Evaluation of the Anti-Tumor Activity of the Humanized Monoclonal Antibody NEO-201 in Preclinical Models of Ovarian Cancer

Kristen P Zeligs; Maria Pia Morelli; Justin M David; Monica Neuman; Lidia Hernandez; Stephen Hewitt; Michelle Ozaki; Akosua Osei-Tutu; David Anderson; Thorkell Andresson; Sudipto Das; Justin Lack; Abdalla Abdelmaksoud; Massimo Fantini; Philip M Arlen; Kwong Y Tsang; Christina M Annunziata

doi:10.3389/fonc.2020.00805

Evaluation of the Anti-Tumor Activity of the Humanized Monoclonal Antibody NEO-201 in Preclinical Models of Ovarian Cancer

Front Oncol. 2020 Jun 19:10:805. doi: 10.3389/fonc.2020.00805. eCollection 2020.

Authors

Kristen P Zeligs¹, Maria Pia Morelli¹, Justin M David², Monica Neuman¹, Lidia Hernandez¹, Stephen Hewitt¹, Michelle Ozaki¹, Akosua Osei-Tutu¹, David Anderson¹, Thorkell Andresson³, Sudipto Das³, Justin Lack^{4

5}, Abdalla Abdelmaksoud^{5

6}, Massimo Fantini², Philip M Arlen², Kwong Y Tsang², Christina M Annunziata¹

Affiliations

¹ Women's Malignancy Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, United States.
² Precision Biologics, Inc., Bethesda, MD, United States.
³ Protein Characterization Laboratory of the Cancer Research Program (CRTP)/Mass Spectrometry Center, National Institutes of Health, Fredrick, MD, United States.
⁴ NIAID Collaborative Bioinformatics Resource (NCBR), National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States.
⁵ Frederick National Laboratory for Cancer Research, Advanced Biomedical Computational Science, Fredrick, MD, United States.
⁶ Frederick National Laboratory for Cancer Research, CCR Collaborative Bioinformatics Resource, Fredrick, MD, United States.

Abstract

Purpose: Despite high initial response rates with cytoreductive surgery, conventional chemotherapy and the incorporation of biologic agents, ovarian cancer patients often relapse and die from their disease. New approaches are needed to improve patient outcomes. This study was designed to evaluate the antitumor activity of NEO-201 monoclonal antibody (mAb) in preclinical models of ovarian cancer where the NEO-201 target is highly expressed. Experimental Design: Functional analysis of NEO-201 against tumor cell lines was performed by antibody-dependent cellular cytotoxicity (ADCC) assays. Binding of NEO-201 to tumor tissues and cell lines were determined by immunohistochemistry (IHC) and flow cytometry, respectively. Further characterization of the antigen recognized by NEO-201 was performed by mass spectrometry. Ovarian cancer models were used to evaluate the anti-tumor activity of NEO-201 in vivo. NEO-201 at a concentration of 250 g/mouse was injected intraperitoneally (IP) on days 1, 4, and 8. Human PBMCs were injected IP simultaneously as effector cells. Results: Both IHC and flow cytometry revealed that NEO-201 binds prominently to the colon, pancreatic, and mucinous ovarian cancer tissues and cell lines. Immunoprecipitation of the antigen recognized by NEO-201 was performed in human ovarian, colon, and pancreatic cancer cell lines. From these screening, carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5) and CEACAM6 were identified as the most likely targets of NEO-201. Our results confirmed that NEO-201 binds different types of cancers; the binding is highly selective for the tumor cells without cross reactivity with the surrounding healthy tissue. Functional analysis revealed that NEO-201 mediates ADCC killing against human ovarian and colorectal carcinoma cell lines in vitro. In addition, NEO-201 inhibited tumor growth in the presence of activated human PBMCs in orthotopic mouse models of both primary and metastatic ovarian cancer. Importantly, NEO-201 prolonged survival of tumor-bearing mice. Conclusions: These data suggested that NEO-201 has an antitumor activity against tumor cells expressing its antigen. Targeting an antigen expressed in tumors, but not in normal tissues, allows patient selection for optimal treatment. These findings strongly indicate that NEO-201 warrants clinical testing as both a novel therapeutic and diagnostic agent for treatment of ovarian carcinomas. A first in human clinical trial evaluating NEO-201 in adults with chemo-resistant solid tumors is ongoing at the NIH clinical Center.

Keywords: antibody-dependent cellular cytotoxicity; carcinoembryonic antigen-related cell adhesion molecule (CEACAM6); carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5); monoclonal antibody; natural killer cell; tumor-associated antigen.