SpringerLink
Log in

Histone H2A deubiquitinase BAP1 is essential for endothelial cell differentiation from human pluripotent stem cells

  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Abstract

Polycomb group proteins (PcGs) add repressive post translational histone modifications such as H2AK119ub1, and histone H2A deubiquitinases remove it. Mice lacking histone H2A deubiquitinases such as Usp16 and Bap1 die in embryonic stage, while mice lacking Usp3, Mysm1, Usp12, and Usp21 have been shown to be deficient in hematopoietic lineage differentiation, cell cycle regulation, and DNA repair. Thus, it is likely that histone deubiquitinases may also be required for human endothelial cell differentiation; however, there are no reports about the role of histone H2A deubiquitinase BAP1 in human endothelial cell development. We differentiated human pluripotent stem cells into the endothelial lineage which expressed stable inducible shRNA against BAP1. Our results show that BAP1 is required for human endothelial cell differentiation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1.
Figure 2.
Figure 3.
Figure 4.

Data availability

The data that support the findings of the study are available with the corresponding author upon request.

References

  • Bauer M, Trupke J, Ringrose L (2016) The quest for mammalian Polycomb response elements: are we there yet? Chromosoma 125(3):471–496

    Article  CAS  PubMed  Google Scholar 

  • Belle JI, Nijnik A (2014) H2A-DUBbing the mammalian epigenome: expanding frontiers for histone H2A deubiquitinating enzymes in cell biology and physiology. Int J Biochem Cell Biol 50(1):161–174

    Article  CAS  PubMed  Google Scholar 

  • de Napoles M, Mermoud JE, Wakao R, Tang YA, Endoh M, Appanah R, Nesterova TB, Silva J, Otte AP, Vidal M, Koseki H, Brockdorff N (2004) Polycomb group proteins Ring1A/B link ubiquitylation of histone H2A to heritable gene silencing and X inactivation. Dev Cell 7(5):663–676

    Article  PubMed  Google Scholar 

  • Desai D, Khanna A, Pethe P (2020) PRC1 catalytic unit RING1B regulates early neural differentiation of human pluripotent stem cells. Exp Cell Res 396(1):112294

    Article  CAS  PubMed  Google Scholar 

  • Dey A, Seshasayee D, Noubade R, French DM, Liu J, Chaurushiya MS, Kirkpatrick DS, Pham VC, Lill JR, Bakalarski CE, Wu J, Phu L, Katavolos P, LaFave LM, Abdel-Wahab O, Modrusan Z, Seshagiri S, Dong K, Lin Z et al (2012) Loss of the tumor suppressor BAP1 causes myeloid transformation. Science (New York, N.Y.) 337(6101):1541–1546

    Article  CAS  PubMed  Google Scholar 

  • Ding X, Lin Q, Ensenat-Waser R, Rose-John S, Zenke M (2012) Polycomb group protein Bmi1 promotes hematopoietic cell development from embryonic stem cells. Stem Cells Dev 21(1):121–132

    Article  CAS  PubMed  Google Scholar 

  • Dumasia NP, Khanna AP, Pethe PS (2021) Sonic hedgehog signals hinder the transcriptional network necessary for pancreatic endoderm formation from human embryonic stem cells. Genes Cells 26(5):282–297

    Article  CAS  PubMed  Google Scholar 

  • Gibney ER, Nolan CM (2010) Epigenetics and gene expression. Heredity 105(1):4–13

    Article  CAS  PubMed  Google Scholar 

  • Golbabapour S, Majid NA, Hassandarvish P, Hajrezaie M, Abdulla MA, Hadi AH (2013) Gene silencing and Polycomb group proteins: an overview of their structure, mechanisms and phylogenetics. Omics : J Integ Biol 17(6):283–296

    Article  CAS  Google Scholar 

  • Guibentif C, Rönn RE, Böiers C, Lang S, Saxena S, Soneji S, Enver T, Karlsson G, Woods NB (2017) Single-cell analysis identifies distinct stages of human endothelial-to-hematopoietic transition. Cell Rep 19(1):10–19

    Article  CAS  PubMed  Google Scholar 

  • Jeong J, Jung I, Kim JH, Jeon S, Hyeon DY, Min H, Kang B, Nah J, Hwang D, Um SJ, Ko M, Seong RH (2022) BAP1 shapes the bone marrow niche for lymphopoiesis by fine-tuning epigenetic profiles in endosteal mesenchymal stromal cells. Cell Death Differ 29(11):2151–2162

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Koutelou E, Wang L, Schibler AC, Chao HP, Kuang X, Lin K, Lu Y, Shen J, Jeter CR, Salinger A, Wilson M, Chen YC, Atanassov BS, Tang DG, Dent SYR (2019) USP22 controls multiple signaling pathways that are essential for vasculature formation in the mouse placenta. Development 146(4):dev174037

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kumar N, Hinduja I, Nagvenkar P, Pillai L, Zaveri K, Mukadam L, Telang J, Desai S, Mangoli V, Mangoli R, Padgaonkar S, Kaur G, Puri C, Bhartiya D (2009) Derivation and characterization of two genetically unique human embryonic stem cell lines on in-house-derived human feeders. Stem Cells Dev 18(3):435–445

    Article  CAS  PubMed  Google Scholar 

  • Kuznetsov JN, Aguero TH, Owens DA, Kurtenbach S, Field MG, Durante MA, Rodriguez DA, King ML, Harbour JW (2019) BAP1 regulates epigenetic switch from pluripotency to differentiation in developmental lineages giving rise to BAP1-mutant cancers. Sci Adv 5(9):eaax1738

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li R, Huang D, Zhao Y, Yuan Y, Sun X, Dai Z, Huo D, Liu X, Helin K, Li MJ, Wu X (2023) PR-DUB safeguards Polycomb repression through H2AK119ub1 restriction. Cell proliferation 56(10):e13457

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Masclef L, Ahmed O, Estavoyer B, Larrivée B, Labrecque N, Nijnik A, Affar EB (2021) Roles and mechanisms of BAP1 deubiquitinase in tumor suppression. Cell Death Differ 28(2):606–625

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Perez-Garcia V, Lea G, Lopez-Jimenez P, Okkenhaug H, Burton GJ, Moffett A, Turco MY, Hemberger M (2021) BAP1/ASXL complex modulation regulates epithelial-mesenchymal transition during trophoblast differentiation and invasion. eLife 10:e63254

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pethe P, Noel VS, Kale V (2021) Deterministic role of sonic hedgehog signalling pathway in specification of hemogenic versus endocardiogenic endothelium from differentiated human embryonic stem cells. Cells Dev 166:203685

    Article  CAS  PubMed  Google Scholar 

  • Psaltis PJ, Harbuzariu A, Delacroix S, Holroyd EW, Simari RD (2011) Resident vascular progenitor cells--diverse origins, phenotype, and function. J Cardio Transl Res 4(2):161–176

    Article  Google Scholar 

  • Shin JH, Seo BG, Lee IW, Kim HJ, Seo EC, Lee KM, Jeon SB, Baek SK, Kim TS, Lee JH, Choi JW, Hwangbo C, Lee JH (2022) Functional characterization of endothelial cells differentiated from porcine epiblast stem cells. Cells 11(9):1524

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Simmons DL, Satterthwaite AB, Tenen DG, Seed B (1992) Molecular cloning of a cDNA encoding CD34, a sialomucin of human hematopoietic stem cells. J Immunol 148(1):267–271

    Article  CAS  PubMed  Google Scholar 

  • Takano J, Ito S, Dong Y, Sharif J, Nakajima-Takagi Y, Umeyama T, Han YW, Isono K, Kondo T, Iizuka Y, Miyai T, Koseki Y, Ikegaya M, Sakihara M, Nakayama M, Ohara O, Hasegawa Y, Hashimoto K, Arner E et al (2022) PCGF1-PRC1 links chromatin repression with DNA replication during hematopoietic cell lineage commitment. Nat Commun 13(1):7159

    Article  PubMed  PubMed Central  Google Scholar 

  • Tamburri S, Conway E, Pasini D (2022) Polycomb-dependent histone H2A ubiquitination links developmental disorders with cancer. Trends Genetics : TIG 38(4):333–352

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Langlais D, Nijnik A (2023) Histone H2A deubiquitinases in the transcriptional programs of development and hematopoiesis: a consolidated analysis. The Int J Biochem Cell Biol 157:106384

    Article  CAS  PubMed  Google Scholar 

  • Zhao S, Feng S, Tian Y, Wen Z (2022) Hemogenic and aortic endothelium arise from a common hemogenic angioblast precursor and are specified by the Etv2 dosage. Proc Natl Acad Sci USA 119(13):e2119051119

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhen F, Lan Y, Yan B, Zhang W, Wen Z (2013) Hemogenic endothelium specification and hematopoietic stem cell maintenance employ distinct Scl isoforms. Development 140(19):3977–3985

    Article  CAS  PubMed  Google Scholar 

  • Zhu Q, Gao P, Tober J, Bennett L, Chen C, Uzun Y, Li Y, Howell ED, Mumau M, Yu W, He B, Speck NA, Tan K (2020) Developmental trajectory of prehematopoietic stem cell formation from endothelium. Blood 136(7):845–856

    Article  PubMed  PubMed Central  Google Scholar 

  • Zovein AC, Hofmann JJ, Lynch M, French WJ, Turlo KA, Yang Y, Becker MS, Zanetta L, Dejana E, Gasson JC, Tallquist MD, Iruela-Arispe ML (2008) Fate tracing reveals the endothelial origin of hematopoietic stem cells. Cell Stem Cell 3(6):625–636

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Funding

The work was partly funded by the Department of Biotechnology (DBT), Govt of India, BT/PR41465/MED/31/435/2020.

Author information

Authors and Affiliations

  1. Symbiosis Centre for Stem Cell Research (SCSCR), Symbiosis International (Deemed University), Pune, India

    Shruti Shastry, Dharitree Samal & Prasad Pethe

  2. Worcester Polytechnic Institute (WPI), Boston, USA

    Shruti Shastry

Authors
  1. Shruti Shastry
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dharitree Samal
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Prasad Pethe
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Prasad Pethe.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Supplementary information

ESM 1

(JPG 166 kb)

ESM 2

Fig S1 shows western blot of BAP1 in all 3 biological replicates along with loading control β-ACTIN. Fig S2 represents the effect of BAP1 knockdown on the cell size of shRNA scrambled (control), shRNA1 BAP1 and shRNA2 BAP1. Fig S3 shows western blot for CD31 in scrambled, shRNA1 and shRNA2 along with its densitometric analysis (Fig S4). Fig S6 depicts western blot of RUNX2 in scrambled, shRNA1 and shRNA2 along with its densitometric analysis (Fig S6). Fig S7 shows the negative control panel for immunofluorescence for figure 1B. (DOCX 21380 kb)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shastry, S., Samal, D. & Pethe, P. Histone H2A deubiquitinase BAP1 is essential for endothelial cell differentiation from human pluripotent stem cells. In Vitro Cell.Dev.Biol.-Animal (2024). https://doi.org/10.1007/s11626-024-00935-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11626-024-00935-x

Keywords

Search

Navigation