Metastasis is strongly reduced by the matrix metalloproteinase inhibitor Galardin in the MMTV-PymT transgenic breast cancer model

Abstract

Matrix metalloproteinases (MMP) have several roles that influence cancer progression and dissemination. However, low molecular weight metalloproteinase inhibitors (MPI) have not yet been tested in transgenic/spontaneous metastasis models. We have tested Galardin/GM6001, a potent MPI that reacts with most MMPs, in the MMTVPymT transgenic breast cancer model. We followed a cohort of 81 MMTV-PymT transgenic mice that received Galardin, placebo, or no treatment. Galardin treatment was started at age 6 weeks with 100 mg/kg/d, and all mice were killed at age 13.5 weeks. Galardin treatment significantly reduced primary tumor growth. Final tumor burden in Galardin-treated mice was 1.69 cm 3 compared with 3.29 cm 3 in placebo-treated mice (t test, P = 0.0014). We quantified the total lung metastasis volume in the same cohort of mice. The median metastasis volume was 0.003 mm 3 in Galardin-treated mice compared with 0.56 mm 3 in placebo-treated mice (t test, P < 0.0001). Thus, metastasis burden was reduced more than 100-fold, whereas primary tumor size was reduced only 2-fold. We also found that primary tumors from Galardin-treated mice exhibited a lower histopathologic tumor grade, increased collagen deposition, and increased MMP-2 activity. MMPs are known to have tumor-promoting and tumor-inhibitory effects, and several clinical trials of broad-spectrum MPIs have failed to show promising effects. The very potent antimetastatic effect of Galardin in the MMTV-PymT model does, however, show that it may be possible to find broad-spectrum MPIs with favorable inhibition profiles, or perhaps combinations of monospecific MPIs, for future clinical application. [Mol Cancer Ther 2008;7(9):2758 -67]

Effect of a nutrient mixture on the localization of extracellular matrix proteins in HeLa human cervical cancer xenografts in female nude mice

“…In marked contrast, NM supplementation induced intense internal collagen IV production, forming a dense fibrous network of collagen IV that surrounded live nucleated cells and large amounts of necrotic cellular debris. The fibrotic response observed with NM supplementation is similar to the collagen deposition reported by Almholt et al when investigating an experimental pan-MMP inhibiting drug, which was shown to decrease metastatic burden 100-fold (21). Furthermore, tumors from the control group mice exhibited little to no collagen I expression either internally or in the fibrous capsule.…”

Migration of breast cancer cells into reconstituted type I collagen gels assessed via a combination of frozen sectioning and azan staining

“…Additionally, this approach is suited to observing the promotion of cell migration by chemoattractants ( Figure 2) and to assessing the inhibition of cell migration by pharmaceuticals such as galardin. Galardin is a known as MMP inhibitor (11)(12)(13) and is reported to prevent invasion and metastasis (14,15) (Figure 3).…”

“…Recently, various quantitative methods of assessing cell migration and invasion have been reported (8,(14)(15)(16)(17)(18)(19). Some of these methods facilitate the fl uorescenceassisted quantification or real-time observation of migrating cells.…”

The Treatment of Breast Cancer Tumor Growth and Metastasis With an Anti-MMP9 DNAzyme

“…In addition, the lung tumors developed in MMTV-PyMT MMP-9 wild type mice are larger in size and are more highly vascular compared to those tumors that developed in MMTV-PyMT MMP-9 null mice. Thus, the data presented here is consistent with the above studies and demonstrate that down regulation of MMP-9 by AM9D also affects tumor growth via inhibiting angiogenesis and inducing apoptosis ( Our results are consistent with those of Almholt, et al [67] which showed that the broad-spectrum MMP inhibitor Galardin/GM6001, significantly reduced primary mammary tumor growth by 50% and reduced lung metastasis by more than 100-fold in the MMTV-PyMT model. However, contrary to broad-spectrum MMP inhibitors, including GM6001, AM9D treatment specifically down regulates MMP-9 without affecting the expression of other members of the MMP family ( Figure 1-3b and 1-3c).…”

“…More than 90% of mice develop pulmonary metastasis by 100 days [62]. More importantly, the mammary adenocarcinomas exhibit changes in biomarkers and expression patterns of various MMPs similar to those observed in human breast cancer patients [59,[63][64][65][66][67]. Therefore, this model was chosen to ascertain the role of AM9D as a pharmacologic inhibitor of MMP-9.…”