Snap29 Is Dispensable for Self-Renewal Maintenance but Required for Proper Differentiation of Mouse Embryonic Stem Cells

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2023 Jan 8

PMID 36614195

Authors

Yumei Jia

Zhaoyuan Guo

Jiahao Zhu

Guanyu Qin

Wenwen Sun

Yu Yin

Haiying Wang

Renpeng Guo

Affiliations

Soon will be listed here.

Abstract

Pluripotent embryonic stem cells (ESCs) can self-renew indefinitely and are able to differentiate into all three embryonic germ layers. Synaptosomal-associated protein 29 (Snap29) is implicated in numerous intracellular membrane trafficking pathways, including autophagy, which is involved in the maintenance of ESC pluripotency. However, the function of Snap29 in the self-renewal and differentiation of ESCs remains elusive. Here, we show that depletion via CRISPR/Cas does not impair the self-renewal and expression of pluripotency-associated factors in mouse ESCs. However, deficiency enhances the differentiation of ESCs into cardiomyocytes, as indicated by heart-like beating cells. Furthermore, transcriptome analysis reveals that depletion significantly decreased the expression of numerous genes required for germ layer differentiation. Interestingly, deficiency does not cause autophagy blockage in ESCs, which might be rescued by the SNAP family member Snap47. Our data show that Snap29 is dispensable for self-renewal maintenance, but required for the proper differentiation of mouse ESCs.

References

Morelli E, Ginefra P, Mastrodonato V, Beznoussenko G, Rusten T, Bilder D . Multiple functions of the SNARE protein Snap29 in autophagy, endocytic, and exocytic trafficking during epithelial formation in Drosophila. Autophagy. 2015; 10(12):2251-68. PMC: 4502674. DOI: 10.4161/15548627.2014.981913. View

Martens M, Edelkamp J, Seebode C, Schafer M, Stahlke S, Krohn S . Generation and Characterization of a CRISPR/Cas9-Mediated Knockout in Human Fibroblasts. Int J Mol Sci. 2021; 22(10). PMC: 8157373. DOI: 10.3390/ijms22105293. View

Li H, Wang R, Yu Z, Shi R, Zhang J, Gao S . Tumor Necrosis Factor α Reduces SNAP29 Dependent Autolysosome Formation to Increase Prion Protein Level and Promote Tumor Cell Migration. Virol Sin. 2020; 36(3):458-475. PMC: 8257879. DOI: 10.1007/s12250-020-00320-4. View

Boyer L, Lee T, Cole M, Johnstone S, Levine S, Zucker J . Core transcriptional regulatory circuitry in human embryonic stem cells. Cell. 2005; 122(6):947-56. PMC: 3006442. DOI: 10.1016/j.cell.2005.08.020. View

Zhang Q, Mao J, Zhang X, Fu H, Xia S, Yin Z . Role of Jnk1 in development of neural precursors revealed by iPSC modeling. Oncotarget. 2016; 7(38):60919-60928. PMC: 5308626. DOI: 10.18632/oncotarget.11377. View

Feyen D, McKeithan W, Bruyneel A, Spiering S, Hormann L, Ulmer B . Metabolic Maturation Media Improve Physiological Function of Human iPSC-Derived Cardiomyocytes. Cell Rep. 2020; 32(3):107925. PMC: 7437654. DOI: 10.1016/j.celrep.2020.107925. View

Rapaport D, Lugassy Y, Sprecher E, Horowitz M . Loss of SNAP29 impairs endocytic recycling and cell motility. PLoS One. 2010; 5(3):e9759. PMC: 2841205. DOI: 10.1371/journal.pone.0009759. View

Hwang Y, Chung B, Ortmann D, Hattori N, Moeller H, Khademhosseini A . Microwell-mediated control of embryoid body size regulates embryonic stem cell fate via differential expression of WNT5a and WNT11. Proc Natl Acad Sci U S A. 2009; 106(40):16978-83. PMC: 2761314. DOI: 10.1073/pnas.0905550106. View

Liu K, Cao J, Shi X, Wang L, Zhao T . Cellular metabolism and homeostasis in pluripotency regulation. Protein Cell. 2020; 11(9):630-640. PMC: 7452966. DOI: 10.1007/s13238-020-00755-1. View

10.

Schiller S, Seebode C, Wieser G, Goebbels S, Mobius W, Horowitz M . Establishment of Two Mouse Models for CEDNIK Syndrome Reveals the Pivotal Role of SNAP29 in Epidermal Differentiation. J Invest Dermatol. 2016; 136(3):672-679. DOI: 10.1016/j.jid.2015.12.020. View

11.

Takats S, Nagy P, Varga A, Pircs K, Karpati M, Varga K . Autophagosomal Syntaxin17-dependent lysosomal degradation maintains neuronal function in Drosophila. J Cell Biol. 2013; 201(4):531-9. PMC: 3653357. DOI: 10.1083/jcb.201211160. View

12.

Orkin S, Wang J, Kim J, Chu J, Rao S, Theunissen T . The transcriptional network controlling pluripotency in ES cells. Cold Spring Harb Symp Quant Biol. 2009; 73:195-202. DOI: 10.1101/sqb.2008.72.001. View

13.

Kwon G, Fraser S, Eakin G, Mangano M, Isern J, Sahr K . Tg(Afp-GFP) expression marks primitive and definitive endoderm lineages during mouse development. Dev Dyn. 2006; 235(9):2549-58. PMC: 1850385. DOI: 10.1002/dvdy.20843. View

14.

Guo R, Ye X, Yang J, Zhou Z, Tian C, Wang H . Feeders facilitate telomere maintenance and chromosomal stability of embryonic stem cells. Nat Commun. 2018; 9(1):2620. PMC: 6033898. DOI: 10.1038/s41467-018-05038-2. View

15.

Mohamud Y, Shi J, Qu J, Poon T, Xue Y, Deng H . Enteroviral Infection Inhibits Autophagic Flux via Disruption of the SNARE Complex to Enhance Viral Replication. Cell Rep. 2018; 22(12):3292-3303. DOI: 10.1016/j.celrep.2018.02.090. View

16.

Liu N, Yin Y, Wang H, Zhou Z, Sheng X, Fu H . Telomere dysfunction impairs epidermal stem cell specification and differentiation by disrupting BMP/pSmad/P63 signaling. PLoS Genet. 2019; 15(9):e1008368. PMC: 6760834. DOI: 10.1371/journal.pgen.1008368. View

17.

Takats S, Glatz G, Szenci G, Boda A, Horvath G, Hegedus K . Non-canonical role of the SNARE protein Ykt6 in autophagosome-lysosome fusion. PLoS Genet. 2018; 14(4):e1007359. PMC: 5937789. DOI: 10.1371/journal.pgen.1007359. View

18.

Mukund K, Subramaniam S . Skeletal muscle: A review of molecular structure and function, in health and disease. Wiley Interdiscip Rev Syst Biol Med. 2019; 12(1):e1462. PMC: 6916202. DOI: 10.1002/wsbm.1462. View

19.

Sheikh M, Emerald B, Ansari S . Stem cell fate determination through protein O-GlcNAcylation. J Biol Chem. 2020; 296:100035. PMC: 7948975. DOI: 10.1074/jbc.REV120.014915. View

20.

De Los Angeles A, Ferrari F, Xi R, Fujiwara Y, Benvenisty N, Deng H . Hallmarks of pluripotency. Nature. 2015; 525(7570):469-78. DOI: 10.1038/nature15515. View