Size-Optimized Layered Double Hydroxide Nanoparticles Promote Neural Progenitor Cells Differentiation of Embryonic Stem Cells Through the Regulation of M⁶A Methylation

Abstract

Purpose

The committed differentiation fate regulation has been a difficult problem in the fields of stem cell research, evidence showed that nanomaterials could promote the differentiation of stem cells into specific cell types. Layered double hydroxide (LDH) nanoparticles possess the regulation function of stem cell fate, while the underlying mechanism needs to be investigated. In this study, the process of embryonic stem cells (ESCs) differentiate to neural progenitor cells (NPCs) by magnesium aluminum LDH (MgAl-LDH) was investigated.

Methods

MgAl-LDH with diameters of 30, 50, and 100 nm were synthesized and characterized, and their effects on the cytotoxicity and differentiation of NPCs were detected in vitro. Dot blot and MeRIP-qPCR were performed to detect the level of m⁶A RNA methylation in nanoparticles-treated cells.

Results

Our work displayed that LDH nanoparticles of three different sizes were biocompatible with NPCs, and the addition of MgAl-LDH could significantly promote the process of ESCs differentiate to NPCs. 100 nm LDH has a stronger effect on promoting NPCs differentiation compared to 30 nm and 50 nm LDH. In addition, dot blot results indicated that the enhanced NPCs differentiation by MgAl-LDH was closely related to m⁶A RNA methylation process, and the major modification enzyme in LDH controlled NPCs differentiation may be the m⁶A RNA methyltransferase METTL3. The upregulated METTL3 by LDH increased the m⁶A level of Sox1 mRNA, enhancing its stability.

Conclusion

This work reveals that MgAl-LDH nanoparticles can regulate the differentiation of ESCs into NPCs by increasing m⁶A RNA methylation modification of Sox1.

Keywords:

• mouse embryonic stem cell
• neural progenitor cells
• layered double hydroxide nanoparticles
• size-dependent
• differentiation
• m⁶A RNA methylation

Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (Grant No. 82225027, 82202702, 82202351, 82301550, 82271418, 82001308, 82202803, and 82271419), the National Key Research and Development Program (Grant No. 2021YFA1101301), the Fundamental Research Funds for the Central Universities (Grant No. 22120230292, 22120220555, and 22120240163), the China Postdoctoral Science Foudation (2023M742647), and Shanghai Rising-Star Program (Grant No. 22QA1408200).

Disclosure

The authors report no conflicts of interest in this work.