Diabetic endothelial colony forming cells have the potential for restoration with glycomimetics

Alexander W W Langford-Smith,Alan M Jones,S Tawqeer Rashid,Ria Weston,Andrew J M Boulton,M Yvonne Alexander,Fiona L Wilkinson,Nicola Edwards,Frank L Bowling,Ahmad Hasan

doi:10.1038/s41598-019-38921-z

Abstract

Endothelial colony forming progenitor cell (ECFC) function is compromised in diabetes, leading to poor vascular endothelial repair, which contributes to impaired diabetic foot ulcer healing. We have generated novel glycomimetic drugs with protective effects against endothelial dysfunction. We investigated the effect of glycomimetic C3 on the functional capacity of diabetic ECFCs. ECFCs were isolated from healthy controls and patients with diabetes with neuroischaemic (NI) or neuropathic (NP) foot ulcers. Functionally, diabetic ECFCs demonstrated delayed colony formation (p < 0.02), differential proliferative capacity (p < 0.001) and reduced NO bioavailability (NI ECFCs; p < 0.05). Chemokinetic migration and angiogenesis were also reduced in diabetic ECFCs (p < 0.01 and p < 0.001), and defects in wound closure and tube formation were apparent in NP ECFCs (p < 0.01). Differential patterns in mitochondrial activity were pronounced, with raised activity in NI and depressed activity in NP cells (p < 0.05). The application of glycomimetic improved scratch wound closure in vitro in patient ECFCs (p < 0.01), most significantly in NI cells (p < 0.001), where tube formation (p < 0.05) was also improved. We demonstrate restoration of the deficits in NI cells but not NP cells, using a novel glycomimetic agent, which may be advantageous for therapeutic cell transplantation or as a localised treatment for NI but not NP patients.

Highlights

Diabetic foot ulceration is a chronic complication in diabetes where tissue damage occurs due to neuropathy, ischemia and/or infection[1] and given its resistance to treatment, provides the impetus for development of novel healing modalities
Routine clinical assessments were performed for participants with diabetes and both NP and NI groups exhibited above normal range levels of HbA1c, LDL and triglycerides, with no significant difference detected between the groups (Additional File Table 1)
Endothelial colony forming progenitor cell (ECFC) were isolated from NI and NP patients and controls followed by characterisation by positive staining for endothelial cell markers, lack of a hematopoietic marker (CD45), the ability to incorporate acetylated-LDL and binding of the lectin UEA-114

Summary

Introduction

Diabetic foot ulceration is a chronic complication in diabetes where tissue damage occurs due to neuropathy, ischemia and/or infection[1] and given its resistance to treatment, provides the impetus for development of novel healing modalities. We use ECFCs12,22 to identify distinct functional differences between ECFCs isolated from human peripheral blood from NI vs NP diabetic patients to (i) determine whether they exhibit similar functional repair defects based on their in vivo environment and (ii) establish whether glycomimetic C3 enhances these functional deficits in vitro. These findings may allow us to stratify future treatment approaches for this patient group based on their clinical phenotype; NI or NP

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Conclusion