Ex vivo glycan engineering of cd44 programs human multipotent mesenchymal stromal cell trafficking to bone

ABSTRACT The capacity to direct migration ('homing') of blood-borne cells to a predetermined anatomic compartment is vital to stem cell–based tissue engineering and other adoptive

cellular therapies. Although multipotent mesenchymal stromal cells (MSCs, also termed 'mesenchymal stem cells') hold the potential for curing generalized skeletal diseases, their

clinical effectiveness is constrained by the poor osteotropism of infused MSCs (refs. 1–3). Cellular recruitment to bone occurs within specialized marrow vessels that constitutively express

vascular E-selectin4,5, a lectin that recognizes sialofucosylated determinants on its various ligands. We show here that human MSCs do not express E-selectin ligands, but express a CD44

glycoform bearing α-2,3-sialyl modifications. Using an α-1,3-fucosyltransferase preparation and enzymatic conditions specifically designed for treating live cells, we converted the native

CD44 glycoform on MSCs into hematopoietic cell E-selectin/L-selectin ligand (HCELL)6, which conferred potent E-selectin binding without effects on cell viability or multipotency. Real-time

intravital microscopy in immunocompromised (NOD/SCID) mice showed that intravenously infused HCELL+ MSCs infiltrated marrow within hours of infusion, with ensuing rare foci of endosteally

localized cells and human osteoid generation. These findings establish that the HCELL glycoform of CD44 confers tropism to bone and unveil a readily translatable roadmap for programming

cellular trafficking by chemical engineering of glycans on a distinct membrane glycoprotein. Access through your institution Buy or subscribe This is a preview of subscription content,

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STROMAL CELLS WITH POLYVALENT ANTIBODIES TO ENHANCE TARGETING EFFICIENCY Article Open access 19 September 2023 ENFORCED MESENCHYMAL STEM CELL TISSUE COLONIZATION COUNTERACTS IMMUNOPATHOLOGY

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_Circ. Res._ 91, 907–914 (2002). Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS We thank C.A. Knoblauch, L. Liu and J.Y. Lee for assistance in manuscript preparation and

for skilled technical support, as well as P.V. Hauschka for helpful discussion of the data. We are grateful to the staff of the Cell Processing Laboratory of the Bone Marrow Transplantation

Unit at the Massachusetts General Hospital and the Cell Manipulation Core Facility of Dana Farber Cancer Center for their assistance in procuring the bone marrow harvest filter sets. This

effort was supported by National Institutes of Health grants RO1 HL73714 (R.S.), RO1 HL60528 (R.S.) and Massachusetts General Hospital Wellman Center Advanced Microscopy startup fund

(C.P.L.). This report is dedicated to the memory of Dr. Harvey R. Colten. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Departments of Dermatology and Medicine, Brigham and Women's

Hospital and Harvard Skin Disease Research Center, Harvard Medical School, Boston, 02115, Massachusetts, USA Robert Sackstein, Jasmeen S Merzaban, Derek W Cain & Nilesh M Dagia *

Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, 02115, Massachusetts, USA Robert Sackstein * Wellman Center for Photomedicine, Massachusetts

General Hospital, Harvard Medical School, Boston, 02115, Massachusetts, USA Joel A Spencer & Charles P Lin * Research Specialties, Sigma-Aldrich, Buchs, CH-9470, Switzerland Roland

Wohlgemuth Authors * Robert Sackstein View author publications You can also search for this author inPubMed Google Scholar * Jasmeen S Merzaban View author publications You can also search

for this author inPubMed Google Scholar * Derek W Cain View author publications You can also search for this author inPubMed Google Scholar * Nilesh M Dagia View author publications You can

also search for this author inPubMed Google Scholar * Joel A Spencer View author publications You can also search for this author inPubMed Google Scholar * Charles P Lin View author

publications You can also search for this author inPubMed Google Scholar * Roland Wohlgemuth View author publications You can also search for this author inPubMed Google Scholar

CONTRIBUTIONS R.S. conceived the study and reagents, created hybridomas, developed the SACK-1 mAb and the conditions for surface fucosylation of live cells, performed experiments and

supervised all research, wrote the manuscript and funded the research; J.S.M., D.W.C. and N.M.D. performed cell culture, biochemical studies and adhesion assays; J.A.S. and C.P.L. performed

intravital microscopy and C.P.L. partially funded the research; R.W. synthesized fucosyltransferase. CORRESPONDING AUTHOR Correspondence to Robert Sackstein. SUPPLEMENTARY INFORMATION

SUPPLEMENTARY TEXT AND FIGURES Supplementary Figs. 1–6 and Supplementary Methods (PDF 5151 kb) SUPPLEMENTARY MOVIE 1 This video segment shows a marrow sinusoidal endothelial bed within the

first minute after injection of FTVI-treated, HCELL-expressing MSCs (bright cells). Note evident rolling interactions and firm adherence of MSCs onto marrow sinusoidal endothelium. (MP4 3000

kb) SUPPLEMENTARY MOVIE 2 This video segment shows a marrow sinusoidal endothelial bed withing the first minute after injection of FTVI-Sialidase MSCs. Compared to HCELL+ MSCs shown in

Video 1, FTVI-Sialidase MSCs show minimal binding interactions with marrow sinusoidal endothelium. (MP4 3100 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS

ARTICLE Sackstein, R., Merzaban, J., Cain, D. _et al._ _Ex vivo_ glycan engineering of CD44 programs human multipotent mesenchymal stromal cell trafficking to bone. _Nat Med_ 14, 181–187

(2008). https://doi.org/10.1038/nm1703 Download citation * Received: 22 October 2007 * Accepted: 05 December 2007 * Published: 13 January 2008 * Issue Date: February 2008 * DOI:

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