of the ER22/23EK Polymorphism in the Glucocorticoid Receptor Gene with Survival and C-Reactive Protein Levels in Elderly Men

Elisabeth F. C. van Rossum, MD, Richard A. Feelders, MD, PhD, Annewieke W. van den Beld, MD, PhD, Andre´ G. Uitterlinden, PhD, Joop A. M. J. L. Janssen, MD, PhD, Wietske Ester, MD, Albert O. Brinkmann, Diederick E. Grobbee, PhD, Frank H. de Jong, PhD, Huibert A. P. Pols, MD, Jan W. Koper, PhD, Steven W. J. Lamberts, MD, PhD

Interleukin 6 was determined by a commercially available im- mulite assay.

PURPOSE: We recently demonstrated that a polymorphism in codons 22 and 23 of the glucocorticoid receptor gene is associ- ated with relative glucocorticoid resistance, greater insulin sen- sitivity, and lower total and low-density lipoprotein cholesterol levels. In the present study, we investigated whether the ER22/ 23EK polymorphism is associated with survival, cholesterol lev- els, and two predictors of mortality: serum C-reactive protein and interleukin 6 levels.   METHODS: We studied 402 men (mean [ SD] age, 77.8

RESULTS: After a follow-up of 4 years, 73 (19%) of 381 non- carriers died, while none of the 21 ER22/23EK carriers had died  (P 0.03). C-reactive protein levels were about 50% lower in ree= ER22/23EK carriers (P 0.01). There were no differences in interleukin 6 levels.

CONCLUSION: Carriers of the ER22/23EK polymorphism havebettersurvivalthannoncarriers,aswellaslowerC-reactive protein levels. Am J Med. 2004;117:158–162. ©2004 by Elsevier Inc.

METHODS: We studied 402 men (mean [ SD] age, 77.8 method using a latex-enhanced immunoephelometric

the liver, and is elevated in patients with unstable angina or heart failure (8–11). Therefore, we investigated whether C-reactive protein and interleukin 6 levels, as well as survival, were associated with the ER22/23EK polymorphism of the glucocorticoid receptor gene.

in codons 22 and 23 of the glucocorticoid receptor gene (GAGAGG [GluArg, or ER] 3 GAAAAG [GluLys, or EK]) is associated with relative glucocorticoid resistance (2). This ER22/23EK variant was also associated with greater insulin sensitivity and lower total and low-density lipoprotein (LDL) cholesterol levels. Furthermore, we found that the number of ER22/23EK carriers was signif- icantly higher in the older half of the sample, suggesting that the polymorphism had a beneficial effect on survival.

METHODS

Subjects

We recruited 402 men, aged 73 years or older, by a letter that was sent to the oldest men in Zoetermeer, The Neth- erlands. Subjects were eligible to participate if they were physically and mentally able to visit the study center in- dependently. No additional health-related criteria were used. Medications taken for more than 6 months were recorded. Data on vital status of the participants and causes of death during 4-year follow-up were obtained by contacting the participants’ general practitioners. Before the start of the study, which received the approval of the Medical Ethics Committee of the Erasmus Medical Cen- ter, all subjects had given their written informed consent to participate.

The ER22/23EK variant might also affect the inflam- matory response, and elevated levels of two inflammatory markers—C-reactive protein and interleukin 6—are as- sociated with mortality in the elderly (3). C-reactive pro- tein is an independent risk factor for cardiovascular events (4–7). Interleukin 6 stimulates the synthesis of C- reactive protein, as well as other acute phase proteins in

From the Departments of Internal Medicine (EvR, RF, AvdB, AU, JJ, WE, FdJ, HP, JK, SL) and Reproduction and Development (AB), Eras- mus Medical Center, Rotterdam, The Netherlands; and Julius Center (DG), University of Utrecht, The Netherlands.

Measurements

This research project was supported by a grant from NWO and Re- search Institute of Diseases in the Elderly.

Weight and height were measured, and the body mass index (kg/m2) was calculated. Blood pressure was mea- sured in sitting position at the right upper arm with a random-zero sphygmomanometer. Total fat mass, trunk fat mass, and lean body mass were measured using dual- energy X-ray absorptiometry (Lunar Corp., Madison,

Manuscript submitted June 17, 2003, and accepted in revised form January 29, 2004.

Wisconsin) (12). Quality assurance, including calibra- tion, was performed every morning, using the standard provided by the manufacturer.

were not normally distributed, logarithmic transforma- tions were applied to normalize them or nonparametric tests (Mann-Whitney U test) were used. Continuous - *éSpS variables are reported as mean SD or median with the interquartile range. Survival was analyzed using the Kaplan-Meier procedure and log-rank test. To study the association between C-reactive protein levels and mortal- ity, C-reactive protein levels were divided in two groups (high vs. low, based on the median value). Cox propor- tional hazards models were used to analyze this relation, adjusting for genotype, diabetes, and health status. Cor- relations between C-reactive protein levels and parame- ters of body composition and cortisol levels were calcu- <0 lated using Spearman’s correlation. P 0.05 was considered statistically significant.