Development of Synthetic Modulator Enabling Long-Term Propagation and Neurogenesis of Human-Derived Neural Progenitor Cells

Abstract

Neural progenitor cells (NPCs) are important cells for in vitro drug screening and the cell-based therapy for brain-related disorders, which requires well-defined and reproducible culture systems. Current strategy the use of protein growth factors presents challenges in terms of reproducibility and cost. In this study, we have developed a novel DNA-based modulator to regulate FGFR signaling of NPCs, enabling maintenance of the stemness over 50 passages and neurogenesis towards neurons. The DNA-based FGFR-agonist effectively promoted FGFR1 phosphorylation and activated the downstream ERK signaling pathway in FGFR1-positive cells. Using human embryonic stem cell lines, we differentiated them into NPCs and replaced basic fibroblast growth factor (bFGF) in the regulator culture medium with DNA-based FGFR-agonist for artificially elicited FGFR signaling. The results demonstrated that the FGFR-agonist could promote NPCs proliferation and neurosphere formation, recapitulating the function of bFGF. Notably, transcriptomic analysis revealed that FGFR-agonist could customize the stemness-associated transcription program, while decouples the neuronal differentiation program, highly resembling that the native ligand, bFGF. Moreover, our culture condition facilitated the successful propagation of NPCs for over 50 passages, while retaining their ability to efficiently differentiate into neurons. Overall, our approach provides a highly effective method for expanding NPCs, offering new opportunities for disease-in-dish research and drug screening for neural degeneration.