As the human lifespan has increased due to developments in medical technology, the number of patients with neurological diseases has rapidly increased. Therefore, studies on effective treatments for neurological diseases are becoming increasingly important. To perform these studies, it is essential to obtain a large number of patient-derived neural cells. The purpose of the present study was to establish a technology that allows the high-efficiency generation of genetically stable, direct-conversion-derived neural stem cells (dcNSCs) through the expression of a new combination of reprogramming factors, including a proto-oncogene. Specifically, human c-MYC proto-oncogene and the human SOX2 gene were overexpressed in a precisely controlled manner in various human somatic cells. As a result, the direct conversion into multipotent dcNSCs occurred only when the cells were treated with an MOI of 1 of hc-MYC proto-oncogene and hSOX2 retrovirus. When MOIs of 5 or 10 were utilized, distinct results were obtained. In addition, the pluripotency was bypassed during this process. Notably, as the MOI used to treat the cells increased, expression of the p53 tumor suppressor gene, which is typically a reprogramming hurdle, increased proportionately. Interestingly, p53 was genetically stable in dcNSCs generated through direct conversion into a low p53 expression state. In the present study, generation of genetically stable dcNSCs using direct conversion was optimized by precisely controlling the overexpression of a proto-oncogene. This method could be utilized in future studies, such as in vitro drug screening using generated dcNSCs. In addition, this method could be effectively utilized in studies on direct conversion into other types of target cells.

随着人类寿命随着医疗技术的发展而增加，患有神经系统疾病的患者数量迅速增加。因此，对神经疾病的有效治疗的研究变得越来越重要。为了进行这些研究，获得大量患者来源的神经细胞是必不可少的。本研究的目的是建立一种技术，该技术可通过表达包括原癌基因在内的重编程因子的新组合来高效生成遗传稳定的直接转化衍生的神经干细胞（dcNSC）。具体而言，人类c-MYC原癌基因和人类SOX2该基因在各种人体细胞中以精确控制的方式过表达。结果，仅当用hc-MYC原癌基因和hSOX2逆转录病毒的MOI为1处理细胞时，才直接转化为多能dcNSC 。当使用5或10的MOI时，获得了不同的结果。另外，在该过程中多能性被绕过。值得注意的是，随着用于治疗细胞的MOI的增加，通常是重编程障碍的p53抑癌基因的表达也成比例地增加。有趣的是，p53在通过直接转化为低p53表达状态而产生的dcNSC中具有遗传稳定性。在本研究中，通过直接控制原癌基因的过表达，优化了使用直接转化的遗传稳定的dcNSCs的产生。此方法可用于将来的研究，例如使用生成的dcNSC进行体外药物筛选。此外，该方法可有效地用于直接转化为其他类型靶细胞的研究中。