Originl Article Yonsei Med J 2015 Nov;56(6):1604-1612 pissn: 0513-5796 eissn: 1976-2437 The Impct of CDH13 Polymorphism nd Sttin Administrtion on TG/HDL Rtio in Crdiovsculr Ptients Jung Rn Choi 1, Yngsoo Jng 2, Sungjoo Kim Yoon 3, Jong Keun Prk 3, Sungin Richrd Sorn 4, Mi-Young Prk 5, nd Myoungsook Lee 1 1 Deprtment of Food nd Nutrition, nd Reserch Institute of Oesity Sciences, Sungshin Women s University, Seoul; 2 Crdiovsculr Genome Center, Crdiology Division, Deprtment of Internl Medicine, Severnce Crdiovsculr Hospitl, Yonsei University College of Medicine, Seoul; 3 Reserch Institute of Moleculr Genetics, The Ctholic University of Kore, Seoul; 4 School of Medicine, The Ctholic University of Kore, Seoul; 5 Institute of Helth nd Environment, Grdute School of Pulic Helth, Seoul Ntionl University, Seoul, Kore. Purpose: is expressed in dipose tissue, nd is ffected y smoking, oesity, nd genetic fctors, such s CDH13 polymorphism, contriuting to the development of coronry vsculr diseses (CVDs). Mterils nd Methods: We investigted the effect of genetic vritions of CDH13 (rs3865188) on lood chemistry nd diponectin levels in 345 CVD ptients undergoing sttin-free or sttin tretment. Results: Genetic vrition in CDH13 ws significntly correlted with severl clinicl fctors, including diponectin, distolic lood pressure, triglyceride (TG), nd insulin levels. Sujects with the T llele (mutnt form) hd significntly lower diponectin levels thn those with the A llele. Totl cholesterol (TC), low-density lipoprotein cholesterol (LDLc), TG/high-density lipoprotein cholesterol (HDLc) rtio, nd HDL3 sutype were mrkedly decresed in sttin treted sujects regrdless of hving the A or T llele. TG nd TG/HDL in the sttin-free group with genotype of the rs3865188 ws higher thn in the others ut they were not different in the sttin-treted sujects. We oserved significnt difference in diponectin levels etween ptients with the A nd T lleles in the sttin-free group; menwhile, no difference in diponectin levels ws noted in the sttin group. Plsm levels of other cytokines, leptin, visftin, interleukin-6 (IL-6), nd tumor necrosis fctor-α (TNF-α), were not different mong the CDH13 genotypes ccording to sttin dministrtion. Body mss index (BMI), TG, insulin, HDL3, nd TG/HDL rtio showed negtive correltions with diponectin levels. Conclusion: Plsm diponectin levels nd TG/HDL rtio were significntly different ccording to vrints of CDH13 nd sttin dministrtion in Koren ptients with CVD. Key Words:, CDH13, sttin, TG/HDL, HDL3 Received: Ferury 2, 2015 Revised: July 21, 2015 Accepted: July 21, 2015 Co-corresponding uthors: Myoungsook Lee, PhD, Deprtment of Food nd Nutrition, nd Reserch Institute of Oesity Sciences, Sungshin Women s University, 55 Doong-ro 76g-gil, Gnguk-gu, Seoul 01133, Kore. Tel: 82-2-920-7211, Fx: 82-2-920-2078, E-mil: mlee@sungshin.c.kr nd Yngsoo Jng, MD, PhD, Crdiovsculr Genome Center, Crdiology Division, Deprtment of Internl Medicine, Severnce Crdiovsculr Hospitl, Yonsei University College of Medicine, 50-1 Yonsei-ro, Seodemun-gu, Seoul 03722, Kore. Tel: 82-2-2228-8445, Fx: 82-2-393-2041, E-mil: jngys1212@yuhs.c The uthors hve no finncil conflicts of interest. Copyright: Yonsei University College of Medicine 2015 This is n Open Access rticle distriuted under the terms of the Cretive Commons Attriution Non-Commercil License (http://cretivecommons.org/ licenses/ y-nc/3.0) which permits unrestricted non-commercil use, distriution, nd reproduction in ny medium, provided the originl work is properly cited. INTRODUCTION Some dipokines re produced in dipose tissue, while others re prtilly secreted y infiltrting mcrophges nd stroml cells. 1 Among dipokines, diponectin is only expressed y dipocytes, nd stroml vsculr cells of dipose tissue. is known to increse insulin-stimulted glucose uptke y enhncing Akt phosphoryltion. Plsm diponectin levels hve een found to e correlted with ody mss index (BMI), type 2 dietes mellitus (T2DM), nd coronry vsculr disese (CVD). Additionlly, decresed expression, secretion, nd circulting levels of diponectin hve een reported in oese popultions. 1-3 tretment reduces the expression of endotoxin-induced pro-inflmmtory cytokines nd 1604 www.eymj.org
Jung Rn Choi, et l. increses nti-inflmmtory interleukin-10 (IL-10) expression in monocytes nd mcrophges. Exogenous dministrtion of diponectin lso decreses llergic irwy inflmmtion in mice. 4 In spite of its nti-inflmmtory nd nti-therogenic effects, diponectin is potentilly ssocited with dverse clinicl outcomes, such s mortlity. As plsm diponectin levels re strongly influenced y single nucleotide polymorphisms (SNPs) of the gene encoding T-cdherin [the cdherin 13 preprotein (CDH13)], severl studies hd een conducted to exmine the possile ssocitions etween CDH13 genotype nd plsm diponectin levels. 5 CDH13, 95 kd glycoprotein, is n typicl memer of the cdherin fmily of cell dhesion molecules. 4 Unlike clssicl cdherins, it hs low dhesive chrcters nd is ttched to the cell memrne vi glycosylphosphtidylinositol nchor, s it lcks oth trnsmemrne nd cytoplsmic domins. 6 CDH13 functions s negtive regulting molecule during the development of the nervous system nd is involved in migrtory processes, tumorigenesis, nd ngiogenesis. 7,8 Moreover, CDH13 is involved in endoplsmic reticulum (ER) stress responses. Severl studies suggested CDH13 is upregulted in endothelil cells fter induction of ER stress nd protect these cells from poptosis in rection to the propoptotic response. 9 Another report oserved tht GRP78, moleculr chperone similr to CDH13, ws involved in pro-survivl responses to ER stress, s signling prtner to CDH13, long to the surfce of endothelil cells. 10 Recently, severl CDH13 SNPs, such s rs3865188, hve een reported to e determinnts of lood diponectin levels in Asins. The ssocition of rs11646213 in CDH13 nd lood pressure phenotype hs lso een investigted in Europen cohorts. Another SNP, rs4783244, ws lso shown to e ssocited with metolic syndrome phenotype in Tiwnese. 11 However, the ssocition etween CDH13 vritions nd crdiometolic disese is not completely cler. Sttins [or 3-hydroxy-3-methyl-glutryl (HMG)-CoA reductse inhiitors] re known to e effective in reducing risk of crdiovsculr events in ptients with CVD y lowering cholesterol synthesis. 12 Severl reports hve demonstrted tht sttin shows pleiotropic effects in modulting endothelil function, stle ngin pectoris, inflmmtion, nd thromosis. 13,14 However, only the impct of APOE on sttin tretment efficcy hs een reported, lthough the APOE4 llele is ssocited with poor response to sttin tretment. 15 Therefore, in this study, we imed to exmin the effect of CDH13 rs3865188 on diponectin levels nd lipid profiles in 345 sttin-free or sttintreted outptients enrolled with the Crdiovsculr Center t Severnce Hospitl in Seoul, Kore. MERIALS AND METHODS Study popultion A totl of 345 outptients were recruited from the Crdiovsculr Genome Center Registry from the Yonsei University College of Medicine, supported y The Ministry of Helth nd Welfre. Briefly, consecutive sujects who visited the Crdiovsculr Center t Severnce Hospitl in Seoul, Kore from July 2008 to Decemer 2009 were prospectively screened for dyslipidemi [triglyceride (TG) >150 mg/dl; high-density lipoprotein-cholesterol (HDLc) <40 mg/dl for men, HDL <50 mg/ dl for women]. Sujects who exhiited positive results for dyslipidemi on three seprte visits nd ptients who hd tken nti-hyperlipidemic medictions such s sttins were lso included. For further nlysis of the ssocition etween genetic vrints nd other metolic trits, the study popultion ws divided into two groups: sttin-free group nd sttin group. We nlyzed the effects of CDH13 rs3865188 on lood chemistry with sttin groups compred to the sttin free groups. This study ws conducted ccording to the guidelines of the Declrtion of Helsinki, nd ws pproved y the Institutionl Review Bord of Yonsei University Medicl Center (IRB#: 4-2001- 0039). Written informed consent ws otined from ll study prticipnts. Clinicl dt nd lood smples On the dy of enrollment, clinicl dt including demogrphic vriles nd medicl history were recorded. We clculted BMI s rtio of weight (kg) divided y the height (m 2 ). Blood lipid profiles [totl cholesterol (TC), low-density lipoproteincholesterol (LDLc), HDLc, TG, nd HDL sutypes], fsting lood sugr (FBS), insulin, enzymes relted to HDL metolism [lecithin cholesterol cyltrnsferse (LC) nd cholesteryl ester trnsfer protein (CETP)], nd dipocytokines [diponectin, leptin, monocyte chemottrctnt protein-1 (MCP-1), visftin, IL-6, tumor necrosis fctor-α (TNF-α)] were otined from serum or plsm collected fter n overnight fst nd kept t -80 C until further nlysis. FBS, TC, nd TG were mesured using Hitchi-7600 nlyzer (Hitchi Ltd., Tokyo, Jpn), nd LDLc ws clculted y Friedewld eqution. The 600 µl of fresh serum, kept t 4 C for less thn 1 dy, used for HDL suclsses fter HDL frction ws isolted y new micro-ultrcentrifugtion t 1.063<d<1.21 g/ml (Hitchi CS150GXL, CS140 fixed ngle rotor, Tokyo, Jpn), nd seprtion of HDL sufrctions were conducted s previously reported. 16 All cytokines, s well s LC nd CETP, were mesured y enzyme immunossy using n ELISA kit (Diichi Pure Chemicls, Tokyo, Jpn). Serum diponectin nd insulin levels were mesured y n enzyme-linked immunosorent ssy (Mesdi Co. Ltd., Seoul, Kore). 17 The intr- nd inter-ssy vrinces for diponectin were 6.3 7.4% nd 4.5 8.6%, respectively. Sequencing nd genotyping PCR primers were designed to independently mplify CDH13 frgments. Primer sequences re ville on request. PCR products were purified nd then sequenced using BigDye Termintor Cycle Sequencing Kit (version 3.1, ABI, Foster City, 1605
CDH13 Gene nd Sttin on TG/HDL in CVD CA, USA) nd n ABI 3730 1 utomted sequencer (Applied Biosystems, Foster City, CA, USA). The sequencing primers were the sme s those used for PCR mplifiction. SNPs identified in the CDH13 gene y whole gene sequencing were genotyped. Genomic DNA ws extrcted from 5 ml of peripherl venous lood using commercilly ville isoltion kit (QuickGene SP Kit DNA whole lood, Fujifilm, Tokyo, Jpn). Genotyping ws performed using the TqMn fluorogenic 5 nuclese ssy (ABI). Sttisticl nlysis Group differences for ctegoricl vriles were ssessed y chi-squre test, nd continuous vriles were exmined y Student s t-test. The ssocition etween genotype nd diponectin level ws evluted using odds rtios (ORs) nd 95% confidence intervls (CIs) from chi-squre tests nd logistic regression nlyses. The results re presented s men±stndrd error rnge. p-vlues for ech genotype nd llele were evluted y χ 2 nlysis nd Fisher s exct test. A p-vlue with three deciml plces ws tken to indicte sttisticlly significnt differences etween sttin-free nd sttin-treted sujects of the study. Associtions etween clinicl vriles, such s ge, BMI, nd TC, were nlyzed y Person correltion test. All nlyses were performed using SPSS softwre, version 18.0 (SPSS Inc., Chicgo, IL, USA). RESULTS Chrcteristics of the study popultion ccording to genotype nd lleles of rs3865188 Chrcteristics of the popultion re shown in Tle 1. The 345 Tle 1. Anthropometric nd Clinicl Chrcteristics of Crdiovsculr Disese Ptients According to Allele nd Genotype of CDH13 Gene SNP (rs3865188) Vriles Totl Genotype Allele (n=345) (n=170) (n=128) (n=36) p vlue* A (n=469) T (n=201) p vlue* 5.81±3.70 006.25±3.76 005.79±3.66 003.97±3.29 0.046 6.13±3.72 5.08±3.60 0.016 Age (yr) 62.56±1 63.31±10.07 61.91±9.53 62.92±10.38 0.481 62.93±9.92 62.28±9.80 0.451 BMI (kg/m 2 ) 25.09±3.25 24.91±3.23 25.40±3.49 24.78±2.44 0.371 25.04±3.31 25.17±3.15 0.618 SBP (mm Hg) 116.73±13.49 116.90±12.27 115.29±14.15 118.33±12.98 0.382 116.47±12.79 116.40±13.75 0.929 DBP (mm Hg) 71.39±8.30 72.09±7.70 69.96±7.81 72.22±7.96 0.052 71.52±7.77 70.79±7.90 0.717 TC (mg/dl) 150.50±38.74 147.64±37.80 155.60±39.89 146.88±36.84 0.176 149.82±38.47 152.46±38.87 0.419 TG (mg/dl) 142.71±81.29 134.96±78.14 147.77±80.39 158.13±90.92 0.184 138.46±78.80 151.50±84.00 0.055 HDLc (mg/dl) 39.96±9.69 40.47±9.66 39.49±9.41 39.63±10.38 0.668 40.20±9.58 39.54±9.72 0.411 LDLc (mg/dl) 85.72±31.51 83.04±30.57 90.88±32.31 81.30±31.18 0.067 85.18±31.19 87.43±32.09 0.403 TG/HDL 3.94±2.97 3.67±2.84 4.10±2.96 4.48±3.39 0.226 3.79±2.87 4.24±3.11 0.070 Insulin 10.91±11.26 12.36±14.19 9.87±8.12 8.70±4.84 0.088 11.66±12.80 9.45±7.11 6 HOMA-IR 3.63±5.50 3.96±7.02 3.57±4.01 2.41±1.37 0.328 3.85±6.31 3.15±3.35 0.069 FBS 12±52.84 118.49±42.13 134.00±65.52 116.13±41.93 0.026 122.73±5 127.57±58.58 0.316 CRP 0.33±0.78 0.26±0.60 0.49±1.07 0.23±0.41 0.344 0.31±0.75 0.39±0.87 0.550 Leptin 29.55±30.05 27.25±29.86 35.36±34.14 22.61±12.87 0.145 29.28±31.05 30.36±28.38 0.751 MCP-1 44.81±38.40 40.08±34.28 53.97±44.53 42.03±36.00 0.084 43.55±37.44 49.29±41.46 0.219 Visftin 1.87±0.12 1.86±0.12 1.87±0.12 1.89±0.13 0.489 1.86±0.12 1.88±0.12 0.208 IL-6 37.48±56.05 40.40±58.46 38.56±60.67 24.70±26.81 0.530 39.94±58.78 33.12±50.40 0.312 TNF-α 26.95±32.07 29.68±39.62 23.30±19.95 22.92±15.81 0.406 28.09±35.71 23.15±18.29 0.087 BUN 16.38±4.77 16.33±4.55 16.13±4.54 17.63±6.39 0.344 16.28±4.53 16.65±5.26 0.414 Uric cid 5.37±1.45 5.31±1.44 5.37±1.50 5.59±1.31 0.699 5.33±1.45 5.45±1.43 0.418 HDL2 31.79±3.08 31.96±3.01 31.52±3.08 31.89±3.23 0.509 31.83±3.03 31.65±3.12 0.505 HDL2 22.27±1.57 22.19±1.72 22.39±1.39 22.15±1.58 0.550 22.24±1.63 22.30±1.45 0.679 HDL3 18.56±1.70 18.45±1.71 18.68±1.62 18.51±1.71 0.548 18.52±1.68 18.62±1.64 0.495 HDL3 12.73±1.51 12.70±1.47 12.81±1.55 12.63±1.57 0.787 12.73±1.49 12.75±1.55 0.919 HDL3c 14.55±2.33 14.54±2.46 14.58±2.20 14.71±2.45 0.932 14.55±2.38 14.63±2.28 0.719 LC mss 8.72±2.76 8.62±2.84 8.80±2.83 8.98±2.36 0.809 8.67±2.83 8.87±2.64 0.491 CETP mss 1.62±0.67 1.55±0.56 1.67±0.68 1.63±0.74 0.466 1.58±0.59 1.66±0.70 0.312 LC/CETP 6.13±2.80 6.15±2.78 6.06±2.76 6.47±3.18 0.821 6.12±2.77 6.21±2.91 0.782 SNP, single nucleotide polymorphism; BMI, ody mss index; SBP, systolic lood pressure; DBP, distolic lood pressure; TC, totl cholesterol; TG, triglyceride; HDLc, high-density lipoprotein cholesterol; LDLc, low-density lipoprotein cholesterol; FBS, fsting lood sugr; CRP, C-rective protein; MCP-1, monocyte chemottrctnt protein-1; IL-6, interleukin 6; TNF-α, tumor necrosis fctor-α; HOMA-IR, homeostsis model ssessment of insulin resistnce. Vlues with different lphets re significntly different mong the groups y ANOVA t p<5. *p vlue ws estimted y ANOVA nd t-test. 1606
Jung Rn Choi, et l. sujects recruited were ged 62.56±10.0 yers, nd 67.3% (n= 233) of the totl popultion were men. Sujects with the CDH13 SNP showed significnt differences in severl clinicl fctors, including diponectin level; distolic lood pressure (DBP); nd levels of FBS, insulin, nd TG (Tle 1). Sujects with the T llele (mutnt form) hd significntly lower diponectin nd insulin levels nd significntly higher plsm TG thn those with the A llele (wild-type form) (p<0.05). In contrst, lleles of the CDH13 gene showed divergent trend when compred with the genotype. There were significnt differences in FBS level nd DBP etween heterozygous sujects with the llele nd homozygous sujects with n or llele, which my hve contriuted to the non-significnt differences oserved in the FBS level nd DBP etween A nd T lleles. Other lood chemistry dt, such s lipid profiles with HDL sutypes or inflmmtory cytokines, did not differ y CDH13 genotype or llele types. CDH13 vrints nd sttin tretment on lipid risk fctors One SNP (rs3865188) ws significntly ssocited with diponectin level. In the dditive model, diponectin level ws not ssocited with decresed risk of dyslipidemi (OR=0.732, 95% CI, 0.349 1.545, p=0.406) (dt not shown). Sujects with the A llele hd higher diponectin levels thn those with the T llele in the sttin-free group, ut there were no differences in sttin groups (Tle 2). There ws lso difference in TG/ HDL levels etween sujects with the T llele nd those with the A llele in the sttin-free group. Moreover, there ws no difference in TG/HDL nd diponectin levels in the sttin group. The sujects with minor T llele of rs3865188 hd higher LDLc nd MCP-1 levels, wheres those with rs3865188 hd lower insulin levels in the sttin group (p=3). Sttin Tle 2. Anthropometric nd Clinicl Chrcteristics of Crdiovsculr Disese Ptients with CDH13 SNP (rs3865188) Alleles According to Sttin Administrtion Vriles Sttin-free group (n=177) Sttin group (n=168) Totl A T p vlue Totl A T p vlue p vlue* 5.83±3.45 6.25±3.70 4.78±2.56 7 5.79±4.00 5.99±3.77 5.45±4.61 0.441 0.073 Age (yr) 62.31±10.13 62.92±9.72 62.22±10.15 0.543 62.87±9.84 62.97±10.14 62.43±9.44 0.654 0.898 BMI (kg/m 2 ) 24.96±.3.07 24.77±3.10 25.21±2.91 0.228 25.23±3.41 25.31±3.48 25.15±3.40 0.709 0.341 SBP (mm Hg) 116.88±13.74 116.02±12.26 116.81±14.59 0.606 116.52±13.21 116.87±13.32 115.82±12.78 0.516 0.855 DBP (mm Hg) 71.78±8.26 71.61±7.33 71.32±7.23 0.733 74.66±48.40 74.09±41.42 76.79±64.49 0.654 0.586 TC (mg/dl) 161.62±38.52 161.57±38.12 161.81±38.06 0.958 138.77±35.47 137.86±35.05 141.92±37.22 0.346 <1 TG (mg/dl) 149.81±85.70 140.85±76.21 164.83±97.75 0.026 135.23±75.90 136.04±81.44 136.46±62.29 0.936 0.017 HDLc (mg/dl) 39.37±10.13 39.94±9.92 38.83±10.54 0.345 40.58±9.19 40.46±9.24 40.35±8.69 0.901 0.540 LDLc (mg/dl) 95.23±31.61 96.46±30.80 c 92.46±32.69 c 0.277 75.71±28.21 73.71±27.19 81.77±30.57 0.020 <1 TG/HDL 4.23±3.15 3.87±2.71 4.81±3.78 0.022 3.64±2.73 3.71±3.03 3.59±1.93 0.745 7 Insulin 10.78±11.69 11.24±13.22 9.86±8.10 0.328 11.06±10.82 12.09±12.36 8.99±5.80 3 0.114 HOMA-IR 3.80±6.84 4.03±8.00 3.32±3.72 0.392 3.44±3.54 3.66±3.80 2.96±2.88 0.076 0.418 FBS 125.16±54.81 123.63±53.75 127.33±58.63 0.569 124.84±50.86 121.81±45.98 127.84±58.84 0.388 0.730 CRP 0.31±0.57 0.29±0.55 0.38±0.62 0.435 0.36±1.03 0.35±0.98 0.41±1.20 0.845 0.905 Leptin 30.66±31.88 31.36±34.95 29.66±24.10 0.738 28.17±27.75 26.72±25.36 31.24±33.29 0.346 0.668 MCP-1 44.76±43.61 45.94±43.76 43.29±44.42 0.703 44.88±31.05 40.61±27.63 56.89±36.47 0.011 0.134 Visftin 1.87±0.11 1.88±0.12 1.87±0.10 0.940 1.86±0.13 1.85±0.12 1.89±0.15 0.067 0.148 IL-6 38.30±56.34 38.90±56.35 38.01±57.96 0.922 36.47±56.03 41.23±61.89 26.93±38.53 0.090 0.569 TNF-α 29.54±39.36 31.50±43.89 22.44±20.71 0.053 23.74±19.40 23.88±21.22 24.05±14.84 0.980 0.140 BUN 16.10±4.56 15.90±4.22 16.68±5.30 0.183 16.72±4.99 16.72±4.86 16.61±5.24 0.874 0.327 Uric cid 5.31±1.51 5.24±1.52 5.38±1.46 0.530 5.43±1.39 5.41±1.38 5.51±1.40 0.598 0.567 HDL2 31.50±2.94 32.06±3.25 32.03±3.12 0.393 32.11±3.21 32.06±3.25 32.03±3.12 0.944 0.173 HDL2 22.20±1.42 22.33±1.86 22.42±1.42 0.808 22.34±1.73 22.33±1.86 22.42±1.42 0.715 0.512 HDL3 18.59±1.51 18.52±1.93 18.61±1.79 0.550 18.52±1.89 18.52±1.93 18.61±1.79 0.694 0.925 HDL3 12.92±1.37 12.54±1.67 12.53±1.53 0.897 12.53±1.63 12.54±1.67 12.53±1.53 0.969 0.021 HDL3c 14.76±2.16 14.33±2.61 14.33±2.30 0.622 14.33±2.50 14.33±2.61 14.33±2.30 0.996 0.116 LC mss 8.71±2.78 8.66±2.83 8.96±2.73 0.464 8.73±2.74 8.68±2.84 8.77±2.57 0.819 0.890 CETP mss 1.64±0.75 1.62±0.62 1.62±0.76 0.990 1.59±0.58 1.55±0.57 1.69±0.63 0.111 0.488 LC/CETP 6.14±2.89 5.98±2.76 6.58±3.10 0.164 6.12±2.72 6.26±2.77 5.83±2.67 0.283 0.363 SNP, single nucleotide polymorphism; BMI, ody mss index; SBP, systolic lood pressure; DBP, distolic lood pressure; TC, totl cholesterol; TG, triglyceride; HDLc, high-density lipoprotein cholesterol; LDLc, low-density lipoprotein cholesterol; FBS, fsting lood sugr; CRP, C-rective protein; MCP-1, monocyte chemottrctnt protein-1; IL-6, interleukin 6; TNF-α, tumor necrosis fctor-α; HOMA-IR, homeostsis model ssessment of insulin resistnce. Vlues with different lphets re significntly different mong the groups y ANOVA t p<0.05. *p vlue ws estimted y ANOVA, p vlue ws estimted y t-test. 1607
CDH13 Gene nd Sttin on TG/HDL in CVD tretment strongly decresed TC, LDLc, TG/HDL rtio, nd HDL3 sufrctions regrdless of hving the A or T llele, respectively. In the sttin-free group, sujects with the genotype of the rs3865188 SNP hd lower diponectin levels (p=0.032) nd higher TG levels (p=0.025) (Fig. l). In the sttin group, however, the genotype ws strongly ssocited with reductions in TG levels nd in TG/HDL. Plsm diponectin levels were significntly incresed for individuls with the genotype nd those who underwent sttin tretment. Trends in inflmmtion mrkers in sujects who underwent sttin tretment Sujects in the sttin group with the genotype hd higher MCP-1 levels thn ptients in the sttin-free group with the sme genotype (57.92±40.90 vs. 26.14±22.39). Sttin tretment did not effect reductions in MCP-1 mong persons with the genotype (Fig. 1). In terms of llele distriution, there ws significnt difference in MCP-1 levels in sujects with the T llele, compred to those in sujects with the A llele, in the sttin group (p=9) (Tle 2). On the other hnd, individuls with the genotype in the sttin group hd higher leptin levels thn wild type ptients nd mutnt homozygous ptients. Plsm levels for other cytokines, leptin, visftin, IL-6, nd TNF-α did not differ ccording to CDH13 genotype or sttin dministrtion. TNF-α levels were lower for the T llele thn the A llele (Tle 2). Risk fctors relted to diponectin level Prmetric Person correltion nlysis of the dt reveled tht the diponectin level is significntly positively correlted with sex (r=0.339, p<1), HDL level (r=0.353, p<1), nd HDL2 level (r=0.367, p<1). Menwhile, TG level (r=-0.210, p=5), BMI (r=-0.224, p=3), insulin level (r=-0.252, p= 1), HDL3 (r=-0.325, p<1), nd TG/HDL rtio (r=-0.227, p=2) showed negtive correltions (dt not shown). In the sttin-free nd sttin groups, liner regression nlysis ws performed to estimte the reltive contriutions of clinicl vriles, such s BMI, TG level, insulin level, nd HDL2 frction, to the inter-individul vritions in diponectin level. Therein, TG level ws ssocited with diponectin level in sttin-free individuls with the T llele, not the A llele. In the sttin group, there ws no ssocition etween TG nd diponectin levels in individuls with the T llele (Fig. 2). Plsm diponectin level ws negtively ssocited with TG/HDL rtio in ptients with the genotype in the sttin-free group. However, the effect of the genotype ws not oserved upon sttin tretment. DISCUSSION This study is the first to suggest significnt ssocition etween SNPs locted within CDH13 nd sttin tretment on plsm diponectin levels nd lipid profiles in Koren crdiovsculr ptients with dyslipidemi. First, we found tht plsm diponectin levels decresed ccording to CDH13 genotypes, in order to from mjor to minor types. Although the mechnism of the trnscriptionl regultion of CDH13 is not cler, nucleotide vrint in the promoter region is sso- p=0.032 p=ns p=0.027 p=ns A 14 12 10 8 6 4 2 0 Sttin-free Sttin TG/HDL B 12 10 8 6 4 2 0 Sttin-free Sttin p=ns p=0.038 p=0.021 LDLc (mg/dl) 140 120 100 80 60 40 20 0 C Sttin-free Sttin D Sttin-free Sttin Fig. 1. Plsm levels of diponectin (A), TG/HDL (B), LDLc (C), HDL3 (D) sutypes ccording to CDH13 genotypes whether the stin ws tken or not. Vlues with different lphets re significntly different mong the CHD13 gene genotypes. TG, triglyceride; HDL, high-density lipoprotein; LDLc, low-density lipoprotein cholesterol. HDL3 15 14.5 14 13.5 13 12.5 12 11.5 11 1608
Jung Rn Choi, et l. Sttin-free group Sttin group 2 2 2 1 1 A 2 2 3 3 4 BMI B 2 2 3 3 BMI 2 2 2 1 1 C 10 20 30 40 50 TG D 20 40 60 TG 2 2 2 1 1 1 2 2 1 2 2 E TG/HDL rtio F TG/HDL rtio Fig. 2. Correltion of diponectin nd clinicl vriles ccording to genotype distriution of rs3865188. Body mss index (BMI) ws negtively correlted with diponectin in only the genotype (p=9) in contrst to the genotype in sttin group; menwhile, genotype cn e offset y genotype, compred to nd genotype (A nd B). Plsm diponectin ws relted with TG in A llele of ptients tking sttin (C nd D), nd it ws relted with TG/HDL rtio in the genotype in sttin-free group. However, with sttin tretment, the effect of genotype disppered (E nd F). TG, triglyceride; HDL, high-density lipoprotein. 1609
CDH13 Gene nd Sttin on TG/HDL in CVD cited with incresed promoter ctivity, nd lso plys crucil role in its expression. 18,19 Even though rs3865188 is not locted in the promoter region of CDH13, we demonstrted in the present study tht this vrint is positively ssocited with diponectin levels. 18 This is similr to the findings of previous studies, nd our results suggest tht the CDH13 vrint rs3865188 my regulte diponectin levels. The trditionl crdiovsculr risk fctors, including ge, sex, hypertension, smoking, hypercholesterolemi, nd ody mss index, hve een reported. 20 Another study investigted the role of T-cdherin expression in regulting diponectin levels, nd the involvement of CDH13 or diponectin levels in the development of crdiometolic diseses. 11 Furthermore, severl reports hve found tht CDH13 vrints hve modulting effects on metolic trits, nd they increse the risk of metolic syndrome, dietes mellitus, nd ischemic stroke. In contrst to these previous studies, rs3865188 vrints were not ssocited with risk of crdiometolic diseses including dyslipidemi in our study. However, TG, TG/HDL, nd diponectin levels ccording to CDH13 genotypes or llele types might e chnged y sttin dministrtion. This suggests tht increses in plsm diponectin levels upon sttin dministrtion re strongly ssocited with prticulr genes. We lso found tht CDH13 SNP (rs3865188) is strongly ssocited with metolic trits, including BP nd plsm levels of FBS, insulin, nd TG nd tht diponectin levels re not negtively ssocited with dyslipidemi descried y TG/HDL. Recently, n ssocition etween CDH13 SNP nd BP ws identified in susequent genome-wide ssocition study. In prticulr, crriers of the minor llele of CDH13 rs11646213 showed decresed risk of hypertension, while crriers of rs3096277 nd rs254340 (locted in intron 11) showed n ssocition with BP. 21 Evidence of the involvement of CDH13 in BP regultion ws demonstrted in nother popultion-sed cohort (Frminghm Hert Study). 21 In hypertension sujects, minor llele crriers of rs12444338 hd lower risk of hypertension, lthough the ssocition ws mrginl fter djusting for confounders. In ddition, the men crotid intim-medi thickness (IMT) ws significntly ssocited with rs12444338 (p=0.02) nd rs1048612 (p=0.02). 22 In this study, genotypes of the SNP rs3865188 were lso ssocited with DBP. Interestingly, we oserved lower insulin levels with incresed diponectin levels in sttin-treted sujects with the T llele, compred to sttin-free groups with the A llele. CDH13 strongly influences circulting diponectin levels nd is ssocited with eneficil metolic profile: the positive reltionship etween diponectin nd insulin sensitivity in Est Asin popultions ppers to e cusl. 23 In the present study, plsm diponectin levels were negtively ssocited with TG nd TG/HDL rtio in ptients with the genotype in the sttinfree group. In the sttin group, however, the effect of the genotype disppered, ecuse diponectin levels ws incresed y sttin dministrtion. We found tht sttin tretment significntly decresed TC, LDLc, TG/HDL rtio, nd HDL3 frction, compred to sttin-free individuls, nd incresed diponectin in persons hving mutnt lleles of the CDH13 gene. In generl, shifting towrd smll-sized HDL ws chnged y serum TG or TC elevtion. Besides, serum TG level is more importnt fctor to chnge the components of smll HDL such s HDL3 or HDL3c, compred to serum TC level. 16 Even though we do not know whether diponectin cuses these lipid normlities nd, thus, whether it is prtly responsile for therogenic risk, high TG/HDL is ssocited with low plsm diponectin concentrtions in nondietic women. 24 In our study, the interction etween CDH13 gene nd sttin my effect HDL supopultions ccording to chnging plsm TG nd TC levels. Mny studies hve reveled tht sttin dministered efore or fter CAD or stroke onset cn decrese mortlity nd enhnce the short-term nd/or long-term outcomes of ischemic stroke. 25,26 However, inflmmtion is not included in the list of clssic risk fctors for therosclerosis, lthough it seems to e n dditionl importnt therpeutic trget for the reduction of crdiovsculr risk. The secretion of iologiclly ctive molecules including MCP-1 hs n ominous impct on therogenesis. Nevertheless, diponectin hs protective effect ginst the development of crdiovsculr disese, nd its levels re reduced in metolic syndrome. 27,28 The use of drugs such s sttin for treting dyslipidemi nd its modifying effects on lipid profiles hs een the focus of recent studies, which suggest tht therpeutic trgeting of inflmmtion my e used s strtegy to decrese crdiovsculr risk. 29 Some reports show tht the use of rosuvsttin, HMG-CoA reductse inhiitor, leds to significnt reduction in crdiovsculr events in individuls without dyslipidemi with elevted high-sensitivity C-rective protein levels, mrker of systemic inflmmtion. 30 In the present study, sttin did not ffect inflmmtory cytokines, such s MCP-1, leptin, visftin, IL-6, nd TNF-α. Menwhile, ptients in the sttin group with the genotype exhiited higher MCP-1 levels thn those in the sttin-free group. Since plsm diponectin level ws incresed y sttin in genotypes, we ssumed tht diponectin nd MCP-1 ws lnced to offset the incresed CVD. The impct of genetic polymorphisms on sttin tretment efficcy hs een nlyzed in severl clinicl trils, nd the vrints in more thn 30 different genes hve een investigted. 15 Even though knowledge otined in this field is quickly growing, results hve not een replicted in lrger ptient groups. 15 Only the APOE4 llele hs een shown to e ssocited with poor response to sttin tretment. Other genes exmined include only APOA5 or cholesterol 7-α hydroxylse. One report detected potentil vrint within the APOE/C1/C3 gene cluster tht could influence sttin tretment efficcy, nd nother pilot study suggested tht the rs4149056 vrint within the SL- CO1B1 gene hd possile sex-dependent effects on sttin tretment efficcy. 31 CDH13 is highly expressed in severl tissues, 1610
Jung Rn Choi, et l. including the hert, ortic wll, neurons of the rin cortex nd spinl cord, nd smll lood vessels, s well s in vriety of cell types, such s vsculr endothelil cells, smooth muscle cells, pericytes, crdiomyocytes, nd cncer cells. 11,32 Reports on cellulr signling hve oserved tht oth LDL nd diponectin re specific lignds for T-cdherin, product of CDH13. The inding of LDL or diponectin to T-cdherin stimultes the nucler fctor-κb signling pthwy, which plys n importnt role in inflmmtion, nd serves s link etween oesity nd vsculr disese. 19,33 In ddition, T-cdherin might modulte vsculr remodeling, neointim formtion, inflmmtion-relted phenomen, nd therosclerosis development y regulting the diponectin levels in the lood nd vrious tissues. 21 In the present study, we showed the contriution of vrints of CDH13 to sttin tretment efficcy. However, the genetic effects of CDH13 rs3865188 nd the complexity of the development of crdiometolic diseses ws not evluted in the present study, ecuse there ws not enough dt to demonstrte n ssocition etween one vrint of CDH13 nd sttin tretment. There re severl limittions of this study. First, we could not find ny reports tht provided n explntion of the ssocition etween the CDH13 polymorphism nd sttin tretment efficcy. There is undnt reserch demonstrting the ssocition of diponectin levels nd the CDH13 gene in severl popultions. However, we oserved tht studies in which genes tht ffect sttin tretment efficcy hve een identified, including the APOE/C1/C3 gene cluster, do not provide sufficient dt to demonstrte the ssocition of CDH13 nd sttin tretment. A similr prolem ws encountered in our study. Second, only one vrint of CDH13 (rs3865188) ws exmined in our study; this is not sufficient to effectively evlute the ssocition etween the CDH13 gene nd diponectin levels. Third, we did not perform ny nlysis to predict the effects of phrmcotherpy, nd the individul efficcy of drug ws determined through genetic nlysis only. In the present study, we found tht the genotypes of CDH13 ply crucil role in regulting diponectin levels with plsm lipid profiles undergoing sttin tretment in Koren CVD nd dyslipidemi ptients. The results of genetic nlyses conducted in the future my help clinicins select the most effective nd sfe tretment for ech individul ptient. Although the economicl nd helth enefits of such pproches re evident, we re still t the erly stges of this reserch nd only eginning to understnd the genetic fctors tht determine sttin tretment efficcy. Nevertheless, the novel vritions within CDH13 my e utilized for etter clinicl mngement of coronry rtery diseses in the future. ACKNOWLEDGEMENTS This study ws funded y the Ministry of Helth nd Welfre (A000385) nd Mid-creer Resercher Progrm through NRF grnt funded y the MEST (2010-0000147), Repulic of Kore. 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