Pan Zhimin, Oh Jinsoo, Huang Lu, Zeng Zhaoxun, Duan Pingguo, Li Zhiyun, Yun Yeomin, Kim Janghwan, Ha Yoon, Cao Kai
Spine Center, The Second Affiliated Hospital of Nanchang University, Nanchang, China.
Department of Neurosurgery, Spine and Spinal Cord Institute, Yonsei University College of Medicine, Seoul, Republic of Korea.
Ann Transl Med. 2020 Aug;8(15):933. doi: 10.21037/atm-20-3871.
Spinal cord injury (SCI) tends to damage neural tissue and generate a hypoxic environment. Studies have confirmed that single therapy with gene or stem cells is inefficient, but research into combining stem cells and gene therapy in treating tissue damage has been undertaken to overcome the related limitations, which include low gene delivery efficiency and therapeutic outcome. Thus, a combination of stem cells, gene therapy, and a hypoxia-specific system may be useful for the reconstruction of SCI.
To synergistically treat SCI, a combined platform using a hypoxia/neuron-inducible gene expression system (HNIS) and human induced-neural stem cells (hiNSCs) produced by direct reprogramming was designed. Sox2- or nestin-positive hiNSCs were differentiated to Tuj1-, MAP2-, or NeuN-positive neurons.
HNIS showed consistent hypoxia/neuron-specific gene expression in hiNSCs cultured under hypoxia. In particular, the HNIS-hiNSC combined platform revealed a complex pattern with higher gene expression compared with a single platform. In addition, we found that an optimal combination of small molecules, such as CHIR99021, valproic acid (VPA), glycogen synthase kinase-3β (GSK3β), and histone deacetylase (HDAC) inhibitors, could significantly enhance gene expression with HNIS-hiNSCs in the hypoxic environment.
This experiment demonstrated that HNIS-hiNSCs combined with GSK3 and HDAC inhibitors may present another promising strategy in the treatment of SCI.
脊髓损伤（SCI）往往会损害神经组织并产生缺氧环境。研究证实，单一的基因或干细胞治疗效率低下，但已开展将干细胞与基因治疗相结合以治疗组织损伤的研究，以克服包括低基因递送效率和治疗效果在内的相关局限性。因此，干细胞、基因治疗和缺氧特异性系统的组合可能有助于脊髓损伤的重建。
为了协同治疗脊髓损伤，设计了一种使用缺氧/神经元诱导基因表达系统（HNIS）和通过直接重编程产生的人诱导神经干细胞（hiNSCs）的联合平台。将Sox2或巢蛋白阳性的hiNSCs分化为Tuj1、MAP2或NeuN阳性神经元。
HNIS在缺氧条件下培养的hiNSCs中显示出一致的缺氧/神经元特异性基因表达。特别是，与单一平台相比，HNIS-hiNSC联合平台显示出更复杂的模式，基因表达更高。此外，我们发现小分子的最佳组合，如CHIR99021、丙戊酸（VPA）、糖原合酶激酶-3β（GSK3β）和组蛋白去乙酰化酶（HDAC）抑制剂，可以在缺氧环境中显著增强HNIS-hiNSCs的基因表达。
本实验表明，HNIS-hiNSCs与GSK3和HDAC抑制剂联合使用可能是治疗脊髓损伤的另一种有前景的策略。
