关键词: 星形胶质细胞, CRISPR-Cas9, 细胞外囊泡, 纤维蛋白原, 人诱导多能干细胞来源的神经干/前体细胞, ID3, 微小RNA, 神经再生, 神经突生长, 脊髓, 神经再生

Abstract:

Human induced pluripotent stem cell–derived neural stem/progenitor cells are used in cell-replacement and regenerative therapeutic strategies after traumatic central nervous system injury. Traumatic injury alters the host microenvironment, which in turn affects the functionality of transplanted human neural stem/progenitor cells and potentially limits their benefits for neurorepair. However, the underlying mechanisms through which the host environment alters the fate and functionality of transplanted human neural stem/progenitor cells remain poorly understood. Here, we showed that massive deposition of blood-derived fibrinogen in a mouse model of spinal cord injury contributed to an altered lesion environment. Fibrinogen promoted human neural stem/progenitor cell differentiation into reactive astrocytes by activating the BMP receptor signaling pathway and inducing of the transcriptional regulator inhibitor of DNA binding 3. ID3-depleted human neural stem/progenitor cells, generated by CRISPR/Cas9-mediated genome editing, reduced astrocyte formation in response to astrogenic stimuli. Instead, ID3-depleted human neural stem/progenitor cells had a bipolar, immature glial progenitor cell phenotype. These modified cells secreted extracellular vesicles with a distinct miRNA profile that enhanced neurite outgrowth. We conclude that targeting inhibitor of DNA binding 3 in human neural stem/progenitor cells can beneficially modulate their functionality and cell fate in the injured central nervous system toward glial progenitor cells, potentially enhancing their capacity to promote central nervous system repair.

Key words: astrocyte, CRISPR-Cas9, extracellular vesicles, fibrinogen, human iPSC-derived neural stem/progenitor cell, inhibitor of DNA binding 3 (ID3), microRNA, nerve regeneration, neurite outgrowth, spinal cord