Trop-2 cleavage by ADAM10 is an activator switch for cancer growth and metastasis
Marco Trerotola1, Emanuela Guerra1, Zeeshan Ali2
1Laboratory of Cancer Pathology, Center for Advanced Studies and Technology (CAST), University 'G. D'Annunzio', Chieti, Italy; Department of Medical, Oral and Biotechnological Sciences, University 'G. d'Annunzio', Chieti, Italy.
Abstract:
Trop-2 is a transmembrane signal transducer that can induce cancer growth. Using antibody targeting and N-terminal Edman degradation, we show here that Trop-2 undergoes cleavage in the first thyroglobulin domain loop of its extracellular region, between residues R87 and T88. Molecular modeling indicated that this cleavage induces a profound rearrangement of the Trop-2 structure, which suggested a deep impact on its biological function. No Trop-2 cleavage was detected in normal human tissues, whereas most tumors showed Trop-2 cleavage, including skin, ovary, colon, and breast cancers. Coimmunoprecipitation and mass spectrometry analysis revealed that ADAM10 physically interacts with Trop-2. Immunofluorescence/confocal time-lapse microscopy revealed that the two molecules broadly colocalize at the cell membrane. We show that ADAM10 inhibitors, siRNAs and shRNAs abolish the processing of Trop-2, which indicates that ADAM10 is an effector protease. Proteolysis of Trop-2 at R87-T88 triggered cancer cell growth both in vitro and in vivo. A corresponding role was shown for metastatic spreading of colon cancer, as the R87A-T88A Trop-2 mutant abolished xenotransplant metastatic dissemination. Activatory proteolysis of Trop-2 was recapitulated in primary human breast cancers. Together with the prognostic impact of Trop-2 and ADAM10 on cancers of the skin, ovary, colon, lung, and pancreas, these data indicate a driving role of this activatory cleavage of Trop-2 on malignant progression of tumors.
Insights
Trop-2 cleavage between R87-T88, mediated by ADAM10, drives cancer growth and metastasis. This specific proteolysis is absent in normal tissues but prevalent in tumors, highlighting its role in malignant progression.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Trop-2 is a transmembrane protein implicated in cancer progression.
- The precise mechanisms regulating Trop-2's function in tumors remain incompletely understood.
Purpose of the Study:
- To investigate the proteolytic processing of Trop-2.
- To identify the protease responsible for Trop-2 cleavage.
- To elucidate the functional consequences of Trop-2 cleavage in cancer.
Main Methods:
- Antibody targeting and N-terminal Edman degradation to identify cleavage sites.
- Molecular modeling to assess structural changes.
- Coimmunoprecipitation and mass spectrometry to identify interacting proteins.
- In vitro and in vivo assays to evaluate functional impact.
- Analysis of primary human tumors.
Main Results:
- Trop-2 undergoes cleavage between R87 and T88 in its extracellular domain, a process observed in various human tumors but not normal tissues.
- ADAM10 was identified as the effector protease responsible for Trop-2 cleavage, interacting with Trop-2 at the cell membrane.
- Proteolysis of Trop-2 at R87-T88 significantly promoted cancer cell growth in vitro and in vivo, and was crucial for colon cancer metastasis.
- Mutational analysis of the cleavage site abolished metastatic dissemination.
- Activatory proteolysis of Trop-2 was confirmed in primary human breast cancers.
Conclusions:
- ADAM10-mediated cleavage of Trop-2 is a critical event in malignant tumor progression.
- This specific proteolysis drives cancer cell proliferation and metastasis.
- Trop-2 and ADAM10 exhibit prognostic significance across multiple cancer types, suggesting their potential as therapeutic targets.
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