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Updated: Aug 8, 2026
Analysis of Lymph Node Volume by Ultra-High-Frequency Ultrasound Imaging in the Braf/Pten Genetically Engineered Mouse Model of Melanoma
Published on: September 8, 2021
Enhanced metastatic dissemination to multiple organs by melanoma and lymphoma cells in timp-3-/- mice
W Cruz-Munoz1, O H Sanchez, M Di Grappa
1Ontario Cancer Research, University of Toronto, Toronto, Ontario, Canada.
Abstract:
Identifying versatile inhibitors of metastasis that operate in multiple sites against distinct cancer cell types is important for designing novel therapeutics for metastasis. We show that multiple tissues of timp-3-/- mice are more susceptible to metastatic colonization. Overall, a 5-14-fold increase in liver and kidney colonization occurred by EL-4 lymphoma cells, and a twofold increase upon targeting B16F10 melanoma cells to the bone or lung of timp-3-/- mice. There was a general lack of macrophage or neutrophil localization to metastases in the liver, kidney and lung, and of osteoclasts to bone in both genotypes. Analysis of lung showed that proliferation or angiogenesis were unaltered within the metastatic colonies. Lung-trap assays revealed that initial tumor cell trapping was similar in the lung vasculature of timp-3-/- and wild-type mice. However, more tumor cells were found in timp-3-/- lungs at 48 and 96 h after tumor cell injection indicating more efficient extravasation and initial proliferation. Activation of pro-MMP-2 was greater in timp-3-/- lungs at these time points. These data demonstrate TIMP-3 functions to inhibit metastatic dissemination of diverse cancer cells to multiple organs. TIMP-3 regulates MMP-2 activation to limit tumor cell extravasation and subsequent colonization of the lung, without augmenting inflammatory cell response.
Insights
Tissue inhibitor of metalloproteinase-3 (TIMP-3) deficiency increases susceptibility to cancer metastasis in multiple organs. TIMP-3 limits tumor cell extravasation and colonization by regulating matrix metalloproteinase-2 (MMP-2) activation.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Metastasis is a complex process involving tumor cell dissemination, survival, and colonization in distant organs.
- Identifying molecular mechanisms that inhibit metastasis is crucial for developing effective cancer therapies.
- Tissue inhibitor of metalloproteinase-3 (TIMP-3) is a key regulator of extracellular matrix remodeling and has been implicated in cancer progression.
Purpose of the Study:
- To investigate the role of TIMP-3 in inhibiting cancer metastasis to multiple organs.
- To determine how TIMP-3 deficiency affects tumor cell colonization and the tumor microenvironment.
- To elucidate the molecular mechanisms by which TIMP-3 regulates metastasis, focusing on matrix metalloproteinase (MMP) activation.
Main Methods:
- Utilized TIMP-3 knockout (TIMP-3-/-) mice and wild-type littermates.
- Administered EL-4 lymphoma and B16F10 melanoma cells intravenously and via tail vein injection.
- Assessed metastatic colonization in liver, kidney, bone, and lung tissues.
- Performed lung-trap assays to evaluate tumor cell trapping and extravasation.
- Analyzed inflammatory cell infiltration (macrophages, neutrophils, osteoclasts) and evaluated matrix metalloproteinase-2 (MMP-2) activation.
Main Results:
- TIMP-3-/- mice exhibited significantly increased metastatic colonization in the liver, kidney, bone, and lung compared to wild-type mice.
- Tumor cell extravasation and initial proliferation were enhanced in the lungs of TIMP-3-/- mice.
- Pro-MMP-2 activation was elevated in the lungs of TIMP-3-/- mice.
- No significant differences were observed in inflammatory cell infiltration or angiogenesis within metastatic colonies between genotypes.
Conclusions:
- TIMP-3 acts as a critical inhibitor of cancer metastasis across diverse cell types and multiple organs.
- TIMP-3's inhibitory function is mediated through the regulation of MMP-2 activation, thereby limiting tumor cell extravasation and subsequent colonization.
- TIMP-3's role in metastasis inhibition is independent of augmenting inflammatory cell responses.
