Ischemia-induced cleavage of cadherins in NRK cells requires MT1-MMP (MMP-14)

ArticleinAmerican journal of physiology. Renal physiology 290(1):F43-51 · February 2006with8 Reads
DOI: 10.1152/ajprenal.00179.2005 · Source: PubMed
Abstract
Ischemia is a leading cause of acute renal failure (ARF), a disease associated with high morbidity and mortality. Disruption of intercellular adhesion in the proximal tubules is linked to ARF, although the molecular mechanism(s) remains unclear. Our previous studies showed that ischemia is associated with cadherin cleavage and loss in NRK cells, putatively due to a matrix metalloproteinase (MMP) (7). In the current studies, a MMP required for E-cadherin cleavage and N-cadherin loss was identified. Chemical inhibitors against a number of soluble MMPs (1, 2, 3, 8, 9) failed to completely attenuate ischemia-induced cadherin loss. Under ischemic conditions, there was an increase in active membrane-type (MT)1-MMP but a decrease in MMP-2 protein expression. Plating cells on fibronectin protected against ischemia-induced loss of cadherins and, interestingly, no increase in active MT1-MMP levels was seen in ischemic cells on fibronectin-coated dishes. In addition, L cells stably expressing E- (LE) or N-cadherin (LN), but lacking MT1-MMP expression, were resistant to ischemia-induced cadherin loss. The role of MT1-MMP in ischemia-induced cadherin loss was confirmed by either blocking MT1-MMP activity with a neutralizing antibody or expression with shRNA constructs which protected full-length E- and N-cadherin during ischemia. Using shRNA constructs to suppress MT1-MMP expression, ischemia-induced disruption of cadherin function was ablated, and cell-cell contacts were preserved. These results demonstrate that ischemia induces increased expression of active MT1-MMP and subsequent disruption of cadherin/catenin complexes, implying that MT1-MMP plays a role in ischemia-induced ARF.
    • "Western assays revealed that E-cadherin is produced in JEG-3 epithelial cells and NRK-49F fibroblasts, although in the latter case it appeared mostly as processed forms (Fig. S1, supplemental material). Processing of E-cadherin by metallo-proteinases (MMPs) has been reported in pathophysiological processes (Covington et al., 2006). Of interest, NRK-49F fibroblasts are also known to produce MMPs in culture (Rankin et al., 2013 ). "
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    • "In the frog gastrula, increased cell–matrix interactions are necessary to suppress inappropriate membrane protrusive activity (Davidson et al., 2006). It is also important to note that Mmp14 has non-ECM protein cleavage substrates capable of impacting cell migration including cadherins (Covington et al., 2006). Our data indicate that vangl2 mutant embryos have decreased membrane cadherin protein expression and future work will determine whether this is due to increased Mmp14 proteolytic activity. "
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