Dual Fluorescence Isogenic Synthetic Lethal Kinase Screen and High-Content Secondary Screening for MUC16/CA125-Selective Agents.

Abstract

Significant strides have been made in the development of precision therapeutics for cancer. Aberrantly expressed glycoproteins represent a potential avenue for therapeutic development. The MUC16/CA125 glycoprotein serves as a biomarker of disease and a driver of malignant transformation in epithelial ovarian cancer. Previously, we demonstrated a proof-of-principle approach to selectively targeting MUC16+ cells. In this report, we performed a synthetic lethal kinase screen using a human kinome RNAi library and identified key pathways preferentially targetable in MUC16+ cells using isogenic dual-fluorescence ovarian cancer cell lines. Using a separate approach, we performed high-content small-molecule screening of six different libraries of 356,982 compounds for MUC16/CA125-selective agents and identified lead candidates that showed preferential cytotoxicity in MUC16+ cells. Compounds with differential activity were selected and tested in various other ovarian cell lines or isogenic pairs to identify lead compounds for structure-activity relationship (SAR) selection. Lead siRNA and small-molecule inhibitor candidates preferentially inhibited invasion of MUC16+ cells in vitro and in vivo, and we show that this is due to decreased activation of MAPK, and non-receptor tyrosine kinases. Taken together, we present a comprehensive screening approach to the development of a novel class of MUC16-selective targeted therapeutics and identify candidates suitable for further clinical development.

• Authors: Thapi D Rao, Mengyao Xu, Stephanie Eng, Guangli Yang, Robin Manson, Nestor Rosales, Raj Kumar, Irva E Veillard, Qin Zhou, Alexia Iasonos, Ouathek Ouerfelli, Hakim Djaballah, David R Spriggs, Oladapo O Yeku
• Journal: Molecular cancer therapeutics (Mol Cancer Ther) Vol. 21 Issue 5 Pg. 775-785 (05 04 2022) ISSN: 1538-8514 [Electronic] United States
• PMID: 35413118 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: ©2022 American Association for Cancer Research.

Chemical References

• CA-125 Antigen
• MUC16 protein, human
• Membrane Proteins

Topics

• CA-125 Antigen (genetics)
• Carcinoma, Ovarian Epithelial (pathology)
• Cell Line, Tumor
• Female
• Fluorescence
• Humans
• Membrane Proteins (metabolism)
• Ovarian Neoplasms (drug therapy, genetics, pathology)