The telomerase inhibitor AZT enhances differentiation and prevents overgrowth of human pluripotent stem cell–derived neural progenitors

Abstract

Human pluripotent stem cell (hPSC)-based cell-replacement therapy has emerged as a promising approach for addressing numerous neurological diseases. However, hPSC transplantation has the potential to cause human cell overgrowth and cancer, which represents a major obstacle to implementing hPSC-based therapies. Inhibition of the overgrowth of transplanted cells could help reduce the risk for hPSC transplantation-induced tumorigenesis. In this study, we report that the telomerase inhibitor azidothymidine (3'-azido-3'-deoxythymidine; AZT) enhances the differentiation of cortical neurons and significantly suppresses the proliferation of hPSC-derived cortical progenitors. Using human embryonic stem cells and induced pluripotent stem cells in culture, we found that AZT effectively reduces the number of dividing progenitors without inducing cell death. Furthermore, AZT promoted differentiation of cortical progenitors and maturation of cortical neurons. Of note, AZT-pretreated, hPSC-derived neural progenitors exhibited decreased proliferation and increased differentiation into cortical neurons when transplanted into the mouse brain. In summary, our findings indicate that AZT prevents the overgrowth of hPSC-derived neural precursors and enhances the differentiation of cortical neurons in both cell cultures and hPSC-transplanted mouse brain. We propose that our work could inform clinical applications of hPSC-based cell therapy.

Tracking cell-type-specific temporal dynamics in human and mouse brains

“…The augmented recovery of EdU+ cells from young brains facilitated the identification of rare proliferating cell types previously overlooked, such as arachnoid barrier cells located within the blood- cerebrospinal fluid (CSF) barrier 64 and pituitary stem cells from the pituitary gland 65 ( Figure S5 ). Notably, the telomerase inhibitor resulted in enriched intermediate neuroblasts, associated with reduced fraction of early progenitor cells and late-stage differentiated cells along the neurogenesis trajectory ( Figures S4K and S4L ), consistent with previous studies 54 , 66 , 67 ( Figure S4M ). This analysis demonstrates the potential of TrackerSci to aid in characterizing cell proliferation and differentiation dynamics upon in vivo perturbations.…”

Telomerase-Mediated Self-Assembly of DNA Network in Cancer Cells Enabling Mitochondrial Interference

“…Next, MCF-7 cells were preincubated with TE inhibitor AZT − with different concentrations for 24 h, respectively, and then incubated with TPP-Y and TAMRA-labeled and BHQ2-modified T-L-DNA (denoted as TPP-Y-T-L) for 12 h. The images of CLSM showed that the fluorescence in the MCF-7 cells gradually decreased as the AZT concentration increased (Figure S14). This result revealed that the disassembly of T-L-DNA depended on the TE activity.…”

Electrophysiological Profile of Differentiating Human Central Canal Ependymal Stem-Progenitor Cells

“…To that end, future studies will look into alternative media specifically designed for primary cell cultures. Early literature has found that BrainPhys neuronal media from StemCell Technologies promotes synaptic activity, neuron differentiation, and cell health of primary human stem cells over the long term, with cultures lasting up to 18 weeks 179,180 . Further, the criteria used for cell selection may be biasing for cells at certain developmental stages.…”