Related Experiment Video
Updated: Apr 23, 2026
HPLC-based Assay to Monitor Extracellular Nucleotide/Nucleoside Metabolism in Human Chronic Lymphocytic Leukemia Cells
Published on: July 20, 2016
Targeting CD13 (aminopeptidase-N) in turn downregulates ADAM17 by internalization in acute myeloid leukaemia cells
Sandrine Bouchet1, Ruoping Tang2, Fanny Fava2
1INSERM U1138, Université Pierre et Marie Curie, Université Paris-Descartes, Centre de Recherche des Cordeliers, Paris, France.
Abstract:
Secreted matrix metalloproteinases (MMP)-2 and MMP-9 and membrane-anchored aminopeptidase-N/CD13 are abnormally expressed in human acute myeloid leukaemia (AML). We previously showed that CD13 ligation by anti-CD13 monoclonal antibodies can induce apoptosis in AML cells. Here, we assessed ADAM17 expression in primary blood blasts CD13+CD33+ from patients with AML. Primary AML cells expressed ADAM17 transcript and its surface expression was higher in subtype M4 (myelomonocytic) and M5 (monocytic) AML specimens than in M0 and M1/M2 (early and granulocytic) specimens. In AML cell lines defining distinct AML subfamilies (HL-60/M2, NB4/M3, THP-1/M5, U937/M5) and primary AML cells cultured ex vivo, anti-CD13 antibodies downregulated surface CD13 and ADAM17 without affecting MMP-2/-9 release. Knockdown of CD13 by siRNA prevented anti-CD13-mediated ADAM17 downregulation, indicating that CD13 is required for ADAM17 downregulation. Soluble ADAM17 was not detected in the medium of anti-CD13 treated cells, suggesting that ADAM17 was not shed. After ligation by anti-CD13, CD13 and ADAM17 were internalized. Subsequently, we found that ADAM17 interacts with CD13. We postulate that the interaction of ADAM17 with CD13 and its downregulation following CD13 engagement has important implications in AML for the known roles of ADAM17 in tumour-associated cell growth, migration and invasion.
Insights
Anti-CD13 antibodies reduce ADAM17 expression in acute myeloid leukaemia (AML) cells by internalizing the protein, suggesting a new therapeutic target. This interaction is crucial for AML cell growth and invasion.
Area of Science:
- Oncology
- Molecular Biology
- Immunology
Background:
- Matrix metalloproteinases (MMP)-2, MMP-9, and aminopeptidase-N/CD13 are abnormally expressed in acute myeloid leukaemia (AML).
- CD13 ligation by anti-CD13 antibodies induces apoptosis in AML cells.
- ADAM17 (a disintegrin and metalloproteinase domain 17) plays roles in tumor growth, migration, and invasion.
Purpose of the Study:
- To assess ADAM17 expression in primary AML cells.
- To investigate the effect of anti-CD13 antibodies on ADAM17 expression and localization.
- To explore the relationship between CD13 and ADAM17 in AML.
Main Methods:
- Analysis of ADAM17 transcript and surface expression in primary AML cells and cell lines.
- Treatment of AML cells with anti-CD13 antibodies and assessment of CD13 and ADAM17 levels.
- CD13 knockdown using siRNA.
- Investigation of ADAM17 shedding and internalization via western blot and immunofluorescence.
- Co-immunoprecipitation to study protein interactions.
Main Results:
- Primary AML cells express ADAM17, with higher surface expression in M4/M5 subtypes.
- Anti-CD13 antibodies downregulate surface CD13 and ADAM17 in AML cell lines and primary cells.
- CD13 knockdown prevents anti-CD13-mediated ADAM17 downregulation.
- ADAM17 is internalized, not shed, after CD13 ligation, and interacts with CD13.
Conclusions:
- CD13 engagement leads to ADAM17 internalization and downregulation, dependent on CD13.
- The interaction between CD13 and ADAM17 has potential implications for controlling AML cell growth, migration, and invasion.
