Clinical impact of real-time evaluation of the biological activity and degradation of hepatocyte growth factor

Abstract

Hepatocyte growth factor (HGF) is essential for injury repair. Despite high HGF levels in chronic ulcers, up-regulation of HGF receptor in ulcer tissue and decreased biological activity of HGF in ulcer secretions have been observed. With a surface plasmon resonance-based method, we assessed the binding of HGF to antibodies, receptors, and the basement membrane and identified binding interactions that are indispensable for the biological activity of HGF. Recombinant HGF (rHGF) lots were tested for activity, structural integrity, and degradation, and the results were verified in an in vitro model of cell injury. Biologically active rHGF, as well as plasma from healthy volunteers, bound to heparan sulphate proteoglycan (HSPG) and to anti-HGF antibodies. Decreased binding to HSPG was the first event in rHGF degradation. This study established the feasibility of identifying patients with chronic inflammation who need exogenous HGF and of using ligand-binding assessment to evaluate rHGF lots for biological activity.

Differences in the expression of hepatocyte growth factor in acute and chronic bowel inflammation—Implications for diagnosis?

“…In recent years, however, several groups have focused on assessing HGF during inflammation and cancer [10]. We previously showed that patients with chronic/acute skin ulcers [17,18], chronic renal failure [31], ischemic heart disease [32], and periodontitis [33] express a high concentration of HGF but a decreased binding affinity of HGF to glucosaminoglycans. In the present study, we observed a higher concentration of HGF in both acute inflammation (i.e., C. difficile infection) and chronic inflammation (i.e., ulcerative and microscopic colitis and cancer) compared to healthy controls.…”

“…Despite being present in high concentrations, HGF may appear in a form with reduced biological activity, characterised by reduced binding affinity to HSPG and dextran sulphate and decreased regeneration in a model of cell injury (i.e., mouse skin epithelial cells, CCL-53.1) [27,28]. HSPG and dextran sulphate are both glucosaminoglycans and bind to the same epitope on the HGF molecule [18].…”

“…In SPR systems, biologically active HGF exhibits binding affinity for HSPG [17], which is decreased after the addition of sulphated oligosaccharides, such as the glycosaminoglycan/dextran sulphate [18]. In addition, biosensors coated with dextran sulphate allow the detection of HGF in cellular medium [19].…”

“…A potential therapeutic strategy with exogenous HGF in the treatment of chronic IBD has been proposed [20,21]. Taking advantage of previous studies and observations [17,18], we investigated and compared the presence, amount, and binding affinity of HGF in faecal samples from patients with acute and chronic bowel diseases.…”

A methachromatic-based experimental model for identification of bowel as the focus of an acute inflammation

“…Biosensor technology, such as surface plasmon resonance (SPR), is a reproducible method for real-time evaluation of binding affinity to ligands and receptors [5]. Biologically active HGF shows binding affinity to HSPG in SPR system and the affinity is decreased after addition of sulphated oligosaccharides such as DS to the samples [6,7]. Biosensor coated with DS was recently introduced for detection of HGF in cellular medium [8].…”