Colitis-associated cancer is dependent on the interplay between the hemostatic and inflammatory systems and supported by integrin alpha(M)beta(2) engagement of fibrinogen.

A link between colitis and colon cancer is well established, but the mechanisms regulating inflammation in this context are not fully defined. Given substantial evidence that hemostatic system components are powerful modulators of both inflammation and tumor progression, we used gene-targeted mice to directly test the hypothesis that the coagulation factor fibrinogen contributes to colitis-associated colon cancer in mice. This fundamental provisional matrix protein was found to be an important determinant of colon cancer. Fibrinogen deficiency resulted in a dramatic diminution in the number of colonic adenomas formed following azoxymethane/dextran sodium sulfate challenge. More detailed analyses in mice expressing a mutant form of fibrinogen that retains clotting function, but lacks the leukocyte integrin receptor alpha(M)beta(2) binding motif (Fibgamma(390-396A)), revealed that alpha(M)beta(2)-mediated engagement of fibrin(ogen) is mechanistically coupled to local inflammatory processes (e.g., interleukin-6 elaboration) and epithelial alterations that contribute to adenoma formation. Consistent with these findings, the majority of Fibgamma(390-396A) mice developed no discernable adenomas, whereas penetrance was 100% in controls. Furthermore, the adenomas harvested from Fibgamma(390-396A) mice were significantly smaller than those from control mice and less proliferative based on quantitative analyses of mitotic indices, suggesting an additional role for fibrin(ogen) in the growth of established adenomas. These studies show, for the first time, a unique link between fibrin(ogen) and the development of inflammation-driven malignancy. Given the importance of antecedent inflammation in the progression of numerous cancers, these studies suggest that therapies targeting fibrin(ogen)-alpha(M)beta(2) interactions may be useful in preventing and/or treating this important subset of malignancies.

Fibrin(ogen) exacerbates inflammatory joint disease through a mechanism linked to the integrin alphaMbeta2 binding motif.

Fibrin deposition within joints is a prominent feature of arthritis, but the precise contribution of fibrin(ogen) to inflammatory events that cause debilitating joint damage remains unknown. To determine the importance of fibrin(ogen) in arthritis, gene-targeted mice either deficient in fibrinogen (Fib-) or expressing mutant forms of fibrinogen, lacking the leukocyte receptor integrin alphaMbeta2 binding motif (Fibgamma390-396A) or the alphaIIbbeta3 platelet integrin-binding motif (FibgammaDelta5), were challenged with collagen-induced arthritis (CIA). Fib- mice exhibited fewer affected joints and reduced disease severity relative to controls. Similarly, diminished arthritis was observed in Fibgamma390-396A mice, which retain full clotting function. In contrast, arthritis in FibgammaDelta5 mice was indistinguishable from that of controls. Fibrin(ogen) was not essential for leukocyte trafficking to joints, but appeared to be involved in leukocyte activation events. Fib- and Fibgamma390-396A mice with CIA displayed reduced local expression of TNF-alpha, IL-1beta, and IL-6, which suggests that alphaMbeta2-mediated leukocyte engagement of fibrin is mechanistically upstream of the production of proinflammatory mediators. Supporting this hypothesis, arthritic disease driven by exuberant TNF-alpha expression was not impeded by fibrinogen deficiency. Thus, fibrin(ogen) is an important, but context-dependent, determinant of arthritis, and one mechanism linking fibrin(ogen) to joint disease is coupled to alphaMbeta2-mediated inflammatory processes.

Modulation of germ cell apoptosis with a nitric oxide synthase inhibitor in a murine model of congenital cryptorchidism.

PURPOSE: Apoptosis has been implicated in testicular germ cell loss in experimental models of cryptorchidism. Nitric oxide synthase (NOS) has been shown to have a role in apoptosis in many cell types. The Hoxa 11 knockout mouse has congenital bilateral cryptorchidism and is uniformly sterile. We examined the time course of apoptosis in this model and attempted to attenuate this response in vivo by inhibition of NOS. MATERIALS AND METHODS: The offspring of heterozygous Hoxa 11 knockout mice were genotyped by polymerase chain reaction. Homozygous knockout mice treated with the NOS inhibitor Nomega-nitro-L-arginine methyl ester (L-NAME) and untreated controls were sacrificed at weekly intervals at 3 to 13 weeks of age. Spermatogenesis was evaluated with hematoxylin and eosin staining. Germ cell apoptosis was assessed with a TUNEL assay and DNA staining. Co-localization of NOS activity was measured with a polyclonal antibody to endothelial NOS. RESULTS: Impaired spermatogenesis was observed in Hoxa 11 knockout mice. Testis/body weight ratios were decreased in this group at weeks 6 and 7, while body weights were unchanged. Germ cell apoptosis was significantly higher in the knockout group compared to wild-type controls. Co-localization was observed between endothelial NOS activity and apoptotic cells, while mice treated with L-NAME demonstrated improved spermatogenesis and attenuated apoptosis. CONCLUSIONS: Apoptosis and NOS reactivity appeared to co-localize in the seminiferous tubules in the Hoxa 11 knockout mouse model. Treatment with the NOS inhibitor L-NAME attenuated apoptosis and improved spermatogenesis. This finding suggests that early treatment might serve as an adjunct to early surgical intervention to reduce testicular atrophy, although any impact on long-term fertility remains to be determined.