1. B3GALT5 knockout alters gycosphingolipid profile and facilitates transition to human naïve pluripotency.
- Author
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Lin RJ, Kuo MW, Yang BC, Tsai HH, Chen K, Huang JR, Lee YS, Yu AL, and Yu J
- Subjects
- CRISPR-Cas Systems genetics, Cell Line, Embryonic Stem Cells, Galactosyltransferases genetics, Gene Knockdown Techniques, Humans, Cell Differentiation, Galactosyltransferases metabolism, Glycosphingolipids metabolism, Pluripotent Stem Cells metabolism, Stage-Specific Embryonic Antigens metabolism
- Abstract
Conversion of human pluripotent stem cells from primed to naïve state is accompanied by altered transcriptome and methylome, but glycosphingolipid (GSL) profiles in naïve human embryonic stem cells (hESCs) have not been systematically characterized. Here we showed a switch from globo-(SSEA-3, SSEA-4, and Globo H) and lacto-series (fucosyl-Lc4Cer) to neolacto-series GSLs (SSEA-1 and H type 2 antigen), along with marked down-regulation of β-1,3-galactosyltransferase (B3GALT5) upon conversion to naïve state. CRISPR/Cas9-generated B3GALT5- knockout (KO) hESCs displayed an altered GSL profile, increased cloning efficiency and intracellular Ca
2+ , reminiscent of the naïve state, while retaining differentiation ability. The altered GSLs could be rescued through overexpression of B3GALT5. B3GALT5- KO cells cultured with 2iLAF exhibited naïve-like transcriptome, global DNA hypomethylation, and X-chromosome reactivation. In addition, B3GALT5 -KO rendered hESCs more resistant to calcium chelator in blocking entry into naïve state. Thus, loss of B3GALT5 induces a distinctive state of hESCs displaying unique GSL profiling with expression of neolacto-glycans, increased Ca2+ , and conducive for transition to naïve pluripotency., Competing Interests: The authors declare no competing interest., (Copyright © 2020 the Author(s). Published by PNAS.)- Published
- 2020
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