Differentiation of HPSCs to Study PRC2 Role in Cell-Fate Specification and Neurodevelopment

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2023 May 22

PMID 37212999

Authors

Silvia Brocchetti

Paola Conforti

Affiliations

Soon will be listed here.

Abstract

Several studies highlighted the importance of the polycomb repressive complex 2 (PRC2) already at the beginning of development. Although the crucial function of PRC2 in regulating lineage commitment and cell-fate specification has been well-established, the in vitro study of the exact mechanisms for which H3K27me3 is indispensable for proper differentiation is still challenging. In this chapter, we report a well-established and reproducible differentiation protocol to generate striatal medium spiny neurons as a tool to explore PRC2 role in brain development.

References

Budday S, Steinmann P, Kuhl E . Physical biology of human brain development. Front Cell Neurosci. 2015; 9:257. PMC: 4495345. DOI: 10.3389/fncel.2015.00257. View

Rubenstein J, Shimamura K, Martinez S, Puelles L . Regionalization of the prosencephalic neural plate. Annu Rev Neurosci. 1998; 21:445-77. DOI: 10.1146/annurev.neuro.21.1.445. View

WINDLE W, CLEARWATERS P . Development of the nervous system. Prog Neurol Psychiatry. 2010; 1:18-25. View

Shi Y, Kirwan P, Livesey F . Directed differentiation of human pluripotent stem cells to cerebral cortex neurons and neural networks. Nat Protoc. 2012; 7(10):1836-46. DOI: 10.1038/nprot.2012.116. View

Boyer L, Plath K, Zeitlinger J, Brambrink T, Medeiros L, Lee T . Polycomb complexes repress developmental regulators in murine embryonic stem cells. Nature. 2006; 441(7091):349-53. DOI: 10.1038/nature04733. View

Lee T, Jenner R, Boyer L, Guenther M, Levine S, Kumar R . Control of developmental regulators by Polycomb in human embryonic stem cells. Cell. 2006; 125(2):301-13. PMC: 3773330. DOI: 10.1016/j.cell.2006.02.043. View

Erhardt S, Su I, Schneider R, Barton S, Bannister A, Perez-Burgos L . Consequences of the depletion of zygotic and embryonic enhancer of zeste 2 during preimplantation mouse development. Development. 2003; 130(18):4235-48. DOI: 10.1242/dev.00625. View

Pasini D, Bracken A, Jensen M, Denchi E, Helin K . Suz12 is essential for mouse development and for EZH2 histone methyltransferase activity. EMBO J. 2004; 23(20):4061-71. PMC: 524339. DOI: 10.1038/sj.emboj.7600402. View

Mager J, Montgomery N, Pardo-Manuel de Villena F, Magnuson T . Genome imprinting regulated by the mouse Polycomb group protein Eed. Nat Genet. 2003; 33(4):502-7. DOI: 10.1038/ng1125. View

10.

Shan Y, Liang Z, Xing Q, Zhang T, Wang B, Tian S . PRC2 specifies ectoderm lineages and maintains pluripotency in primed but not naïve ESCs. Nat Commun. 2017; 8(1):672. PMC: 5610324. DOI: 10.1038/s41467-017-00668-4. View

11.

Pasini D, Bracken A, Hansen J, Capillo M, Helin K . The polycomb group protein Suz12 is required for embryonic stem cell differentiation. Mol Cell Biol. 2007; 27(10):3769-79. PMC: 1899991. DOI: 10.1128/MCB.01432-06. View

12.

Sparmann A, van Lohuizen M . Polycomb silencers control cell fate, development and cancer. Nat Rev Cancer. 2006; 6(11):846-56. DOI: 10.1038/nrc1991. View

13.

Takeuchi T, Yamazaki Y, Tsuchiya R, Kondo S, MOTOYAMA J, Higashinakagawa T . Gene trap capture of a novel mouse gene, jumonji, required for neural tube formation. Genes Dev. 1995; 9(10):1211-22. DOI: 10.1101/gad.9.10.1211. View

14.

Volcik K, Zhu H, Finnell R, Shaw G, Canfield M, Lammer E . Evaluation of the jumonji gene and risk for spina bifida and congenital heart defects. Am J Med Genet A. 2004; 126A(2):215-7. DOI: 10.1002/ajmg.a.20574. View

15.

Pietersen A, van Lohuizen M . Stem cell regulation by polycomb repressors: postponing commitment. Curr Opin Cell Biol. 2008; 20(2):201-7. DOI: 10.1016/j.ceb.2008.01.004. View

16.

Bahmad H, Hadadeh O, Chamaa F, Cheaito K, Darwish B, Makkawi A . Modeling Human Neurological and Neurodegenerative Diseases: From Induced Pluripotent Stem Cells to Neuronal Differentiation and Its Applications in Neurotrauma. Front Mol Neurosci. 2017; 10:50. PMC: 5329035. DOI: 10.3389/fnmol.2017.00050. View

17.

Gaspard N, Vanderhaeghen P . From stem cells to neural networks: recent advances and perspectives for neurodevelopmental disorders. Dev Med Child Neurol. 2010; 53(1):13-7. DOI: 10.1111/j.1469-8749.2010.03827.x. View

18.

Biagioli M, Ferrari F, Mendenhall E, Zhang Y, Erdin S, Vijayvargia R . Htt CAG repeat expansion confers pleiotropic gains of mutant huntingtin function in chromatin regulation. Hum Mol Genet. 2015; 24(9):2442-57. PMC: 4383859. DOI: 10.1093/hmg/ddv006. View

19.

Seong I, Woda J, Song J, Lloret A, Abeyrathne P, Woo C . Huntingtin facilitates polycomb repressive complex 2. Hum Mol Genet. 2009; 19(4):573-83. PMC: 2807366. DOI: 10.1093/hmg/ddp524. View

20.

Reiner A, Albin R, Anderson K, DAMATO C, Penney J, Young A . Differential loss of striatal projection neurons in Huntington disease. Proc Natl Acad Sci U S A. 1988; 85(15):5733-7. PMC: 281835. DOI: 10.1073/pnas.85.15.5733. View