Elsevier

Matrix Biology

Volume 21, Issue 6, October 2002, Pages 487-498
Matrix Biology

MMP13 promoter polymorphism is associated with atherosclerosis in the abdominal aorta of young black males

https://doi.org/10.1016/S0945-053X(02)00053-7Get rights and content

Abstract

Previous studies suggested that remodeling of connective tissue is important in progression of atherosclerosis. We investigated the importance of matrix metalloproteinase 13 (MMP13), in the pathogenesis of atherosclerosis using 995 samples from the Pathobiological Determinants of Atherosclerosis in Youth collection in an association study. We identified two new MMP13 promoter polymorphisms. The genotype for one of the MMP13 polymorphisms was associated with fibrous plaque (P=0.024) in black males. Immunohistochemistry using antibodies for MMP13 showed that MMP13 is expressed in all layers of the aorta. In-vitro transfection experiments with reporter gene constructs and electrophoretic mobility-shift assays showed that the MMP13 polymorphism was a functional variant. MMP13 is therefore, a genetic risk factor for extent of fibrous plaque in the abdominal aorta in young black males. Elucidation of the currently unknown mechanism of the MMP13 polymorphism's action may provide for pharmacological intervention to reduce the severity of atherosclerotic changes in susceptible individuals.

Introduction

Previous studies have shown that matrix metalloproteinases (MMPs) are produced by macrophages in human atherosclerotic lesions and are expressed at increased levels in the lesions (Henney et al., 1991, Galis et al., 1994, Belaaouaj et al., 1995, Brown et al., 1995, Nikkari et al., 1995, Halpert et al., 1996, Li et al., 1996, Sukhova et al., 1999). It is not yet clear if increased or decreased expression of one or more of the MMPs is necessary for the progression from early to late stages of atherosclerosis.

Factors that influence the level of expression of MMPs are likely to be related to the rate of progression in atherosclerosis (Preissner et al., 1999). Some sequence variants in the promoters of MMP genes are capable of influencing the levels of transcription in vitro with a particular variant more active than another variant (Ye et al., 1996, Rutter et al., 1998, Zhang et al., 1999, Henney et al., 2000, Jormsjö et al., 2000). Polymorphisms that alter function are useful for dissecting the role of the respective genes in diseases by genetic association studies (Terwilliger and Göring, 2000). Recently, an association between the severity of coronary atherosclerosis and the MMP9 polymorphism (Zhang et al., 1999), and an association between the luminal diameter of coronary arteries in diabetic patients with atherosclerosis and the MMP12 polymorphism (Jormsjö et al., 2000), were reported. Studies using the MMP3 polymorphism found that the 6A allele was associated with more rapid occlusion of coronary arteries in British (Ye et al., 1995) and Finnish (Humphries et al., 1998) men, and conferred greater risk for clinical events such as restenosis in Dutch males (de Maat et al., 1999). In contrast, the 5A allele conferred greater risk for myocardial infarction (MI) in Japanese males (Terashima et al., 1999).

Atherosclerotic lesions progress through three major stages (Ross, 1993). The earliest recognizable stage is the fatty streak lesion, in which there are characteristically lipid-filled macrophages (foam cells), lipid-containing smooth muscle cells, and extracellular lipid present in the subendothelial space. The fatty streak lesion corresponds to types II and III of the American Heart Association (AHA) microscopic classification of lesions (Stary et al., 1995). The fibrous plaque, in which a fibrous cap comprised of smooth muscle cells and collagen usually covers a central core of lipid, corresponds to type V of the AHA classification. The late stage (type VI) is the complex lesion, in which adherent thrombus or hemorrhage has occurred. Progression through the stages involves deposition and degradation of extracellular matrix (ECM).

We postulated that MMPs are involved in the transition from early to intermediate stages of atherosclerosis since it is accompanied by tissue remodeling. We hypothesized that functional promoter polymorphisms in some MMP genes are correlated with the presence of, or extent of, surface area involved in atherosclerotic lesions at early stages of fibrous plaque formation in the abdominal aorta. Specifically, we identified and tested new polymorphisms in MMP13, a recently identified collagenase.

Section snippets

Identification of novel MMP13 promoter variants

In the MMP13 gene two variants were identified, an 11A/12A insertion/deletion at nt −291 (MMP13v1) and an A to G transition at nt −77 (MMP13v2) (Fig. 1). Both variants had allele frequencies that qualified them as polymorphisms (Terwilliger and Göring, 2000) (Table 1). MMP13v2 occurred in a consensus sequence for the transcription factor PEA3 (A̱GGAAR; underlined nucleotide denotes polymorphic site) (Pendas et al., 1997, Wingender et al., 2001).

Genetic association

We studied a subset of 995 samples from the

Discussion

A number of studies have investigated the role of MMPs in atherosclerosis and MI (Ye et al., 1995, Humphries et al., 1998, de Maat et al., 1999, Terashima et al., 1999) (Zhang et al., 1999, Henney et al., 2000, Jormsjö et al., 2000). The studies have yielded apparently contradictory results in that the less active allele of MMP3 (6A) was associated with more rapid occlusion (Ye et al., 1995, Humphries et al., 1998, de Maat et al., 1999, Henney et al., 2000), but the more active allele (5A) was

Study population

The study was approved by the Wayne State University IRB and the PDAY Steering Committee. The 15 participating PDAY centers collected a total of 2876 autopsy cases with causes of death such as homicides, suicides, and car accidents, to minimize bias (McGill et al., 1995, Strong et al., 1999). Coronary arteries and abdominal aortas were scored for the percent total surface involved in fatty streaks, fibrous plaques, and complicated as well as calcified lesions (Strong et al., 1999). The lesions

Acknowledgements

We thank Dr Arthur W. Zieske for assisting with PDAY informatics. Supported in part by grants from the American Heart Association, Midwest Affiliate (9807761; G.T.), the American Foundation for Aging Research (S. Yoon), the National Center for Human Genome Research (HG01577; J.M.O.) and the National Center for Research Resources (RR03655; J.M.O.).

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