An activatable NIR fluorescent rosol for selectively imaging nitroreductase activity
Abstract
Hypoxia (i.e., pO2 ≤ ~1.5 %) is an important characteristic of tumor microenvironments that directly correlates with resistance against first-line therapies and tumor proliferation/infiltration. The ability to accurately identify hypoxic tumor cells/tissue could afford tailored therapeutic regimens for personalized treatment, the development of more-effective therapies, and discerning the mechanisms underlying disease progression. Fluorogenic constructs currently developed for identifying such tumor cells/tissue operate by targeting the bioreductive activity of primarily the nitroreductase (NTR) class of enzymes, but collectively such constructs unfortunately present photophysical and/or physicochemical shortcomings that could limit their effectiveness upon implementation. To overcome these limitations, we present the rational design, development, and evaluation of the first activatable ultracompact xanthene core-based molecular probe (NO2-Rosol) for selectively imaging NTR activity that affords an “OFF-ON” near-infrared (NIR) fluorescence response (ca. > 700 nm) alongside a remarkable Stokes shift (ca. > 150 nm) via NTR activity-facilitated modulation to its energetics whereby the resultant interplay discontinues an intramolecular d-PET fluorescence-quenching mechanism transpiring between directly-linked electronically-uncoupled π-systems comprising its components. DFT calculations guided selection of a suitable fluorogenic scaffold and nitroaromatic moiety candidate that when adjoined could (i) afford such photophysical response upon bioreduction by upregulated NTR activity in hypoxic tumor cells/tissue and (ii) employmore »
- Authors:
-
- Stanford Univ., CA (United States)
- Publication Date:
- Research Org.:
- Stanford Univ., CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC); National Institutes of Health (NIH)
- OSTI Identifier:
- 1801819
- Alternate Identifier(s):
- OSTI ID: 1582473
- Grant/Contract Number:
- SC0008397; R21 CA205564-01A1; T32 CA118681
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Sensors and Actuators. B, Chemical
- Additional Journal Information:
- Journal Volume: 306; Journal Issue: C; Journal ID: ISSN 0925-4005
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; chemistry; electrochemistry; instruments & instrumentation; hypoxia; fluorescence; imaging; nitroreductase; activatable probe; near-infrared
Citation Formats
Klockow, Jessica L., Hettie, Kenneth S., LaGory, Edward L., Moon, Eui Jung, Giaccia, Amato J., Graves, Edward E., and Chin, Frederick T. An activatable NIR fluorescent rosol for selectively imaging nitroreductase activity. United States: N. p., 2020.
Web. doi:10.1016/j.snb.2019.127446.
Klockow, Jessica L., Hettie, Kenneth S., LaGory, Edward L., Moon, Eui Jung, Giaccia, Amato J., Graves, Edward E., & Chin, Frederick T. An activatable NIR fluorescent rosol for selectively imaging nitroreductase activity. United States. https://doi.org/10.1016/j.snb.2019.127446
Klockow, Jessica L., Hettie, Kenneth S., LaGory, Edward L., Moon, Eui Jung, Giaccia, Amato J., Graves, Edward E., and Chin, Frederick T. Mon .
"An activatable NIR fluorescent rosol for selectively imaging nitroreductase activity". United States. https://doi.org/10.1016/j.snb.2019.127446. https://www.osti.gov/servlets/purl/1801819.
@article{osti_1801819,
title = {An activatable NIR fluorescent rosol for selectively imaging nitroreductase activity},
author = {Klockow, Jessica L. and Hettie, Kenneth S. and LaGory, Edward L. and Moon, Eui Jung and Giaccia, Amato J. and Graves, Edward E. and Chin, Frederick T.},
abstractNote = {Hypoxia (i.e., pO2 ≤ ~1.5 %) is an important characteristic of tumor microenvironments that directly correlates with resistance against first-line therapies and tumor proliferation/infiltration. The ability to accurately identify hypoxic tumor cells/tissue could afford tailored therapeutic regimens for personalized treatment, the development of more-effective therapies, and discerning the mechanisms underlying disease progression. Fluorogenic constructs currently developed for identifying such tumor cells/tissue operate by targeting the bioreductive activity of primarily the nitroreductase (NTR) class of enzymes, but collectively such constructs unfortunately present photophysical and/or physicochemical shortcomings that could limit their effectiveness upon implementation. To overcome these limitations, we present the rational design, development, and evaluation of the first activatable ultracompact xanthene core-based molecular probe (NO2-Rosol) for selectively imaging NTR activity that affords an “OFF-ON” near-infrared (NIR) fluorescence response (ca. > 700 nm) alongside a remarkable Stokes shift (ca. > 150 nm) via NTR activity-facilitated modulation to its energetics whereby the resultant interplay discontinues an intramolecular d-PET fluorescence-quenching mechanism transpiring between directly-linked electronically-uncoupled π-systems comprising its components. DFT calculations guided selection of a suitable fluorogenic scaffold and nitroaromatic moiety candidate that when adjoined could (i) afford such photophysical response upon bioreduction by upregulated NTR activity in hypoxic tumor cells/tissue and (ii) employ a retention mechanism strategy that capitalizes on an inherent physical property of the NIR fluorogenic scaffold for achieving signal amplification. NO2-Rosol demonstrated 705 nm NIR fluorescence emission and a 157 nm Stokes shift, selectivity for NTR over relevant bioanalytes, and a 28- and 12-fold fluorescence enhancement in solution and between cells cultured under different oxic conditions, respectively. Finally, in establishing feasibility for NO2-Rosol to provide favorable contrast levels in in solutioand in vitrostudies, we anticipate NO2-Rosol similarly doing so in future studies towards its translation.},
doi = {10.1016/j.snb.2019.127446},
journal = {Sensors and Actuators. B, Chemical},
number = C,
volume = 306,
place = {United States},
year = {Mon Nov 30 00:00:00 EST 2020},
month = {Mon Nov 30 00:00:00 EST 2020}
}
Web of Science
Works referenced in this record:
Single Amino Acid Switch between a Flavin-Dependent Dehalogenase and Nitroreductase
journal, December 2015
- Mukherjee, Arnab; Rokita, Steven E.
- Journal of the American Chemical Society, Vol. 137, Issue 49
Characterization of chemical, radiochemical and optical properties of a dual-labeled MMP-9 targeting peptide
journal, June 2011
- Azhdarinia, Ali; Wilganowski, Nathaniel; Robinson, Holly
- Bioorganic & Medicinal Chemistry, Vol. 19, Issue 12
Ultrasensitive Near-Infrared Fluorescence-Enhanced Probe for in Vivo Nitroreductase Imaging
journal, May 2015
- Li, Yuhao; Sun, Yun; Li, Jiachang
- Journal of the American Chemical Society, Vol. 137, Issue 19
Resting-State BOLD MRI for Perfusion and Ischemia
journal, January 2017
- Kroll, Hannes; Zaharchuk, Greg; Christen, Thomas
- Topics in Magnetic Resonance Imaging, Vol. 26, Issue 2
Steady-state kinetic mechanism, stereospecificity, substrate and inhibitor specificity of Enterobacter cloacae nitroreductase
journal, September 1998
- Koder, Ronald L.; Miller, Anne-Frances
- Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, Vol. 1387, Issue 1-2
Two-photon fluorescent probe for detection of nitroreductase and hypoxia-specific microenvironment of cancer stem cell
journal, September 2018
- Liu, Yajing; Liu, Wei; Li, Hongjuan
- Analytica Chimica Acta, Vol. 1024
NQO1 overexpression is associated with poor prognosis in squamous cell carcinoma of the uterine cervix
journal, June 2014
- Ma, Yue; Kong, Jienan; Yan, Guanghai
- BMC Cancer, Vol. 14, Issue 1
Clinical implications of high NQO1 expression in breast cancers
journal, January 2014
- Yang, Yang; Zhang, Yan; Wu, Qunying
- Journal of Experimental & Clinical Cancer Research, Vol. 33, Issue 1
Nitroreductase Detection and Hypoxic Tumor Cell Imaging by a Designed Sensitive and Selective Fluorescent Probe, 7-[(5-Nitrofuran-2-yl)methoxy]-3 H -phenoxazin-3-one
journal, March 2013
- Li, Zhao; Li, Xiaohua; Gao, Xinghui
- Analytical Chemistry, Vol. 85, Issue 8
High levels of expression of the NAD(P)H:Quinone oxidoreductase (NQO1) gene in tumor cells compared to normal cells of the same origin
journal, August 1991
- Cresteil, Thierry; Jaiswal, Anil K.
- Biochemical Pharmacology, Vol. 42, Issue 5
The Clinical Importance of Assessing Tumor Hypoxia: Relationship of Tumor Hypoxia to Prognosis and Therapeutic Opportunities
journal, October 2014
- Walsh, Joseph C.; Lebedev, Artem; Aten, Edward
- Antioxidants & Redox Signaling, Vol. 21, Issue 10
Turn-On Near-Infrared Fluorescent Sensor for Selectively Imaging Serotonin
journal, October 2015
- Hettie, Kenneth S.; Glass, Timothy E.
- ACS Chemical Neuroscience, Vol. 7, Issue 1
Selective Catecholamine Recognition with NeuroSensor 521: A Fluorescent Sensor for the Visualization of Norepinephrine in Fixed and Live Cells
journal, March 2013
- Hettie, Kenneth S.; Liu, Xin; Gillis, Kevin D.
- ACS Chemical Neuroscience, Vol. 4, Issue 6
Immunohistochemical Reevaluation of Carbonic Anhydrase IX (CA IX) Expression in Tumors and Normal Tissues
journal, February 2014
- Luong-Player, Adelina; Liu, Haiyan; Wang, Hanlin L.
- American Journal of Clinical Pathology, Vol. 141, Issue 2
Near-Infrared Fluorescent Rosol Dye Tailored toward Lymphatic Mapping Applications
journal, January 2019
- Hettie, Kenneth S.; Klockow, Jessica L.; Glass, Timothy E.
- Analytical Chemistry, Vol. 91, Issue 4
Red-Emitting Fluorescent Probe for Detection of γ-Glutamyltranspeptidase and Its Application of Real-Time Imaging under Oxidative Stress in Cells and in Vivo
journal, May 2018
- Liu, Feiyan; Wang, Zhen; Wang, Wenli
- Analytical Chemistry, Vol. 90, Issue 12
Targeting NAD(P)H:Quinone Oxidoreductase (NQO1) in Pancreatic Cancer
journal, March 2006
- Lewis, Anne M.; Ough, Matthew; Du, Juan
- Molecular Carcinogenesis, Vol. 56, Issue 7
PnrA, a new nitroreductase-family enzyme in the TNT-degrading strain Pseudomonas putida JLR11
journal, August 2005
- Caballero, Antonio; Lazaro, Juan J.; Ramos, Juan L.
- Environmental Microbiology, Vol. 7, Issue 8
Synthesis and characterization of new rhodamine dyes with large Stokes shift
journal, December 2013
- Tian, Zhidan; Tian, Baozhu; Zhang, Jinlong
- Dyes and Pigments, Vol. 99, Issue 3
Molecular Imaging of Hypoxia
journal, June 2008
- Krohn, Kenneth A.; Link, Jeanne M.; Mason, Ralph P.
- Journal of Nuclear Medicine, Vol. 49, Issue Suppl 2
Highly selective and controllable synthesis of arylhydroxylamines by the reduction of nitroarenes with an electron-withdrawing group using a new nitroreductase BaNTR1
journal, January 2014
- Nguyen-Tran, Hieu-Huy; Zheng, Gao-Wei; Qian, Xu-Hong
- Chemical Communications, Vol. 50, Issue 22
Cytochrome P450 CYP3A4/5 Expression as a Biomarker of Outcome in Osteosarcoma
journal, July 2003
- Dhaini, Hassan R.; Thomas, Dafydd G.; Giordano, Thomas J.
- Journal of Clinical Oncology, Vol. 21, Issue 13
Bioreductive fluorescent imaging agents: applications to tumour hypoxia
journal, January 2016
- Elmes, Robert B. P.
- Chemical Communications, Vol. 52, Issue 58
Hypoxia-Sensitive Fluorescent Probes for in Vivo Real-Time Fluorescence Imaging of Acute Ischemia
journal, November 2010
- Kiyose, Kazuki; Hanaoka, Kenjiro; Oushiki, Daihi
- Journal of the American Chemical Society, Vol. 132, Issue 45
NQO1 inhibits proteasome-mediated degradation of HIF-1α
journal, December 2016
- Oh, Eun-Taex; Kim, Jung-whan; Kim, Joon Mee
- Nature Communications, Vol. 7, Issue 1
Design of Protease Activated Optical Contrast Agents That Exploit a Latent Lysosomotropic Effect for Use in Fluorescence-Guided Surgery
journal, June 2015
- Ofori, Leslie O.; Withana, Nimali P.; Prestwood, Tyler R.
- ACS Chemical Biology, Vol. 10, Issue 9
Why is the partial oxygen pressure of human tissues a crucial parameter? Small molecules and hypoxia
journal, May 2011
- Carreau, Aude; Hafny-Rahbi, Bouchra El; Matejuk, Agata
- Journal of Cellular and Molecular Medicine, Vol. 15, Issue 6
Exploiting the Drug-Activating Properties of a Novel Trypanosomal Nitroreductase
journal, March 2010
- Hall, Belinda S.; Wu, Xinghua; Hu, Longqin
- Antimicrobial Agents and Chemotherapy, Vol. 54, Issue 3
Multifunctional Probe Based on Cationic Conjugated Polymers for Nitroreductase-Related Analysis: Sensing, Hypoxia Diagnosis, and Imaging
journal, April 2017
- Zhang, Xiaoqian; Zhao, Qi; Li, Yanru
- Analytical Chemistry, Vol. 89, Issue 10
Development of Unique Xanthene-Cyanine Fused Near-Infrared Fluorescent Fluorophores with Superior Chemical Stability for Biological Fluorescence Imaging
journal, November 2014
- Chen, Hua; Lin, Weiying; Cui, Haijun
- Chemistry - A European Journal, Vol. 21, Issue 2
Hypoxia-related molecules HIF-1α, CA9, and osteopontin: Predictors of survival in patients with high-grade glioma
journal, December 2012
- Erpolat, O. P.; Gocun, P. U.; Akmansu, M.
- Strahlentherapie und Onkologie, Vol. 189, Issue 2
Coumarin-3-Aldehyde as a Scaffold for the Design of Tunable PET-Modulated Fluorescent Sensors for Neurotransmitters
journal, October 2014
- Hettie, Kenneth S.; Glass, Timothy E.
- Chemistry - A European Journal, Vol. 20, Issue 52
Measuring Hypoxia in Solid Tumours&Is There a Gold Standard?
journal, January 2001
- L. Olive, Judit P. Banáth, Christin, Peggy
- Acta Oncologica, Vol. 40, Issue 8
Cytochrome P450 CYP1B1 over-expression in primary and metastatic ovarian cancer
journal, July 2001
- McFadyen, M. C. E.; Cruickshank, M. E.; Miller, I. D.
- British Journal of Cancer, Vol. 85, Issue 2
Expression of NAD(P)H:Quinone Oxidoreductase 1 in HeLa Cells
journal, November 2001
- Bello, Rosario I.; Gómez-Dı́az, Consuelo; Navarro, Francisco
- Journal of Biological Chemistry, Vol. 276, Issue 48
Efficient Two-Photon Fluorescent Probe for Nitroreductase Detection and Hypoxia Imaging in Tumor Cells and Tissues
journal, November 2015
- Zhang, Jing; Liu, Hong-Wen; Hu, Xiao-Xiao
- Analytical Chemistry, Vol. 87, Issue 23
Carbonic anhydrase IX is a clinically significant tissue and serum biomarker associated with renal cell carcinoma
journal, October 2012
- Takacova, Martina; Bartosova, Maria; Skvarkova, Lucia
- Oncology Letters, Vol. 5, Issue 1
Immunodetection of NAD(P)H:quinone oxidoreductase 1 (NQO1) in human tissues11This work is dedicated to the memory of Professor Lars Ernster, who provided us with enthusiastic support, scientific insight, and constant encouragement in our many interactions.
journal, August 2000
- Siegel, David; Ross, David
- Free Radical Biology and Medicine, Vol. 29, Issue 3-4
A Molecular Imaging Primer: Modalities, Imaging Agents, and Applications
journal, April 2012
- James, Michelle L.; Gambhir, Sanjiv S.
- Physiological Reviews, Vol. 92, Issue 2
Extravasation of macromolecules
journal, October 1998
- Takakura, Yoshinobu; Mahato, Ram I.; Hashida, Mitsuru
- Advanced Drug Delivery Reviews, Vol. 34, Issue 1
The synthesis and characterization of novel coumarin dyes derived from 1,4-diethyl-1,2,3,4-tetrahydro-7-hydroxyquinoxalin-6-carboxaldehyde
journal, July 2009
- Jagtap, Amit R.; Satam, Vijay S.; Rajule, Rajkumar N.
- Dyes and Pigments, Vol. 82, Issue 1