The RNA helicases DDX5 and DDX17 facilitate neural differentiation of human pluripotent stem cells NTERA2
Issued Date
2022-02-15
Resource Type
ISSN
00243205
eISSN
18790631
Scopus ID
2-s2.0-85122624327
Pubmed ID
35007564
Journal Title
Life Sciences
Volume
291
Rights Holder(s)
SCOPUS
Bibliographic Citation
Life Sciences Vol.291 (2022)
Suggested Citation
Suthapot P., Xiao T., Felsenfeld G., Hongeng S., Wongtrakoongate P. The RNA helicases DDX5 and DDX17 facilitate neural differentiation of human pluripotent stem cells NTERA2. Life Sciences Vol.291 (2022). doi:10.1016/j.lfs.2021.120298 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/83824
Title
The RNA helicases DDX5 and DDX17 facilitate neural differentiation of human pluripotent stem cells NTERA2
Other Contributor(s)
Abstract
Aims: Understanding human neurogenesis is critical toward regenerative medicine for neurodegeneration. However, little is known how neural differentiation is regulated by DEAD box-containing RNA helicases, which comprise a diverse class of RNA remodeling enzymes. Materials and methods: ChIP-seq was utilized to identify binding sites of DDX5 and DDX17 in both human pluripotent stem cell (hPSC) line NTERA2 and their retinoic acid-induced neural derivatives. RNA-seq was used to elucidate genes differentially expressed upon depletion of DDX5 and DDX17. Neurosphere assay, flow cytometry, and immunofluorescence staining were performed to test the effect of depletion of the two RNA helicases in neural differentiation. Key findings: We show here that expression of DDX5 and DDX17 is abundant throughout neural differentiation of NTERA2, and is mostly localized within the nucleus. The two RNA helicases occupy chromatin genome-wide at regions associated with neurogenesis-related genes in both hPSCs and their neural derivatives. Further, both DDX5 and DDX17 are mutually required for controlling transcriptional expression of these genes, but are not important for maintenance of stem cell state of hPSCs. In contrast, they facilitate early neural differentiation of hPSCs, generation of neurospheres from the stem cells, and transcriptional expression of key neurogenic transcription factors such as SOX1 and PAX6 during neural differentiation. Importantly, DDX5 and DDX17 are critical for differentiation of hPSCs toward NESTIN- and TUBB3-positive cells, which represent neural progenitors and mature neurons, respectively. Significance: Collectively, our findings suggest the role of DDX5 and DDX17 in transcriptional regulation of genes involved in neurogenesis, and hence in neural differentiation of hPSCs.
