Development and characterization of carboxy-terminus specific monoclonal antibodies for understanding MUC16 cleavage in human ovarian cancer

Abhijit Aithal, Wade M. Junker, Prakash Kshirsagar, Srustidhar Das, Sukwinder Kaur, Catherine Orzechowski, Shailendra Kumar Gautam, Rahat Jahan, Yuri M. Sheinin, Imayavaramban Lakshmanan, Moorthy Palanimuthu Ponnusamy, Surinder Kumar Batra, Maneesh Jain

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

MUC16 is overexpressed in ovarian cancer and plays important roles in invasion and metastasis. Previously described monoclonal antibodies against cell surface expressed MUC16 recognize the N-terminal tandemly repeated epitopes present in cancer antigen 125 (CA125). MUC16 is cleaved at a specific location, thus, releasing CA125 into the extracellular space. Recent reports have indicated that the retained carboxy-terminal (CT) fragment of MUC16 might play an important role in tumorigenicity in diverse types of cancers. However, limited data is available on the fate and existence of CT fragment on the surface of the cancer cell. Herein, we characterize two monoclonal antibodies (mAbs) showing specificity to the retained juxtamembrane region of MUC16. For the first time, we demonstrate that MUC16 is cleaved in ovarian cancer cells (NIH:OVCAR-3 [OVCAR-3]) and that the cleaved MUC16 subunits remain associated with each other. Immunohistochemical analyses on different grades of ovarian tumor tissues indicated differential reactivity of CA125 and MUC16 CT mAbs. The CA125 (M11) mAb detected 32/40 (80%), while the CT mAb (5E6) detected 33/40 (82.5%) of total ovarian cancer cases. For serous and serous papillary cases, the CA125 (M11) mAb stained 27/31 cases (87%), while CT mAb (5E6) stained 29/31 cases (93.5%). The CT mAb(s) accurately predict expression of MUC16 since their epitopes are not tandemly repeated and their reactivity may not be dependent on O-linked glycosylation. These antibodies can serve as valuable reagents for understanding MUC16 cleavage and may also serve as potential therapeutic agents for treatment of ovarian cancer.

Original languageEnglish (US)
Article numbere0193907
JournalPLoS One
Volume13
Issue number4
DOIs
StatePublished - Apr 1 2018

Fingerprint

ovarian neoplasms
Ovarian Neoplasms
monoclonal antibodies
Monoclonal Antibodies
Antigens
neoplasms
antigens
Neoplasms
Epitopes
epitopes
Cells
Glycosylation
extracellular space
glycosylation
Tumors
metastasis
Antibody Specificity
Tissue
Extracellular Space
Antibodies

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Development and characterization of carboxy-terminus specific monoclonal antibodies for understanding MUC16 cleavage in human ovarian cancer. / Aithal, Abhijit; Junker, Wade M.; Kshirsagar, Prakash; Das, Srustidhar; Kaur, Sukwinder; Orzechowski, Catherine; Gautam, Shailendra Kumar; Jahan, Rahat; Sheinin, Yuri M.; Lakshmanan, Imayavaramban; Palanimuthu Ponnusamy, Moorthy; Batra, Surinder Kumar; Jain, Maneesh.

In: PLoS One, Vol. 13, No. 4, e0193907, 01.04.2018.

Research output: Contribution to journalArticle

Aithal, Abhijit ; Junker, Wade M. ; Kshirsagar, Prakash ; Das, Srustidhar ; Kaur, Sukwinder ; Orzechowski, Catherine ; Gautam, Shailendra Kumar ; Jahan, Rahat ; Sheinin, Yuri M. ; Lakshmanan, Imayavaramban ; Palanimuthu Ponnusamy, Moorthy ; Batra, Surinder Kumar ; Jain, Maneesh. / Development and characterization of carboxy-terminus specific monoclonal antibodies for understanding MUC16 cleavage in human ovarian cancer. In: PLoS One. 2018 ; Vol. 13, No. 4.
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abstract = "MUC16 is overexpressed in ovarian cancer and plays important roles in invasion and metastasis. Previously described monoclonal antibodies against cell surface expressed MUC16 recognize the N-terminal tandemly repeated epitopes present in cancer antigen 125 (CA125). MUC16 is cleaved at a specific location, thus, releasing CA125 into the extracellular space. Recent reports have indicated that the retained carboxy-terminal (CT) fragment of MUC16 might play an important role in tumorigenicity in diverse types of cancers. However, limited data is available on the fate and existence of CT fragment on the surface of the cancer cell. Herein, we characterize two monoclonal antibodies (mAbs) showing specificity to the retained juxtamembrane region of MUC16. For the first time, we demonstrate that MUC16 is cleaved in ovarian cancer cells (NIH:OVCAR-3 [OVCAR-3]) and that the cleaved MUC16 subunits remain associated with each other. Immunohistochemical analyses on different grades of ovarian tumor tissues indicated differential reactivity of CA125 and MUC16 CT mAbs. The CA125 (M11) mAb detected 32/40 (80{\%}), while the CT mAb (5E6) detected 33/40 (82.5{\%}) of total ovarian cancer cases. For serous and serous papillary cases, the CA125 (M11) mAb stained 27/31 cases (87{\%}), while CT mAb (5E6) stained 29/31 cases (93.5{\%}). The CT mAb(s) accurately predict expression of MUC16 since their epitopes are not tandemly repeated and their reactivity may not be dependent on O-linked glycosylation. These antibodies can serve as valuable reagents for understanding MUC16 cleavage and may also serve as potential therapeutic agents for treatment of ovarian cancer.",
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AU - Junker, Wade M.

AU - Kshirsagar, Prakash

AU - Das, Srustidhar

AU - Kaur, Sukwinder

AU - Orzechowski, Catherine

AU - Gautam, Shailendra Kumar

AU - Jahan, Rahat

AU - Sheinin, Yuri M.

AU - Lakshmanan, Imayavaramban

AU - Palanimuthu Ponnusamy, Moorthy

AU - Batra, Surinder Kumar

AU - Jain, Maneesh

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N2 - MUC16 is overexpressed in ovarian cancer and plays important roles in invasion and metastasis. Previously described monoclonal antibodies against cell surface expressed MUC16 recognize the N-terminal tandemly repeated epitopes present in cancer antigen 125 (CA125). MUC16 is cleaved at a specific location, thus, releasing CA125 into the extracellular space. Recent reports have indicated that the retained carboxy-terminal (CT) fragment of MUC16 might play an important role in tumorigenicity in diverse types of cancers. However, limited data is available on the fate and existence of CT fragment on the surface of the cancer cell. Herein, we characterize two monoclonal antibodies (mAbs) showing specificity to the retained juxtamembrane region of MUC16. For the first time, we demonstrate that MUC16 is cleaved in ovarian cancer cells (NIH:OVCAR-3 [OVCAR-3]) and that the cleaved MUC16 subunits remain associated with each other. Immunohistochemical analyses on different grades of ovarian tumor tissues indicated differential reactivity of CA125 and MUC16 CT mAbs. The CA125 (M11) mAb detected 32/40 (80%), while the CT mAb (5E6) detected 33/40 (82.5%) of total ovarian cancer cases. For serous and serous papillary cases, the CA125 (M11) mAb stained 27/31 cases (87%), while CT mAb (5E6) stained 29/31 cases (93.5%). The CT mAb(s) accurately predict expression of MUC16 since their epitopes are not tandemly repeated and their reactivity may not be dependent on O-linked glycosylation. These antibodies can serve as valuable reagents for understanding MUC16 cleavage and may also serve as potential therapeutic agents for treatment of ovarian cancer.

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