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Macrophage-specific overexpression of human matrix metalloproteinase-12 in transgenic rabbits
Jianglin Fan1, Xiaofei Wang, Lihua Wu
1Laboratory of Cardiovascular Disease, Department of Pathology, Institute of Basic Medical Sciences, University of Tsukuba, Tsukuba 305-8575, Japan. j-lfan@md.tsukuba.ac.jp
Abstract:
Increased matrix metalloproteinase-12 (MMP-12) has been implicated in atherosclerosis and many other inflammatory processes. To define MMP-12 functions in vivo, we generated transgenic rabbits that expressed human (h) MMP-12 gene under the control of a macrophage-specific promoter, the human scavenger receptor promoter. Two transgenic founder rabbits were found to have hMMP-12 transgene integration by Southern blot analysis. hMMP-12 mRNA was expressed in peritoneal and alveolar macrophages, and in tissues enriched in macrophages in transgenic rabbits. High levels of hMMP-12 protein were detected in the conditioned media of cultured peritoneal and alveolar macrophages from transgenic rabbits. Zymography showed that hMMP-12 secreted from macrophages possessed enzymatic activity toward beta-casein. To evaluate the expression of hMMP-12 in inflammatory sites, we used carrageenan-induced granulomas as an in vivo model for tissue macrophages and foam cells. Granuloma size in transgenic rabbits was significantly increased compared to that in control rabbits, and histological examination revealed that granulomas of transgenic rabbits were enriched in macrophages associated with increased hMMP-12 expression. We believe that this transgenic rabbit model with increased expression of hMMP-12 may become a useful model for further mechanistic studies of MMP-12 in inflammatory diseases and cancer invasion; it is also an ideal model for testing the in vivo action of MMP-12 inhibitors.
Insights
Transgenic rabbits with increased matrix metalloproteinase-12 (MMP-12) expression showed larger granulomas, indicating MMP-12 promotes inflammation. This model aids in studying inflammatory diseases and testing MMP-12 inhibitors.
Area of Science:
- Biochemistry
- Immunology
- Genetics
Background:
- Matrix metalloproteinase-12 (MMP-12) is linked to atherosclerosis and inflammatory conditions.
- Understanding MMP-12's in vivo function is crucial for developing targeted therapies.
Purpose of the Study:
- To create and characterize a transgenic rabbit model with enhanced macrophage-specific expression of human MMP-12 (hMMP-12).
- To investigate the role of elevated MMP-12 in inflammatory processes using an in vivo model.
Main Methods:
- Generated transgenic rabbits expressing hMMP-12 under a macrophage-specific promoter (human scavenger receptor promoter).
- Confirmed transgene integration via Southern blot and assessed hMMP-12 mRNA and protein expression in macrophages and tissues.
- Utilized carrageenan-induced granulomas as an in vivo model to evaluate MMP-12's effect on inflammation.
Main Results:
- Transgenic rabbits exhibited hMMP-12 expression in macrophages and macrophage-rich tissues.
- Secreted hMMP-12 from macrophages demonstrated enzymatic activity.
- Carrageenan-induced granulomas were significantly larger in transgenic rabbits, with increased macrophage infiltration and hMMP-12 expression.
Conclusions:
- The developed transgenic rabbit model effectively demonstrates increased MMP-12 expression in inflammatory sites.
- Elevated MMP-12 activity contributes to the exacerbation of inflammatory responses, as evidenced by increased granuloma size.
- This model is valuable for mechanistic studies of MMP-12 in inflammatory diseases, cancer invasion, and for evaluating MMP-12 inhibitors in vivo.
