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Visualizing mesothelin and mucin interactions: Insights into cancer dynamics

The co-expression of mesothelin and mucin is strongly correlated with a poor prognosis in cancers where these specific proteins are highly expressed. Navinci’s in situ proximity ligation assay reveals this interaction, providing early insights for potential control of tumor migration and invasiveness.

Visualizing mesothelin and mucin interactions: Insights into cancer dynamics

The co-expression of mesothelin and mucin is strongly correlated with a poor prognosis in cancers where these specific proteins are highly expressed. Navinci’s in situ proximity ligation assay reveals this interaction, providing early insights for potential control of tumor migration and invasiveness.

Mesothelin and Mucin interaction in ovary fluorescence readout

Mesothelin and mucin interaction in tissues from malignant ovary showing clear cell carcinoma, detection using NaveniFlex™ MR Atto647N

Mesothelin and mucin – two important proteins in tumorigenesis

Mesothelin is a glycosylphosphatidylinositol (GPI)-anchored cell surface glycoprotein overexpressed in many cancers. Mesothelin expression has been identified in many solid tumors, most robustly in mesothelioma, epithelial ovarian cancer, and pancreatic adenocarcinoma, but also in lung and uterine malignancies as well as cholangiocarcinoma (1,2,3,4,5). Mucin 16 (MUC16) is a cell surface glycoprotein that plays a role in promoting cancer cell growth in ovarian cancer. It is a member of the mucin family of glycoproteins. Studies have shown that MUC16 is overexpressed in multiple tumor types, including malignant mesothelioma, ovarian, breast, pancreatic, and colorectal cancer. Protein-protein interactions between MUC16 and its various receptors may influence cell-cell interactions, allowing cancer cells to evade apoptosis (6, 7, 8, 9 ). The first identified binding site of mucin is localized on mesothelin, detected on mesothelial cells.

The overexpression of MUC16 by many tumors of epithelial origin suggests an important role for mucin in tumorigenesis. The N-terminus of the extracellular domain of mesothelin has specific affinity for MUC16 and by which it mediates cancer cell adhesion and possibly plays a role in the implantation and metastatic spread of tumors (6, 7, 10). The high affinity interaction between the two glycoproteins makes these two molecules excellent therapeutic targets in the effort to prevent and control metastasis (6).

Mesothelin and Mucin interaction in ovary fluorescence readout -2

Mesothelin and mucin interaction in tissues from malignant ovary showing clear cell carcinoma, detection using NaveniFlex™ MR Atto647N

Mesothelin and mucin – two important proteins in tumorigenesis

Mesothelin is a glycosylphosphatidylinositol (GPI)-anchored cell surface glycoprotein overexpressed in many cancers. Mesothelin expression has been identified in many solid tumors, most robustly in mesothelioma, epithelial ovarian cancer, and pancreatic adenocarcinoma, but also in lung and uterine malignancies as well as cholangiocarcinoma (1,2,3,4,5). Mucin 16 (MUC16) is a cell surface glycoprotein that plays a role in promoting cancer cell growth in ovarian cancer. It is a member of the mucin family of glycoproteins. Studies have shown that MUC16 is overexpressed in multiple tumor types, including malignant mesothelioma, ovarian, breast, pancreatic, and colorectal cancer. Protein-protein interactions between MUC16 and its various receptors may influence cell-cell interactions, allowing cancer cells to evade apoptosis (6, 7, 8, 9 ). The first identified binding site of mucin is localized on mesothelin, detected on mesothelial cells.

The overexpression of MUC16 by many tumors of epithelial origin suggests an important role for mucin in tumorigenesis. The N-terminus of the extracellular domain of mesothelin has specific affinity for MUC16 and by which it mediates cancer cell adhesion and possibly plays a role in the implantation and metastatic spread of tumors (6, 7, 10). The high affinity interaction between the two glycoproteins makes these two molecules excellent therapeutic targets in the effort to prevent and control metastasis (6).

Mesothelin and Mucin interaction​

Figure 1. Model of Navinci’s in situ proximity ligation assay for mesothelin and mucin interactions. Only if the Navenibodies are in close proximity will they generate a rolling circle amplification reaction, leading to a strong and distinct signal

Mesothelin and Mucin interaction​

Figure 1. Model of Navinci’s in situ proximity ligation assay for mesothelin and mucin interactions. Only if the Navenibodies are in close proximity will they generate a rolling circle amplification reaction, leading to a strong and distinct signal

Naveni® technology: Illuminating mesothelin-mucin dynamics

Mesothelin and MUC16 are highly expressed and upregulated in several tumor types like epithelial ovarian tumors and pancreatic cancer. Inversely, they have limited expression in normal tissues. We have demonstrated the use of the classic Naveni in situ proximity ligation assay for the visualization of the interaction between mesothelin and MUC16 in FFPE tissues (Figure 1). Given that the co-expression of MUC16 and mesothelin is strongly correlated with a poor prognosis and unfavorable patient outcome (11, 12), this assay is beneficial for visualizing the interaction at an early stage, which may be instrumental in preventing and controlling the migration and invasiveness of tumor cells.​

Naveni® technology: Illuminating mesothelin-mucin dynamics

Mesothelin and MUC16 are highly expressed and upregulated in several tumor types like epithelial ovarian tumors and pancreatic cancer. Inversely, they have limited expression in normal tissues. We have demonstrated the use of the classic Naveni in situ proximity ligation assay for the visualization of the interaction between mesothelin and MUC16 in FFPE tissues (Figure 1). Given that the co-expression of MUC16 and mesothelin is strongly correlated with a poor prognosis and unfavorable patient outcome (11, 12), this assay is beneficial for visualizing the interaction at an early stage, which may be instrumental in preventing and controlling the migration and invasiveness of tumor cells.​

Application example – mesothelin and mucin interactions in ovary brightfield readout

Application example – mesothelin and mucin interactions in ovary brightfield readout

Mesothelin and Mucin interaction clear cell carcinoma ovary A1

Mesothelin and mucin interaction in tissue cores from malignant ovary showing clear cell carcinoma, detection using NaveniBright AP

Mesothelin and Mucin interaction clear cell carcinoma ovary A2

Mesothelin and mucin interaction in tissue cores from malignant ovary showing clear cell carcinoma, detection using NaveniBright AP

Mesothelin and mucin interaction in tissue cores from malignant ovary showing clear cell carcinoma, detection using NaveniBright AP

Application example – mesothelin and mucin interaction in ovary fluorescence readout

Application example – mesothelin and mucin interaction in ovary fluorescence readout

Mesothelin and Mucin interaction in ovary fluorescence readout

Mesothelin and mucin interaction in tissues from malignant ovary showing clear cell carcinoma, detection using NaveniFlex MR Atto647N

Mesothelin and Mucin interaction in ovary fluorescence readout 2

Mesothelin and mucin interaction in tissues from malignant ovary showing clear cell carcinoma, detection using NaveniFlex MR Atto647N

Mesothelin and mucin interaction in tissues from malignant ovary showing clear cell carcinoma, detection using NaveniFlex MR Atto647N

How to detect mesothelin and mucin interactions

Use our products NaveniBright AP and NaveinFlex Tissue MR Atto647N; these products were used in the above examples. For further information about mesothelin/mucin monoclonal antibodies and protocols, contact us using the form below.

How to detect mesothelin and mucin interactions

Use our products NaveniBright AP and NaveinFlex Tissue MR Atto647N; these products were used in the above examples. For further information about mesothelin/mucin monoclonal antibodies and protocols, contact us using the form below.

NaveniBright AP in Ovary

NaveniBright AP

The assay is designed to be used with a mouse and a rabbit primary pair. Chromogenic readout with AP substrate.

NavenFlex Tissue MR_Atto Colon Beta-Catenin_E-Cadherin interaction

NaveniFlex Tissue

The assay is designed to be used with a mouse and rabbit primary antibody pair. Detection with Atto647N fluorophore is included.

NaveniBright AP in Ovary

NaveniBright AP

The assay is designed to be used with a mouse and a rabbit primary pair. Chromogenic readout with AP substrate.

NavenFlex Tissue MR_Atto Colon Beta-Catenin_E-Cadherin interaction

NaveniFlex Tissue

The assay is designed to be used with a mouse and rabbit primary antibody pair. Detection with Atto647N fluorophore is included.

References

  1. Dainty, L. A., Risinger, J. I., Morrison, C., Chandramouli, G. V., Bidus, M. A., Zahn, C., Rose, G. S., Fowler, J., Berchuck, A., & Maxwell, G. L. (2007). Overexpression of folate binding protein and mesothelin are associated with uterine serous carcinoma. Gynecologic oncology105(3), 563–570. DOI: 10.1016/j.ygyno.2006.10.063
  2. Galloway M.L., et al. Histopathology.2006;48:767–769.​
  3. Ho, M., Bera, T. K., Willingham, M. C., Onda, M., Hassan, R., FitzGerald, D., & Pastan, I. (2007). Mesothelin expression in human lung cancer. Clinical cancer research : an official journal of the American Association for Cancer Research13(5), 1571–1575. DOI: 10.1158/1078-0432.CCR-06-2161
  4. Ordóñez N. G. (2003). Application of mesothelin immunostaining in tumor diagnosis. The American journal of surgical pathology27(11), 1418–1428. DOI: 10.1097/00000478-200311000-00003
  5. Yu, L., Feng, M., Kim, H., Phung, Y., Kleiner, D.E., Gores, G.J., Qian, M., Wang, X.W., Ho, M. (2010). Mesothelin as a Potential Therapeutic Target in Human Cholangiocarcinoma. Journal of Cancer, 1, 141-149. DOI: 10.7150/jca.1.141
  6. Gubbels, J. A., Belisle, J., Onda, M., Rancourt, C., Migneault, M., Ho, M., Bera, T. K., Connor, J., Sathyanarayana, B. K., Lee, B., Pastan, I., & Patankar, M. S. (2006). Mesothelin-MUC16 binding is a high affinity, N-glycan dependent interaction that facilitates peritoneal metastasis of ovarian tumors. Molecular cancer5(1), 50. DOI: 10.1186/1476-4598-5-50
  7. Rump, A., Morikawa, Y., Tanaka, M., Minami, S., Umesaki, N., Takeuchi, M., & Miyajima, A. (2004). Binding of ovarian cancer antigen CA125/MUC16 to mesothelin mediates cell adhesion. The Journal of biological chemistry279(10), 9190–9198. DOI: 10.1074/jbc.M312372200
  8. Streppel, M. M., Vincent, A., Mukherjee, R., Campbell, N. R., Chen, S. H., Konstantopoulos, K., Goggins, M. G., Van Seuningen, I., Maitra, A., & Montgomery, E. A. (2012). Mucin 16 (cancer antigen 125) expression in human tissues and cell lines and correlation with clinical outcome in adenocarcinomas of the pancreas, esophagus, stomach, and colon. Human pathology43(10), 1755–1763. DOI: 10.1016/j.humpath.2012.01.005
  9. Shimizu, A., Hirono, S., Tani, M., Kawai, M., Okada, K., Miyazawa, M., Kitahata, Y., Nakamura, Y., Noda, T., Yokoyama, S., & Yamaue, H. (2012). Coexpression of MUC16 and mesothelin is related to the invasion process in pancreatic ductal adenocarcinoma. Cancer science103(4), 739–746. DOI: 10.1111/j.1349-7006.2012.02214.x
  10. Kaneko, O., Gong, L., Zhang, J., Hansen, J. K., Hassan, R., Lee, B., & Ho, M. (2009). A binding domain on mesothelin for CA125/MUC16. The Journal of biological chemistry284(6), 3739–3749. DOI: 10.1074/jbc.M806776200
  11. Belisle, J. A., Horibata, S., Jennifer, G. A., Petrie, S., Kapur, A., André, S., Gabius, H. J., Rancourt, C., Connor, J., Paulson, J. C., & Patankar, M. S. (2010). Identification of Siglec-9 as the receptor for MUC16 on human NK cells, B cells, and monocytes. Molecular cancer9, 118. DOI: 10.1186/1476-4598-9-118