中国神经再生研究(英文版) ›› 2023, Vol. 18 ›› Issue (5): 1090-1098.doi: 10.4103/1673-5374.355820

• 原著:退行性病与再生 • 上一篇    下一篇

小分子化合物可在低氧环境下诱导成纤维细胞重编程为神经祖细胞治疗帕金森病

  

  • 出版日期:2023-05-15 发布日期:2022-11-01
  • 基金资助:
    中国国家自然科学基金项目(81771381);安徽省重点研发项目(2022e07020030,2022e07020032);蚌埠医学院科研项目(2021byfy002);安徽省高等学校自然科学基金项目(KJ2021ZD0085);中国大学生创新计划项目(202110367043,20211036744)

Neural progenitor cells derived from fibroblasts induced by small molecule compounds under hypoxia for treatment of Parkinson’s disease in rats

Yu Guo1, #, Yuan-Yuan Wang1, #, Ting-Ting Sun1, #, Jia-Jia Xu1, Pan Yang1, Cai-Yun Ma1, 2, Wei-Jun Guan2, Chun-Jing Wang1, Gao-Feng Liu1, *, Chang-Qing Liu1, 3, *#br#   

  1. 1School of Laboratory Medicine, School of Life Sciences, Bengbu Medical College, Bengbu, Anhui Province, China;  2National Germplasm Resource Center for Domestic Animals, Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing, China;  3Department of Neuroscience, University of Connecticut Health Center, Farmington, CT, USA
  • Online:2023-05-15 Published:2022-11-01
  • Contact: Chang-Qing Liu, PhD, lcq7813@bbmc.edu.cn; Gao-Feng Liu, MS, lgfmy@bbmc.edu.cn.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 81771381 (to CQL); Anhui Provincial Key Research and Development Project, Nos. 2022e07020030 (to CQL), 2022e07020032 (to YG); Science Research Project of Bengbu Medical College, No. 2021byfy002 (to CQL); the Natural Science Foundation of the Higher Education Institutions of Anhui Province, No. KJ2021ZD0085 (to CJW); and the Undergraduate Innovative Training Program of China, Nos. 202110367043 (to CQL), 202110367044 (to YG).

摘要:

神经祖细胞能够自我更新和分化为神经细胞谱系,其可作为神经退行性疾病的细胞移植治疗的种子细胞,但是内源性神经前体细胞数量较少,不能满足中枢神经系统损伤的需要。既往研究发现,化学混合物 VCR(丙戊酸、CHIR99021 和 Repsox)可在低氧环境中诱导小鼠成纤维细胞分化为神经祖细胞。基于此,实验在5% O2缺氧条件下以VCR (0.5 mM Valproic acid, 3 μM CHIR99021, and 1 μM Repsox)诱导大鼠胚胎成纤维细胞重编程为神经祖细胞。这些神经前体细胞具有典型的神经胚球结构,可表达神经前体细胞标记物Nestin,Sox2和Pax6,同时也可在体外分化为星形胶质细胞和功能性神经元,基因表达谱方面与大鼠脑源性神经干细胞相似。继而将诱导的神经祖细胞立体定向移植到6-羟基多巴胺注射建立的帕金森病大鼠模型的大脑黑质中。可见移植的神经祖细胞在体内长期存活,远距离迁移,并能分化为多种功能性神经元和神经胶质细胞。此外,移植了诱导神经祖细胞的帕金森病大鼠行为缺陷明显改善。表明大鼠成纤维细胞可通过低氧联合VCR诱导直接转化为神经祖细胞,并作为帕金森病移植治疗的有吸引力的供体来源。

https://orcid.org/0000-0002-1059-6528 (Chang-Qing Liu)

关键词: 神经祖细胞, 细胞重编程, 小分子化合物, 缺氧, 帕金森病, 细胞移植, 黑质, 神经功能

Abstract: Neural progenitor cells (NPCs) capable of self-renewal and differentiation into neural cell lineages offer broad prospects for cell therapy for neurodegenerative diseases. However, cell therapy based on NPC transplantation is limited by the inability to acquire sufficient quantities of NPCs. Previous studies have found that a chemical cocktail of valproic acid, CHIR99021, and Repsox (VCR) promotes mouse fibroblasts to differentiate into NPCs under hypoxic conditions. Therefore, we used VCR (0.5 mM valproic acid, 3 μM CHIR99021, and 1 μM Repsox) to induce the reprogramming of rat embryonic fibroblasts into NPCs under a hypoxic condition (5%). These NPCs exhibited typical neurosphere-like structures that can express NPC markers, such as Nestin, SRY-box transcription factor 2, and paired box 6 (Pax6), and could also differentiate into multiple types of functional neurons and astrocytes in vitro. They had similar gene expression profiles to those of rat brain-derived neural stem cells. Subsequently, the chemically-induced NPCs (ciNPCs) were stereotactically transplanted into the substantia nigra of 6-hydroxydopamine-lesioned parkinsonian rats. We found that the ciNPCs exhibited long-term survival, migrated long distances, and differentiated into multiple types of functional neurons and glial cells in vivo. Moreover, the parkinsonian behavioral defects of the parkinsonian model rats grafted with ciNPCs showed remarkable functional recovery. These findings suggest that rat fibroblasts can be directly transformed into NPCs using a chemical cocktail of VCR without introducing exogenous factors, which may be an attractive donor material for transplantation therapy for Parkinson’s disease. 

Key words: cell reprogramming, cell transplantation, hypoxia, neural progenitor cells, neurological function, Parkinson’s disease, small molecule compounds, substantia nigra