The association between skin collagen glucosepane and past progression of microvascular and neuropathic complications in type 1 diabetes

doi:10.1016/j.jdiacomp.2012.10.004

Journal of Diabetes and its Complications

Volume 27, Issue 2, March–April 2013, Pages 141-149

https://doi.org/10.1016/j.jdiacomp.2012.10.004 Get rights and content

Abstract

Purpose

We determined the association between novel and acid-labile skin collagen-linked advanced glycation endproducts (AGEs) and the progression of microvascular and neuropathic complications from baseline to near study closeout in the Diabetes Control and Complications Trial (DCCT).

Methods

From a skin biopsy obtained near the close of the DCCT, proteolytic collagen digests were analyzed by liquid chromatography/mass spectrometry (LC/MS/MS) for glucosepane (GSPNE), glyoxal and methylglyoxal hydroimidazolones (G-H1 and MG-H1) and the glycation product fructose-lysine (FL) using isotope dilution method.

Results

GSPNE and MG-H1 correlated with age and diabetes duration (P < 0.02), while GSPNE and FL correlated with the history of glycemia expressed as mean A1c (P ≤ 0.003). Age and duration-adjusted GSPNE and FL levels were lower in intensive (INT) vs. conventional (CONV) treatment subjects in the primary prevention DCCT cohort (P < 0.0001), and FL was lower in INT in the secondary intervention cohort (P < 0.0001). GSPNE was associated with increased incidence of retinopathy progression (odds ratio (OR) / unit increase in GSPNE: 2.5 for 3 step progression on the ETDRS scale, P = 0.003) and sustained ≥ 3 microaneurysms (MA) (OR = 4.8, P < 0.0001) from DCCT baseline up to the time of the biopsy, and prevalence of microalbuminuria or AER > 40 mg/24 h (OR = 5.3, P < 0.0001), and confirmed clinical neuropathy (OR = 3.4, P = 0.015) at the time of the biopsy. GSPNE adjusted for mean A1c remained significant for ≥ 3 MA (P = 0.0252) and AER (P = 0.0006). The strong association of complications with A1c was reduced or eliminated when adjusted for GSPNE.

Conclusions

Glucosepane is a novel AGE marker of diabetic complications that is robustly associated with nephropathic, retinopathic and neuropathic outcomes despite adjustment for A1c, suggesting that it could be one mediator of these complications with possible diagnostic implications.

Introduction

The discovery of HbA1c as a marker of cumulative glycemia over 2–3 months has greatly facilitated the design of clinical trials to probe the role of glycemia in the pathogenesis of diabetic complications. Notably, early and intensive treatment of glycemia over several years was associated with a lower progression rate of microvascular disease than conventional treatment in both type 1 and type 2 diabetes, e.g. in the Diabetes Control and Complications Trial (DCCT) and the United Kingdom Prospective Diabetes Study (UKPDS), respectively (The DCCT Research Group, 1993, UKPDS 33, 1998). The observed beneficial effects were sustained in observational follow-up studies of these cohorts, despite loss of the prior separation of hemoglobin A1c (A1c) between the treatment groups (White et al., 2010, Writing Team for the DDCT-EDIC, 2003).

While HbA1c (A1c) is a powerful measure of hyperglycemia and marker of complication risk (Selvin et al., 2011, Waden et al., 2009), several studies have examined the more stable advanced glycation endproducts (AGE's) both as markers of and as contributors to the pathogenesis of diabetic complications (for a review see (Monnier, Sell, & Genuth, 2005)). Major findings previously published revealed that skin collagen-linked AGE's are associated with past cumulative glycemia (A1c level), severity of diabetic microvascular complications and lower tissue levels of these collagen AGEs after intensive glycemic control in the DCCT (Monnier, Bautista, Kenny, et al., 1999). The AGE's, especially fructose-lysine (FL) (measured as furosine) and carboxymethyl-lysine (CML), were also predictors of the future risk of progression of retinopathy and nephropathy in EDIC (Genuth, Sun, Cleary, et al., 2005). Similar observations were made by other groups, especially with measurement of skin autofluorescence, certain skin collagen-linked oxidation markers and various serum carbonyl and oxidant stress markers associated with diabetic complications (Gerrits et al., 2008, Monnier et al., 2005, Yu et al., 2006).

The initial results we reported were based on 4 mm skin biopsies from 216 DCCT participants obtained near closeout of DCCT (1993) (Monnier et al., 1999). As only a limited number of chemically characterized advanced glycation endproducts were known, i.e. CML and pentosidine, two 4 mm skin samples were obtained with the second stored at − 80 °C under argon for future assay of novel AGEs. Data reported herein were obtained from using the stored, frozen biopsy obtained in 1993.

We determined the associations of the severity of DCCT microvascular complications and other DCCT clinical parameters with glucosepane (GSPNE), the single major AGE and protein crosslink discovered to date in human skin collagen (Sell et al., 2005), as well as carboxyethyl-lysine (CEL), and the hydroimidazolones of glyoxal (G-H1) and methylglyoxal (MG-H1). Although we previously reported furosine (Monnier et al., 1999), which is the acid hydrolyzed product of FL, the latter is still included in the AGE panel in this paper as a way to validate the reproducibility of the enzymatic (new assay) and acid hydrolysis methods (old assay), as well as the stability of FL after 16 years of storage. In the present study we aimed to investigate whether any of the newer collagen variables are 1) associated with cumulative glycemia, 2) reduced by intensive glycemia control, 3) associated with past progression and cross-sectional severity of complications, and if so, whether this association is independent of the cumulative A1c levels during the DCCT.

Section snippets

Research subjects

Two 4 mm skin punch biopsies were obtained 1–2 years prior to DCCT closeout in 1993 from 216 DCCT volunteers, 123 from the primary cohort [no retinopathy and AER < 40 mg/24 h at baseline] and 93 from the secondary cohort [mild to moderate retinopathy and AER < 200 mg/24 h at baseline]. The biopsies were stored under argon at − 80 °C. Retinopathy, AER, confirmed clinical neuropathy and A1c were assessed as previously described (The DCCT Research Group, 1993).

Chemicals

The source and preparation of isotopically

Participant characteristics

Table 1 recapitulates the clinical characteristics of the 216 participants at DCCT baseline and at closeout in the primary and secondary cohorts. As previously reported, the biopsy subgroups slightly differed from the non-participating DCCT cohorts (n = 1225) in that they were older (29 vs. 27 years), had slightly higher blood pressure (116 vs. 113 mm Hg), and had a higher rate of neuropathy at baseline (12% vs. 6%) (Monnier et al., 1999). At DCCT baseline subjects in the two treatment groups did

Discussion

Overwhelming evidence implicates chronic hyperglycemia as a major risk factor for the progression of microvascular and macrovascular disease in both type 1 and type 2 diabetes. A1c, as a measure of chronic hyperglycemia, emerged as a powerful glycemic marker of progression of micro- and macrovascular disease in multiple studies (Davis, 1992, Genuth, 1995, Janka et al., 1989, Stratton et al., 2000, The DCCT Research Group, 1995). The fact that complications still progress in some individuals in

Acknowledgments

A complete list of the members of the DCCT/EDIC Research Group is provided in the supplementary appendix published in New England Journal of Medicine, 2011; 365:2366–76. A list of the participating radiologists and technologists is shown in the online supplemental material published in Circulation, 2011: 124(16):1737–1746. Industry contributors have had no role in the conduct of EDIC, but have offered free or discounted supplies or equipment as a thank you to participants: Abbott, Animas,

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^☆: Funding: We thank the Juvenile Diabetes Foundation International (grant# 17-2010-318 to V.M.M.) and the NIDDK (DK-79432 to D.R.S.) for support. The DCCT/EDIC project is supported by contracts with the Division of Diabetes, Endocrinology and Metabolic Diseases of the National Institute of Diabetes and Digestive and Kidney Diseases, National Eye Institute, National Institute of Neurological Disorders and Stroke, the General Clinical Research Centers Program and the Clinical and Translation Science Centers Program, National Center for Research Resources, and by Genentech through a Cooperative Research and Development Agreement with the National Institute of Diabetes and Digestive and Kidney Diseases.

^☆☆: Duality of Interest: All authors have completed the Unified Competing Interest form at www.icmje.org/coi_disclosure.pdf (available on request from the corresponding author). V. Monnier is a paid consultant for L'Oreal. All other authors declare no competing interest.

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The association between skin collagen glucosepane and past progression of microvascular and neuropathic complications in type 1 diabetes☆,☆☆

Abstract

Purpose

Methods

Results

Conclusions

Introduction

Section snippets

Research subjects

Chemicals

Participant characteristics

Discussion

Acknowledgments

Free Radical Biology & Medicine

Kidney International

Biochimica et Biophysica Acta

Bioorganic & Medicinal Chemistry

The Journal of Biological Chemistry

The Journal of Biological Chemistry

Formation of immunochemical advanced glycosylation end products precedes and correlates with early manifestations of renal and retinal disease in diabetes

Diabetes

Controlling the false discovery rate: A practical and powerful approach to multiple testing

Journal of the Royal Statistical Society, Series B

Advanced glycation of fibronectin impairs vascular repair by endothelial progenitor cells: Implications for vasodegeneration in diabetic retinopathy

Investigative Ophthalmology & Visual Science

Diabetic retinopathy. A clinical overview

Diabetes Care

Increased dicarbonyl metabolism in endothelial cells in hyperglycemia induces anoikis and impairs angiogenesis by RGD and GFOGER motif modification

Diabetes

Regression and ANOVA with zero–one data: Measures of residual variation

Journal of the American Statistical Association

Mechanism of lysine oxidation in human lens crystallins during aging and in diabetes

The Journal of Biological Chemistry

The case for blood glucose control

Advances in Internal Medicine

Glycation and carboxymethyllysine levels in skin collagen predict the risk of future 10-year progression of diabetic retinopathy and nephropathy in the diabetes control and complications trial and epidemiology of diabetes interventions and complications participants with type 1 diabetes

Diabetes

Skin autofluorescence: A tool to identify type 2 diabetic patients at risk for developing microvascular complications

Diabetes Care

Risk factors for progression of background retinopathy in long-standing IDDM

Diabetes

Role of lipids in chemical modification of proteins and development of complications in diabetes

Biochemical Society Transactions

Serum levels of advanced glycation end products are increased in patients with type 2 diabetes and coronary heart disease

Diabetes Care

Muscle capillary basement membrane changes related to aging and to diabetes mellitus

Diabetes