HCELL Expression on Murine MSC Licenses Pancreatotropism and Confers Durable Reversal of Autoimmune Diabetes in NOD Mice

Abstract

Type 1 diabetes (T1D) is an immune-mediated disease resulting in destruction of insulinproducing pancreatic beta cells. Mesenchymal stem cells (MSCs) possess potent immunomodulatory properties, garnering increasing attention as cellular therapy for T1D and other immunologic diseases. However, MSCs generally lack homing molecules, hindering their colonization at inflammatory sites following intravenous (IV) administration. Here, we analyzed whether enforced E-selectin ligand expression on murine MSCs could impact their effect in reversing hyperglycemia in nonobese diabetic (NOD) mice. Although murine MSCs natively do not express the E-selectin-binding determinant sialyl Lewis x (sLe x ), we found that fucosyltransferasemediated a(1,3)-exofucosylation of murine MSCs resulted in sLe x display uniquely on cell surface CD44 thereby creating hematopoietic cell E-/L-selectin ligand (HCELL), the E-selectin-binding glycoform of CD44. Following IV infusion into diabetic NOD mice, allogeneic HCELL 1 MSCs showed threefold greater peri-islet infiltrates compared to buffer-treated (i.e., HCELL 2 ) MSCs, with distribution in proximity to E-selectin-expressing microvessels. Exofucosylation had no effect on MSC immunosuppressive capacity in in vitro assays; however, although engraftment was temporary for both HCELL 1 and HCELL 2 MSCs, administration of HCELL 1 MSCs resulted in durable reversal of hyperglycemia, whereas only transient reversal was observed following administration of HCELL 2 MSCs. Notably, exofucosylation of MSCs generated from CD44 2/2 mice induced prominent membrane expression of sLe x , but IV administration of these MSCs into hyperglycemic NOD mice showed no enhanced pancreatotropism or reversal of hyperglycemia. These findings provide evidence that glycan engineering to enforce HCELL expression boosts trafficking of infused MSCs to pancreatic islets of NOD mice and substantially improves their efficacy in reversing autoimmune diabetes. STEM CELLS 2013;33:1523-1531

Imaging specific cellular glycan structures using glycosyltransferases via click chemistry

“…indicates that there are sLN on C3H10T1/2 cells, which is consistent with previous findings that both mouse and human mesenchymal stem cells can be (1,3)-exofucosylated to enforce sLeX expression (Abdi, R., Moore, R., et al 2015, Dykstra, B., Lee, J., et al 2016). The staining observed using GCNT1 indicates that C3H10T1/2 cells express core-1 O-glycan or T antigen.…”

E-Selectin Ligands in the Human Mononuclear Phagocyte System: Implications for Infection, Inflammation, and Immunotherapy

“…Inflammatory bowel disease ( 199 – 202 ), multiple sclerosis ( 203 , 204 ), rheumatoid arthritis ( 205 – 207 ), and type 1 diabetes ( 208 , 209 ) are other examples of inflammatory diseases in which upregulated E-/P-selectin expression in tissue microvasculature drives myeloid cell recruitment crucial for the development of the disease. Increased E-selectin expression and mononuclear phagocyte infiltration have been similarly observed in cases of transplantation rejection, including human renal ( 210 , 211 ), lung ( 212 – 214 ), and cardiac ( 215 – 217 ) rejection and in acute graft versus host disease ( 218 – 221 ).…”

Animal models for diabetes: Understanding the pathogenesis and finding new treatments

“…In the studies that use drugs to reverse diabetes, success is often only seen in the early stages of the disease with blood glucose concentrations only mildly hyperglycaemic [60][61][62][63]. Pancreas and islet transplantation is effective in reversing hyperglycaemia in BB rats and NOD mice if immune suppression or costimulatory blockade is administered [64][65][66][67] or if an encapsulation device is used [68].…”