Essential Alterations of Heparan Sulfate During the Differentiation of Embryonic Stem Cells to Sox1-Enhanced Green Fluorescent Protein-Expressing Neural Progenitor Cells

Johnson, Claire; Crawford, Brett E.; Stavridis, Marios P.; Dam, Gerdy ten; Wat, Annie L; Rushton, Graham; Ward, Christopher M.; Wilson, Valerie; Kuppevelt, Toin H. Van; Smith, Austin; Gallagher, John T.; Merry, Catherine L.R.

doi:10.1634/stemcells.2006-0445

Cited by 123 publications

(144 citation statements)

References 40 publications

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“…Recent reports have demonstrated that the HS structure changes as ESCs undergo differentiation and that specific HS epitopes appear on subpopulations of differentiated ESCs, suggesting that HS regulates ESC differentiation and cell lineage development (21,22,24). This presumption has been supported by the examination of ESCs lacking Ndst1/2 (N-deacetylase/N-sulfotransferase 1/2) or Ext1.…”

mentioning

confidence: 92%

Heparan Sulfate Is Required for Embryonic Stem Cells to Exit from Self-renewal

Kraushaar

Yamaguchi

Wang

2010

Journal of Biological Chemistry

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Pluripotent embryonic stem cells (ESCs) must select between alternative fates of self-renewal and lineage commitment at each division during continuous proliferation. Heparan sulfate (HS) is a highly sulfated polysaccharide and is present abundantly on the ESC surface. In this study, we investigated the role of HS in ESC self-renewal by examining Ext1 ؊/؊ ESCs that are deficient in HS. We found that Ext1 ؊/؊ ESCs retained their self-renewal potential but failed to transit from self-renewal to differentiation upon removal of leukemia inhibitory factor. Furthermore, we found that the aberrant cell fate commitment is caused by defects in fibroblast growth factor signaling, which directly retained high expression of the pluripotency gene Nanog in Ext1 ؊/؊ ESCs. Therefore, our studies identified and defined HS as a novel factor that controls ESC fate commitment and also delineates that HS facilitates fibroblast growth factor signaling, which, in turn, inhibits Nanog expression and commits ESCs to lineage differentiation.Embryonic stem cells (ESCs) 2 are derived from the inner cell mass of the preimplantation blastocyst and can differentiate into numerous cell types representative of all three germ layers of the embryo, a property that is defined as pluripotency (1, 2). ESCs retain pluripotency through a process of self-renewal, which allows ESCs to proliferate infinitely as undifferentiated entities. These properties make ESCs a unique system to study early embryonic development and cell fate decisions and provide us with a promising source for cell replacement therapies (3-5). The regulatory network and molecular requirements for the maintenance of self-renewal have been under intense investigation and are now increasingly defined. However, the mechanisms by which ESCs exit the self-renewing state and initiate differentiation are still poorly understood. For example, recent reports suggest that extrinsic signaling of fibroblast growth factors (FGFs) and intracellular factors, including the chromatin-associated protein UTF1 (undifferentiated embryonic cell transcription factor 1) and the nucleosome remodeling complex, NuRD, are instrumental in triggering the exit of ESCs from their self-renewal program and to commit to differentiation (6 -8).Heparan sulfate (HS) is a highly sulfated glycosaminoglycan molecule and is biosynthesized in the Golgi apparatus of cells (9, 10). The copolymerases EXT1 and EXT2 initiate HS biosynthesis by alternately adding glucuronic acid and Nacetylglucosamine residues to form HS precursors. Following chain elongation, N-deacetylase/N-sulfotransferases act on discrete regions of the HS precursors, replacing N-acetyl groups with N-sulfate and creating appropriate substrates for further modification reactions, including epimerization and O-sulfation. The modification reactions are incomplete and result in mature HS that is structurally highly heterogeneous and possesses the potential to interact with a large variety of protein ligands. In tissues, the HS chains covalently attach to core prote...

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mentioning

confidence: 92%

Heparan Sulfate Is Required for Embryonic Stem Cells to Exit from Self-renewal

Kraushaar

Yamaguchi

Wang

2010

Journal of Biological Chemistry

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“…Following induction of spontaneous differentiation, Ext1 Ϫ/Ϫ ESCs showed a delay or absence of both endoderm and mesoderm markers (1). Furthermore, differentiation of Ext1 Ϫ/Ϫ ESCs into neuroectodermal precursors is defective (3). Recently, ESCs have been derived from Ext1 flox/flox embryos and following LIF withdrawal these ESCs showed a complete absence of markers of differentiation (4).…”

Section: Ext1mentioning

confidence: 99%

Influencing Hematopoietic Differentiation of Mouse Embryonic Stem Cells using Soluble Heparin and Heparan Sulfate Saccharides

Holley

Pickford

Rushton

et al. 2011

Journal of Biological Chemistry

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Heparan sulfate proteoglycans (HSPG) encompass some of the most abundant macromolecules on the surface of almost every cell type. Heparan sulfate (HS) chains provide a key interaction surface for the binding of numerous proteins such as growth factors and morphogens, helping to define the ability of a cell to respond selectively to environmental cues. The specificity of HSprotein interactions are governed predominantly by the order and positioning of sulfate groups, with distinct cell types expressing unique sets of HS epitopes. Embryos deficient in HS-synthesis (Ext1 ؊/؊ ) exhibit pre-gastrulation lethality and lack recognizable organized mesoderm and extraembryonic tissues. Here we demonstrate that embryonic stem cells (ESCs) derived from Ext1 ؊/؊ embryos are unable to differentiate into hematopoietic lineages, instead retaining ESC marker expression throughout embryoid body (EB) culture. However hematopoietic differentiation can be restored by the addition of soluble heparin. Consistent with specific size and composition requirements for HS:growth factor signaling, chains measuring at least 12 saccharides were required for partial rescue of hematopoiesis with longer chains (18 saccharides or more) required for complete rescue. Critically N-and 6-O-sulfate groups were essential for rescue. Heparin addition restored the activity of multiple signaling pathways including bone morphogenic protein (BMP) with activation of phosphoSMADs re-established by the addition of heparin. Heparin addition to wild-type cultures also altered the outcome of differentiation, promoting hematopoiesis at low concentrations, yet inhibiting blood formation at high concentrations. Thus altering the levels of HS and HS sulfation within differentiating ESC cultures provides an attractive and accessible mechanism for influencing cell fate.Cells have adopted elaborate systems to identify growth factor and morphogen cues in order to elicit an appropriate response throughout embryonic development and adulthood. A key component of this process is the sugar polymer heparan sulfate (HS), 2 which coats the outer surface of almost every cell within the body enabling the selective detection of environmental signals. HS polymerization is catalyzed by a heterodimer consisting of EXT1 and EXT2 enzymes resulting in the addition of alternating glucuronic acid (GlcA) and N-acetylglucosamine (GlcNAc) residues. This backbone is then enzymatically modified by de-N-acetylation and N-sulfation of selected GlcNAc residues to form N-sulfoglucosamine (GlcNS), epimerization of a subset of GlcA residues to iduronic acid (IdoUA) and the addition of sulfate groups at the 2-O-position of GlcA/IdoUA and 6-O-and rarely 3-O-position of GlcNS/GlcNAc. Modification is highly regulated and not complete, forming a cell-type specific pattern of highly sulfated domains (S-domains) interspersed by unmodified regions with the length, position and patterning of sulfate groups ultimately determining HS:ligand binding. The related GAG heparin is virtually fully sulfated along its l...

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“…Indeed, in EXT1 or EXT2-deficient mice, On the other hand, other groups reported that EXT1-deficient ES cells, which have no HS chains, could not differentiate (36,(40)(41)(42). In the EXT1-deficient ES cells, inhibition of Nanog expression via FGF4 signaling was eliminated because FGF4 signaling, the differentiation trigger, could not work with deficient HS chains.…”

Section: The Function Of the Hs Chain In Mouse Es Cellsmentioning

confidence: 99%

“…Sulfation pattern in HS is formed by several sulfotransferases and changes during differentiation of mouse ES cells (36). This finding was achieved by phage display antibodies that recognize specific sulfation structures of HS (36,48).…”

Section: F the Function Of Sulfation Pattern Of Hs In Mouse Es Cellsmentioning

confidence: 99%