Glycoengineering of E-Selectin Ligands by Intracellular versus Extracellular Fucosylation Differentially Affects Osteotropism of Human Mesenchymal Stem Cells

被引：46

作者：

Dykstra, Brad ^{[1
,2
,3
]}

Lee, Jungmin ^{[4
,5
]}

Mortensen, Luke J. ^{[6
,7
]}

Yu, Haixiao ^{[8
]}

Wu, Zhengliang L. ^{[8
]}

Lin, Charles P. ^{[9
,10
]}

Rossi, Derrick J. ^{[4
,5
]}

Sackstein, Robert ^{[1
,2
,3
,11
]}

机构：

[1] Brigham & Womens Hosp, Dept Dermatol, Boston, MA 02115 USA

[2] Brigham & Womens Hosp, Harvard Skin Dis Res Ctr, Boston, MA 02115 USA

[3] Boston Childrens Hosp, Program Excellence Glycosci, Boston, MA USA

[4] Boston Childrens Hosp, Program Cellular & Mol Med, Div Hematol Oncol, Boston, MA USA

[5] Harvard Univ, Dept Stem Cell & Regenerat Biol, Cambridge, MA 02138 USA

[6] Univ Georgia, Regenerat Biosci Ctr, Rhodes Ctr Anim & Dairy Sci, Athens, GA 30602 USA

[7] Univ Georgia, Coll Engn, Athens, GA 30602 USA

[8] R&D Syst Inc, Biotechne, Minneapolis, MN USA

[9] Massachusetts Gen Hosp, Adv Microscopy Program, Ctr Syst Biol, Boston, MA 02114 USA

[10] Massachusetts Gen Hosp, Wellman Ctr Photomed, Boston, MA 02114 USA

[11] Harvard Med Sch, Brigham & Womens Hosp, Dept Med, Boston, MA USA

来源：

STEM CELLS | 2016年 / 34卷 / 10期

关键词：

Mesenchymal stromal cell; HCELL; GPS; E-selectin; sialyl Lewis X; Fucosyltransferase; Exofucosylation; Modified mRNA; Intravital microscopy; MODIFIED MESSENGER-RNA; STROMAL CELLS; BONE-MARROW; ENHANCES ENGRAFTMENT; P-SELECTIN; IN-VIVO; EXPRESSION; CXCR4; TRANSPLANTATION; PLURIPOTENCY;

D O I：

10.1002/stem.2435

中图分类号：

Q813 [细胞工程];

学科分类号：

摘要：

Human mesenchymal stem cells (MSCs) hold great promise in cellular therapeutics for skeletal diseases but lack expression of E-selectin ligands that direct homing of blood-borne cells to bone marrow. Previously, we described a method to engineer E-selectin ligands on the MSC surface by exofucosylating cells with fucosyltransferase VI (FTVI) and its donor sugar, GDP-Fucose, enforcing transient surface expression of the potent E-selectin ligand HCELL with resultant enhanced osteotropism of intravenously administered cells. Here, we sought to determine whether E-selectin ligands created via FTVI-exofucosylation are distinct in identity and function to those created by FTVI expressed intracellularly. To this end, we introduced synthetic modified mRNA encoding FTVI (FUT6-modRNA) into human MSCs. FTVI-exofucosylation (i.e., extracellular fucosylation) and FUT6-modRNA transfection (i.e., intracellular fucosylation) produced similar peak increases in cell surface E-selectin ligand levels, and shear-based functional assays showed comparable increases in tethering/rolling on human endothelial cells expressing E-selectin. However, biochemical analyses revealed that intracellular fucosylation induced expression of both intracellular and cell surface E-selectin ligands and also induced a more sustained expression of E-selectin ligands compared to extracellular fucosylation. Notably, live imaging studies to assess homing of human MSC to mouse calvarium revealed more osteotropism following intravenous administration of intracellularly-fucosylated cells compared to extracellularly-fucosylated cells. This study represents the first direct analysis of E-selectin ligand expression programmed on human MSCs by FTVI-mediated intracellular versus extracellular fucosylation. The observed differential biologic effects of FTVI activity in these two contexts may yield new strategies for improving the efficacy of human MSCs in clinical applications. Stem Cells2016;34:2501-2511

引用

页码：2501 / 2511

页数：11

共 32 条

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