In Vivo Imaging of the Tumor-Associated Enzyme NCEH1 with a Covalent PET Probe

Overview

Journal Angew Chem Int Ed Engl

Specialty Chemistry

Date 2020 May 17

PMID 32415874

Citations 7

Authors

Jae Won Chang

Mohammed Bhuiyan

Hsiu-Ming Tsai

Hannah J Zhang

Gang Li

Shaghayegh Fathi

David C McCutcheon

Lara Leoni

Richard Freifelder

Chin-Tu Chen

Raymond E Moellering

Affiliations

Soon will be listed here.

Abstract

Herein, we report the development of an F-labeled, activity-based small-molecule probe targeting the cancer-associated serine hydrolase NCEH1. We undertook a focused medicinal chemistry campaign to simultaneously preserve potent and specific NCEH1 labeling in live cells and animals, while permitting facile F radionuclide incorporation required for PET imaging. The resulting molecule, [ F]JW199, labels active NCEH1 in live cells at nanomolar concentrations and greater than 1000-fold selectivity relative to other serine hydrolases. [ F]JW199 displays rapid, NCEH1-dependent accumulation in mouse tissues. Finally, we demonstrate that [ F]JW199 labels aggressive cancer tumor cells in vivo, which uncovered localized NCEH1 activity at the leading edge of triple-negative breast cancer tumors, suggesting roles for NCEH1 in tumor aggressiveness and metastasis.

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References

Moellering R, Cravatt B . How chemoproteomics can enable drug discovery and development. Chem Biol. 2012; 19(1):11-22. PMC: 3312051. DOI: 10.1016/j.chembiol.2012.01.001. View

Lentz C, Sheldon J, Crawford L, Cooper R, Garland M, Amieva M . Identification of a S. aureus virulence factor by activity-based protein profiling (ABPP). Nat Chem Biol. 2018; 14(6):609-617. PMC: 6202179. DOI: 10.1038/s41589-018-0060-1. View

Nomura D, Dix M, Cravatt B . Activity-based protein profiling for biochemical pathway discovery in cancer. Nat Rev Cancer. 2010; 10(9):630-8. PMC: 3021511. DOI: 10.1038/nrc2901. View

Li G, Montgomery J, Eckert M, Chang J, Tienda S, Lengyel E . An activity-dependent proximity ligation platform for spatially resolved quantification of active enzymes in single cells. Nat Commun. 2017; 8(1):1775. PMC: 5701173. DOI: 10.1038/s41467-017-01854-0. View

Sanman L, Bogyo M . Activity-based profiling of proteases. Annu Rev Biochem. 2014; 83:249-73. DOI: 10.1146/annurev-biochem-060713-035352. View

Betts H, Milicevic Sephton S, Tong C, Awais R, Hill P, Perkins A . Synthesis, in Vitro Evaluation, and Radiolabeling of Fluorinated Puromycin Analogues: Potential Candidates for PET Imaging of Protein Synthesis. J Med Chem. 2016; 59(20):9422-9430. DOI: 10.1021/acs.jmedchem.6b00968. View

Witney T, James M, Shen B, Chang E, Pohling C, Arksey N . PET imaging of tumor glycolysis downstream of hexokinase through noninvasive measurement of pyruvate kinase M2. Sci Transl Med. 2015; 7(310):310ra169. DOI: 10.1126/scitranslmed.aac6117. View

Lanning B, Whitby L, Dix M, Douhan J, Gilbert A, Hett E . A road map to evaluate the proteome-wide selectivity of covalent kinase inhibitors. Nat Chem Biol. 2014; 10(9):760-767. PMC: 4138289. DOI: 10.1038/nchembio.1582. View

Chang J, Moellering R, Cravatt B . An activity-based imaging probe for the integral membrane hydrolase KIAA1363. Angew Chem Int Ed Engl. 2011; 51(4):966-70. PMC: 3425385. DOI: 10.1002/anie.201107236. View

10.

Chang J, Nomura D, Cravatt B . A potent and selective inhibitor of KIAA1363/AADACL1 that impairs prostate cancer pathogenesis. Chem Biol. 2011; 18(4):476-84. PMC: 3119342. DOI: 10.1016/j.chembiol.2011.02.008. View

11.

Wilson A, Garcia A, Parkes J, Houle S, Tong J, Vasdev N . [11C]CURB: Evaluation of a novel radiotracer for imaging fatty acid amide hydrolase by positron emission tomography. Nucl Med Biol. 2011; 38(2):247-53. DOI: 10.1016/j.nucmedbio.2010.08.001. View

12.

Chen Z, Mori W, Fu H, Schafroth M, Hatori A, Shao T . Design, Synthesis, and Evaluation of F-Labeled Monoacylglycerol Lipase Inhibitors as Novel Positron Emission Tomography Probes. J Med Chem. 2019; 62(19):8866-8872. PMC: 7875603. DOI: 10.1021/acs.jmedchem.9b00936. View

13.

Kim W, Le T, Wei L, Poddar S, Bazzy J, Wang X . [18F]CFA as a clinically translatable probe for PET imaging of deoxycytidine kinase activity. Proc Natl Acad Sci U S A. 2016; 113(15):4027-32. PMC: 4839461. DOI: 10.1073/pnas.1524212113. View

14.

Li G, Moellering R . A Concise, Modular Antibody-Oligonucleotide Conjugation Strategy Based on Disuccinimidyl Ester Activation Chemistry. Chembiochem. 2019; 20(12):1599-1605. PMC: 6773266. DOI: 10.1002/cbic.201900027. View

15.

Okazaki H, Igarashi M, Nishi M, Sekiya M, Tajima M, Takase S . Identification of neutral cholesterol ester hydrolase, a key enzyme removing cholesterol from macrophages. J Biol Chem. 2008; 283(48):33357-64. PMC: 2662263. DOI: 10.1074/jbc.M802686200. View

16.

Marcus C, Mena E, Subramaniam R . Brain PET in the diagnosis of Alzheimer's disease. Clin Nucl Med. 2014; 39(10):e413-22. PMC: 4332800. DOI: 10.1097/RLU.0000000000000547. View

17.

Muir R, Zhao N, Wei J, Wang Y, Moroz A, Huang Y . Measuring Dynamic Changes in the Labile Iron Pool in Vivo with a Reactivity-Based Probe for Positron Emission Tomography. ACS Cent Sci. 2019; 5(4):727-736. PMC: 6487455. DOI: 10.1021/acscentsci.9b00240. View

18.

Li G, Eckert M, Chang J, Montgomery J, Chryplewicz A, Lengyel E . Ultrasensitive, multiplexed chemoproteomic profiling with soluble activity-dependent proximity ligation. Proc Natl Acad Sci U S A. 2019; 116(43):21493-21500. PMC: 6815137. DOI: 10.1073/pnas.1912934116. View

19.

Rashidian M, Keliher E, Bilate A, Duarte J, Wojtkiewicz G, Jacobsen J . Noninvasive imaging of immune responses. Proc Natl Acad Sci U S A. 2015; 112(19):6146-51. PMC: 4434737. DOI: 10.1073/pnas.1502609112. View

20.

Jessani N, Liu Y, Humphrey M, Cravatt B . Enzyme activity profiles of the secreted and membrane proteome that depict cancer cell invasiveness. Proc Natl Acad Sci U S A. 2002; 99(16):10335-40. PMC: 124915. DOI: 10.1073/pnas.162187599. View