Overexpression of Tissue Inhibitor of Metalloproteinases-3 in Intestinal and Cutaneous Lesions of Graft-versus-Host Disease

Abstract

Matrix metalloproteinases (MMPs) have been implicated in the pathobiology of various T-cell-mediated inflammatory disorders of the intestine and skin. Their synthetic inhibitor has been shown to prevent lethal acute graft-versus-host disease in animal models. We intended to determine the expression of MMPs 1, 3, 7, 9, 10, 12, and 19 and tissue inhibitors of metalloproteinases (TIMPs) 1 and 3 in intestinal and cutaneous lesions of patients suffering from graft-versus-host disease after bone marrow transplantation. In situ hybridizations for MMPs 1, 3, 7, 10, and 12 as well as TIMPs 1 and 3 were performed using 35 S-labeled cRNA probes on intestinal (n ‫؍‬ 13) and cutaneous specimens (n ‫؍‬ 9) from patients with graft-versus-host disease. Immunohistochemical stainings were carried out to localize MMP-9, MMP-19, TIMP-3, and TGF-␤1 proteins, and TUNEL staining, to detect apoptotic cells. TIMP-3 mRNA and protein were detected in cutaneous lesions in areas with vacuolar degeneration of the basal epidermal layer in all skin samples, and they colocalized with apoptotic keratinocytes and partly with staining for TGF-␤. None of the MMPs examined were overexpressed in skin lesions. Signals for MMP-1 and MMP-3 mRNA was found in 10/13 and 5/13 intestinal biopsies, respectively. In the gut, MMP-19 -positive epithelial cells, particularly in the crypts, were found in 10/13 samples. Expression of MMPs 7, 9, 10, and 12 was absent or very low. TIMPs 1 and 3 were expressed by stromal cells in 12/13 and 10/13 gut samples, respectively. Whereas TIMP-1 was expressed particularly by subepithelial cells where epithelium had shed away, TIMP-3 was detected in deeper areas. We conclude that MMPs are differentially regulated in the skin and gut lesions of graft-versus-host disease. In agreement with previous data on cancer cells, TIMP-3, induced by TGF-␤1, may contribute to the apoptosis of keratinocytes in cutaneous graftversus-host disease lesions, leading to typical histopathological changes. We also conclude that MMPs play a less important role as effector molecules in intestinal graft-versus-host disease than in celiac or inflammatory bowel disease.

Enhanced metastatic dissemination to multiple organs by melanoma and lymphoma cells in timp-3−/− mice

“…Clinical studies report silencing of TIMP-3 in a number of human cancers, and its reduction in stroma surrounding invasive colorectal cancers (Powe et al, 1997). On the other hand, TIMP-3 is highly expressed in invasive esophageal adenocarcinoma and melanoma (Airola et al, 1999;Salmela et al, 2003). Genetic manipulation of TIMP-3 expression significantly affects the primary tumor at the levels of tumor invasion, angiogenesis and growth.…”

Matrix Metalloproteinase-21 Is Expressed Epithelially During Development and in Cancer and Is Up-Regulated by Transforming Growth Factor-β1 in Keratinocytes

“…The Apoptag in situ apoptosis detection kit (Oncor Inc., Gaithersburg, Maryland) was used to detect fragmented DNA of apoptotic cells (Salmela et al, 2003).…”

Reverse zymography using fluorogenic substrates for protease inhibitor detection

“…novel, high-molecular-mass inhibitor PSTI in the mucosa cell surface of intestine has not been reported in the literature, although some investigations on tissue inhibitors of metalloprotease (TIMP 1 and TIMP 3) were found [20,21]. In this paper, we report only the presence of this inhibitor in the small intestine, and do not attempt to deeply focus on its characteristics.…”