个重编程因子 Myt1l 在感染 5Fs 后的第 30 天，通过慢病毒感染，诱导细胞表现出类似的 Tuj1、MAP2、ChAT 和 VAChT 的阳性染色。即使经过长时间的 BrdU 染色孵育，HBVP 转化的神经元也很少被标记，这表明诱导神经元是直接从 HBVP 转化而来，而不是经过增殖状态。此外，Myt1l 的过表达诱导了 Ascl1、Brn2 和 Ngn2 水平的升高，这有助于重编程。
我们的研究结果提供了证据，证明了通过重编程因子介导的命运指令，可以从 HBVP 产生胆碱能神经元。Myt1l 是诱导神经元细胞重编程的关键介质。HBVP 代表了另一种用于神经退行性疾病细胞治疗的优秀替代细胞资源。
