Identification of proteins and cellular pathways targeted by 2-nitroimidazole hypoxic cytotoxins

Rashed, Faisal Bin; Stoica, Alexandru Cezar; Macdonald, Dawn; Elsaidi, Hassan R. H.; Ricardo, Carolynne; Bhatt, Bhumi; Moore, Jack; Diaz‐Dussan, Diana; Ramamonjisoa, Nirilanto; Mowery, Yvonne M.; Damaraju, Sambasivarao; Fahlman, Richard P.; Kumar, Piyush; Weinfeld, Michael

doi:10.1016/j.redox.2021.101905

Cited by 6 publications

(27 citation statements)

References 60 publications

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“…Synthesis of IAZA [ 12 ], FAZA [ 13 ] and azidoazomycin arabinofuranoside (N 3 -AZA) [ 20 ] have been described previously. IAZA, FAZA, N 3 -AZA and Ferrostatin-1 (SML0583, Sigma-Aldrich, Oakville, ON) were prepared as 500 mM solutions in DMSO and stored at −20 °C.…”

Section: Methodsmentioning

confidence: 99%

“…Instead of antibody based detection, our method utilizes click chemistry by using a click chemistry compatible 2-NI compound, N 3 -AZA ( Fig. 1 C) [ 20 ]. Briefly, FaDu cells treated with 100 μM N 3 -AZA (or 0.02% DMSO) under normoxia or hypoxia (<0.1% O 2 ) for 24 h were harvested by trypsinization and pelleted by centrifugation.…”

Section: Methodsmentioning

confidence: 99%

“…FaDu cells treated with IAZA (150 μM), FAZA (350 μM) or vehicle control (0.02% DMSO) for 24 h under normoxia or hypoxia (<0.1% O 2 ) were harvested by trypsinization and pelleted by centrifugation. Cellular GAPDH enzyme activity was quantified as described earlier using a GAPDH activity assay kit (ab204732, Abcam) [ 20 ].…”

Section: Methodsmentioning

confidence: 99%

“…Total cellular GST enzymatic activity was measured using a GST assay kit (CS0410, Sigma-Aldrich). FaDu cells treated with IAZA (100 μM and 150 μM), FAZA (100 μM and 350 μM) or vehicle control (0.02% DMSO) for 24 h under normoxia or hypoxia (<0.1% O 2 ) were processed to quantify total GST activity as previously described [ 20 ].…”

Section: Methodsmentioning

confidence: 99%

“…with N 3 -AZA (60 mg/kg b.w.). After euthanization, tumours were excised, flash frozen in optimal cutting temperature compound (O.C.T., 4585, Fisher Scientific, Waltham, MA), and N 3 -AZA click chemistry was performed on central sagittal tumour sections to determine endpoint hypoxia levels as described previously [ 20 ]. Briefly, acetone-fixed frozen tumour sections were blocked in PBS with 1% BSA, incubated with primary antibody for small vessel endothelium (rat anti-CD31, 1:50 dilution; 550274, BD Pharmingen) for 2 h, followed by incubation with the secondary antibody (Alexa Fluor 647 goat anti-rat IgG, 1:1000 dilution; Molecular Probes) mixed with 4′,6-diamidino-2-phenylindole (DAPI).…”

Section: Methodsmentioning

confidence: 99%

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Cellular mechanism of action of 2-nitroimidazoles as hypoxia-selective therapeutic agents

et al. 2022

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Cellular mechanism of action of 2-nitroimidazoles as hypoxia-selective therapeutic agents

et al. 2022

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Pimonidazole-alkyne conjugate for sensitive detection of hypoxia by Cu-catalyzed click reaction

Tamura,

Sakamoto,

et al. 2024

ANAL. SCI.

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Hypoxia is involved in various diseases, such as cancers. Pimonidazole has often been used as the gold-standard marker to visualize hypoxic regions. Pimonidazole labels hypoxic regions by forming a covalent bond with a neighboring protein under hypoxic conditions in the body, which is detected by immunohistochemistry performed on tissue sections. To date, some pimonidazole-fluorophore conjugates have been reported as fluorescent probes for hypoxia imaging that do not require immunostaining. They are superior to pimonidazole because immunostaining can produce high background signals. However, large fluorophores in the conjugates may alter the original biodistribution and reactivity. Here, we report a new hypoxia marker, Pimo-yne, as a pimonidazole-alkyne conjugate. Pimo-yne has a similar hypoxia detection capability as pimonidazole because the alkyne tag is small and can be detected by Cu-catalyzed click reaction with azide-tagged fluorescent dyes. We successfully visualized hypoxic regions in tumor tissue sections using Pimo-yne with reduced background signals. The detected regions overlapped well with those detected by pimonidazole immunohistochemistry. To further reduce the background, we employed a turn-on azide-tagged fluorescent dye. Graphical abstract

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Optimized Carbohydrate-Based Nanogel Formulation to Sensitize Hypoxic Tumors

et al. 2023

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Solid tumors are often poorly vascularized, which impairs oxygen supply and drug delivery to the cells. This often leads to genetic and translational adaptations that promote tumor progression, invasion, metastasis, and resistance to conventional chemo-/radiotherapy and immunotherapy. A hypoxia-directed nanosensitizer formulation of a hypoxia-activated prodrug (HAP) was developed by encapsulating iodoazomycin arabinofuranoside (IAZA), a 2-nitroimidazole nucleoside-based HAP, in a functionally modified carbohydrate-based nanogel, facilitating delivery and accrual selectively in the hypoxic head and neck and prostate cancer cells. Although IAZA has been reported as a clinically validated hypoxia diagnostic agent, recent studies have pointed to its promising hypoxia-selective anti-tumor properties, which make IAZA an excellent candidate for further exploration as a multimodal theranostic of hypoxic tumors. The nanogels are composed of a galactose-based shell with an inner core of thermoresponsive (di(ethylene glycol) methyl ethyl methacrylate) (DEGMA). Optimization of the nanogels led to high IAZA-loading capacity (≅80–88%) and a slow time-controlled release over 50 h. Furthermore, nanoIAZA (encapsulated IAZA) displayed superior in vitro hypoxia-selective cytotoxicity and radiosensitization in comparison to free IAZA in the head and neck (FaDu) and prostate (PC3) cancer cell lines. The acute systemic toxicity profile of the nanogel (NG1) was studied in immunocompromised mice, indicating no signs of toxicity. Additionally, growth inhibition of subcutaneous FaDu xenograft tumors was observed with nanoIAZA, demonstrating that this nanoformulation offers a significant improvement in tumor regression and overall survival compared to the control.

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Identification of proteins and cellular pathways targeted by 2-nitroimidazole hypoxic cytotoxins

Cited by 6 publications

References 60 publications

Cellular mechanism of action of 2-nitroimidazoles as hypoxia-selective therapeutic agents

Cellular mechanism of action of 2-nitroimidazoles as hypoxia-selective therapeutic agents

Pimonidazole-alkyne conjugate for sensitive detection of hypoxia by Cu-catalyzed click reaction

Optimized Carbohydrate-Based Nanogel Formulation to Sensitize Hypoxic Tumors

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