TY - JOUR
T1 - Glycoengineering human neural stem cells (hNSCs) for adhesion improvement using a novel thiol-modified N-acetylmannosamine (ManNAc) analog
AU - Du, Jian
AU - Liu, Xiao
AU - Yarema, Kevin J.
AU - Jia, Xiaofeng
N1 - Funding Information:
The work was supported by R01NS110387 and R01NS117102 (both to X. Jia) and R01EB005692 (to K. J. Yarema) from the National Institutes of Health (NIH). XJ and JD were partially supported by Maryland Stem Cell Research Fund , USA ( 2020-MSCRFD-5384 ) (to XJ). We acknowledge the partial support (to XJ) from the University of Maryland Baltimore , Institute for Clinical & Translational Research (ICTR) with the National Center for Advancing Translational Sciences (NCATS) CTSA grant number IULITR003098 .
Publisher Copyright:
© 2022
PY - 2022/3
Y1 - 2022/3
N2 - This study sets the stage for the therapeutic use of Ac5ManNTProp, an N-acetylmannosamine (ManNAc) analog that installs thiol-modified sialoglycans onto the surfaces of human neural stem cells (hNSC). First, we compared hNSC adhesion to the extracellular matrix (ECM) proteins laminin, fibronectin, and collagen and found preferential adhesion and concomitant changes to cell morphology and cell spreading for Ac5ManNTProp-treated cells to laminin, compared to fibronectin where there was a modest response, and collagen where there was no observable increase. PCR array transcript analysis identified several classes of cell adhesion molecules that responded to combined Ac5ManNTProp treatment and hNSC adhesion to laminin. Of these, we focused on integrin α6β1 expression, which was most strongly upregulated in analog-treated cells incubated on laminin. We also characterized downstream responses including vinculin display as well as the phosphorylation of focal adhesion kinase (FAK) and extracellular signal-related kinase (ERK). In these experiments, Ac5ManNTProp more strongly induced all tested biological endpoints compared to Ac5ManNTGc, showing that the single methylene unit that structurally separates the two analogs finely tunes biological responses. Together, the concerted modulation of multiple pro-regenerative activities through Ac5ManNTProp treatment, in concert with crosstalk with ECM components, lays a foundation for using our metabolic glycoengineering approach to treat neurological disorders by favorably modulating endpoints that contribute to the viability of transplanted NSCs.
AB - This study sets the stage for the therapeutic use of Ac5ManNTProp, an N-acetylmannosamine (ManNAc) analog that installs thiol-modified sialoglycans onto the surfaces of human neural stem cells (hNSC). First, we compared hNSC adhesion to the extracellular matrix (ECM) proteins laminin, fibronectin, and collagen and found preferential adhesion and concomitant changes to cell morphology and cell spreading for Ac5ManNTProp-treated cells to laminin, compared to fibronectin where there was a modest response, and collagen where there was no observable increase. PCR array transcript analysis identified several classes of cell adhesion molecules that responded to combined Ac5ManNTProp treatment and hNSC adhesion to laminin. Of these, we focused on integrin α6β1 expression, which was most strongly upregulated in analog-treated cells incubated on laminin. We also characterized downstream responses including vinculin display as well as the phosphorylation of focal adhesion kinase (FAK) and extracellular signal-related kinase (ERK). In these experiments, Ac5ManNTProp more strongly induced all tested biological endpoints compared to Ac5ManNTGc, showing that the single methylene unit that structurally separates the two analogs finely tunes biological responses. Together, the concerted modulation of multiple pro-regenerative activities through Ac5ManNTProp treatment, in concert with crosstalk with ECM components, lays a foundation for using our metabolic glycoengineering approach to treat neurological disorders by favorably modulating endpoints that contribute to the viability of transplanted NSCs.
KW - Cell adhesion
KW - Human neural stem cells
KW - ManNAc analog
KW - Metabolic glycoengineering
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U2 - 10.1016/j.msec.2022.112675
DO - 10.1016/j.msec.2022.112675
M3 - Article
C2 - 35599100
AN - SCOPUS:85128849406
SN - 2772-9508
VL - 134
JO - Biomaterials Advances
JF - Biomaterials Advances
M1 - 112675
ER -