Detecting Attomolar DNA-Damaging Anticancer Drug Activity in Cell Lysates with Electrochemical DNA Devices
| Autor: | Reema McMullen, Colton L. Starcher, Edward A. Motea, Zakari Ishak-Boushaki, Chloe C. DiTusa, Dimithree Kahanda, Naveen Singh, Jason D. Slinker, Ashan P. Wettasinghe |
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| Rok vydání: | 2021 |
| Předmět: |
Drug
media_common.quotation_subject Bioengineering Antineoplastic Agents 02 engineering and technology 01 natural sciences Article chemistry.chemical_compound Western blot Cell Line Tumor medicine NAD(P)H Dehydrogenase (Quinone) Potency Instrumentation media_common Fluid Flow and Transfer Processes medicine.diagnostic_test Process Chemistry and Technology 010401 analytical chemistry Base excision repair DNA 021001 nanoscience & nanotechnology Orders of magnitude (mass) 0104 chemical sciences Kinetics Biochemistry chemistry Cancer cell NAD+ kinase 0210 nano-technology Naphthoquinones |
| Zdroj: | ACS Sens |
| ISSN: | 2379-3694 |
| Popis: | Here, we utilize electrochemical DNA devices to quantify and understand the cancer-specific DNA-damaging activity of an emerging drug in cellular lysates at femtomolar and attomolar concentrations. Isobutyl-deoxynyboquinone (IB-DNQ), a potent and tumor-selective NAD(P)H quinone oxidoreductase 1 (NQO1) bioactivatable drug, was prepared and biochemically verified in cancer cells highly expressing NQO1 (NQO1+) and knockdowns with low NQO1 expression (NQO1-) by Western blot, NQO1 activity analysis, survival assays, oxygen consumption rate, extracellular acidification rate, and peroxide production. Lysates from these cells and the IB-DNQ drug were then introduced to a chip system bearing an array of DNA-modified electrodes, and their DNA-damaging activity was quantified by changes in DNA-mediated electrochemistry arising from base-excision repair. Device-level controls of NQO1 activity and kinetic analysis were used to verify and further understand the IB-DNQ activity. A 380 aM IB-DNQ limit of detection and a 1.3 fM midpoint of damage were observed in NQO1+ lysates, both metrics 2 orders of magnitude lower than NQO1- lysates, indicating the high IB-DNQ potency and selectivity for NQO1+ cancers. The device-level damage midpoint concentration in NQO1+ lysates was over 8 orders of magnitude lower than cell survival benchmarks, likely due to poor IB-DNQ cellular uptake, demonstrating that these devices can identify promising drugs requiring improved cell permeability. Ultimately, these results indicate the noteworthy potency and selectivity of IB-DNQ and the high sensitivity and precision of electrochemical DNA devices to analyze agents/drugs involved in DNA-damaging chemotherapies. |
| Databáze: | OpenAIRE |
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