Monomerization of Homodimeric Trefoil Factor 3 (TFF3) by an Aminonitrile Compound Inhibits TFF3-Dependent Cancer Cell Survival

Vijay Pandey; Xi Zhang; Han-Ming Poh; Baocheng Wang; Dukanya Dukanya; Lan Ma; Zhinan Yin; Andreas Bender; Ganga Periyasamy; Tao Zhu; Kanchugarakoppal S Rangappa; Basappa Basappa; Peter E Lobie

doi:10.1021/acsptsci.2c00044

Monomerization of Homodimeric Trefoil Factor 3 (TFF3) by an Aminonitrile Compound Inhibits TFF3-Dependent Cancer Cell Survival

ACS Pharmacol Transl Sci. 2022 Aug 17;5(9):761-773. doi: 10.1021/acsptsci.2c00044. eCollection 2022 Sep 9.

Authors

Vijay Pandey¹, Xi Zhang², Han-Ming Poh³, Baocheng Wang¹, Dukanya Dukanya⁴, Lan Ma^{1

2}, Zhinan Yin^{5

6}, Andreas Bender⁷, Ganga Periyasamy⁸, Tao Zhu^{9

10}, Kanchugarakoppal S Rangappa⁴, Basappa Basappa⁴, Peter E Lobie^{1

2

3}

Affiliations

¹ Tsinghua Berkeley Shenzhen Institute and Institute of Biopharmaceutical and Health Engineering, Tsinghua Shenzhen International Graduate School, Shenzhen 518055, PR China.
² Shenzhen Bay Laboratory, Shenzhen 518055, PR China.
³ Cancer Science Institute of Singapore and Department of Pharmacology, National University of Singapore, Singapore 117599.
⁴ Laboratory of Chemical Biology, Department of Studies in Organic Chemistry, University of Mysore, Manasagangotri, Mysore 570006 Karnataka, India.
⁵ Biomedical Translational Research Institute, Jinan University, 601 Huangpu Avenue West, Guangzhou 510632, PR China.
⁶ Zhuhai Institute of Translational Medicine Zhuhai People's Hospital Affiliated with Jinan University, Jinan University, Zhuhai, Guangdong 519000, PR China.
⁷ Centre for Molecular Informatics, Department of Chemistry, University of Cambridge, Lensfield Road, CB2 1EW Cambridge, United Kingdom.
⁸ DOS in Chemistry, Bangalore University, JB Campus, Bangalore 560001, India.
⁹ Department of Oncology of the First Affiliated Hospital, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui 230027, China.
¹⁰ Hefei National Laboratory for Physical Sciences, the CAS Key Laboratory of Innate Immunity and Chronic Disease, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui 230027, China.

Abstract

Trefoil factor 3 (TFF3) is a secreted protein with an established oncogenic function and a highly significant association with clinical progression of various human malignancies. Herein, a novel small molecule that specifically targets TFF3 homodimeric functions was identified. Utilizing the concept of reversible covalent interaction, 2-amino-4-(4-(6-fluoro-5-methylpyridin-3-yl)phenyl)-5-oxo-4H,5H-pyrano[3,2-c]chromene-3-carbonitrile (AMPC) was identified as a molecule that interacted with TFF3. AMPC monomerized the cellular and secreted TFF3 homodimer at the cysteine (Cys)57-Cys57 residue with subsequent more rapid degradation of the generated TFF3 monomers. Hence, AMPC treatment also resulted in cellular depletion of TFF3 with consequent decreased cell viability in various human carcinoma-derived TFF3 expressing cell lines, including estrogen receptor positive (ER+) mammary carcinoma (MC). AMPC treatment of TFF3 expressing ER+ MC cells significantly suppressed total cell number in a dose-dependent manner. Consistently, exposure of TFF3 expressing ER+ MC cells to AMPC decreased soft agar colony formation, foci formation, and growth in suspension culture and inhibited growth of preformed colonies in 3D Matrigel. AMPC increased apoptosis in TFF3 expressing ER+ MC cells associated with decreased activity of EGFR, p38, STAT3, AKT, and ERK, decreased protein levels of CCND1, CCNE1, BCL2, and BCL-XL, and increased protein levels of TP53, CDKN1A, CASP7, and CASP9. siRNA-mediated depletion of TFF3 expression in ER+ MC cells efficiently abrogated AMPC-stimulated loss of cell viability and CASPASE 3/7 activities. Furthermore, in mice bearing ER+ MC cell-generated xenografts, AMPC treatment significantly impeded xenograft growth. Hence, AMPC exemplifies a novel mechanism by which small molecule drugs may inhibit a dimeric oncogenic protein and provides a strategy to impede TFF3-dependent cancer progression.