In vitro activity of a G-quadruplex-stabilizing small molecule that synergizes with Navitoclax to induce cytotoxicity in acute myeloid leukemia cells
Abstract Background Acute Myeloid Leukemia (AML) is a malignancy of myeloid precursor cells that arise from genomic alterations in the expression of key growth regulatory genes causing cells to assume an undifferentiated state and continue to proliferate. Recent efforts have focused on developing th...
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| Format: | Article |
| Language: | English |
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BMC
2019-12-01
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| Series: | BMC Cancer |
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| Online Access: | https://doi.org/10.1186/s12885-019-6464-9 |
| _version_ | 1818386986620157952 |
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| author | Justin J. Montoya Megan A. Turnidge Daniel H. Wai Apurvi R. Patel David W. Lee Vijay Gokhale Laurence H. Hurley Robert J. Arceci Cynthia Wetmore David O. Azorsa |
| author_facet | Justin J. Montoya Megan A. Turnidge Daniel H. Wai Apurvi R. Patel David W. Lee Vijay Gokhale Laurence H. Hurley Robert J. Arceci Cynthia Wetmore David O. Azorsa |
| author_sort | Justin J. Montoya |
| collection | DOAJ |
| description | Abstract Background Acute Myeloid Leukemia (AML) is a malignancy of myeloid precursor cells that arise from genomic alterations in the expression of key growth regulatory genes causing cells to assume an undifferentiated state and continue to proliferate. Recent efforts have focused on developing therapies that target specific protein products of aberrantly expressed genes. However, many of the identified proteins are difficult to target and thought to be “undrugable” because of structural challenges, protein overexpression, or mutations that confer resistance to therapy. A novel technology that circumvents some of these issues is the use of small molecules that stabilize secondary DNA structures present in the promoters of many potential oncogenes and modulate their transcription. Methods This study characterizes the in vitro activity of the G-quadruplex-stabilizing small molecule GQC-05 in AML cells. The effect of GQC-05 on three AML cell lines was analyzed using viability and apoptosis assays. GQC-05 has been shown to down-regulate MYC through G-quadruplex stabilization in Burkitt’s lymphoma cell lines. MYC expression was evaluated through qPCR and immunoblotting in the three AML cell lines following the treatment of GQC-05. In order to identify other therapeutic agents that potentiate the activity of GQC-05, combination drug screening was performed. The drug combinations were validated using in vitro cytotoxicity assays and compared to other commonly used chemotherapeutic agents. Results GQC-05 treatment of KG-1a, CMK and TF-1 cells decreased cell viability and resulted in increased DNA damage and apoptosis. Additionally, treatment of KG-1a, CMK and TF-1 with GQC-05 resulted in decreased expression of MYC mRNA and protein, with a more pronounced effect in KG-1a cells. Combination drug screening identified the Bcl-2/Bcl-XL inhibitor Navitoclax as a compound that potentiated GQC-05 activity. Co-treatment with GQC-05 and Navitoclax showed a synergistic decrease in cell viability of AML cells as determined by Chou-Talalay analysis, and induced more DNA damage, apoptosis, and rapid cytotoxicity. The cytotoxicity induced by GQC-05 and Navitoclax was more potent than that of Navitoclax combined with either cytarabine or doxorubicin. Conclusion These results suggest that the G-quadruplex stabilizing small molecule GQC-05 induces down regulated MYC expression and DNA damage in AML cells. Treatment with both GQC-05 with a Bcl-2/Bcl-XL inhibitor Navitoclax results in increased cytotoxic activity, which is more pronounced than Navitoclax or GQC-05 alone, and more significant than Navitoclax in combination with cytarabine and doxorubicin that are currently being used clinically. |
| first_indexed | 2024-12-14T04:02:46Z |
| format | Article |
| id | doaj.art-70b17a3e059a4dbc847f00fba0fb5ba6 |
| institution | Directory Open Access Journal |
| issn | 1471-2407 |
| language | English |
| last_indexed | 2024-12-14T04:02:46Z |
| publishDate | 2019-12-01 |
| publisher | BMC |
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| series | BMC Cancer |
| spelling | doaj.art-70b17a3e059a4dbc847f00fba0fb5ba62022-12-21T23:17:54ZengBMCBMC Cancer1471-24072019-12-0119111110.1186/s12885-019-6464-9In vitro activity of a G-quadruplex-stabilizing small molecule that synergizes with Navitoclax to induce cytotoxicity in acute myeloid leukemia cellsJustin J. Montoya0Megan A. Turnidge1Daniel H. Wai2Apurvi R. Patel3David W. Lee4Vijay Gokhale5Laurence H. Hurley6Robert J. Arceci7Cynthia Wetmore8David O. Azorsa9The Institute of Molecular Medicine at Phoenix Children’s HospitalThe Institute of Molecular Medicine at Phoenix Children’s HospitalThe Institute of Molecular Medicine at Phoenix Children’s HospitalThe Institute of Molecular Medicine at Phoenix Children’s HospitalThe Institute of Molecular Medicine at Phoenix Children’s HospitalUniversity of Arizona College of PharmacyUniversity of Arizona College of PharmacyThe Institute of Molecular Medicine at Phoenix Children’s HospitalThe Institute of Molecular Medicine at Phoenix Children’s HospitalThe Institute of Molecular Medicine at Phoenix Children’s HospitalAbstract Background Acute Myeloid Leukemia (AML) is a malignancy of myeloid precursor cells that arise from genomic alterations in the expression of key growth regulatory genes causing cells to assume an undifferentiated state and continue to proliferate. Recent efforts have focused on developing therapies that target specific protein products of aberrantly expressed genes. However, many of the identified proteins are difficult to target and thought to be “undrugable” because of structural challenges, protein overexpression, or mutations that confer resistance to therapy. A novel technology that circumvents some of these issues is the use of small molecules that stabilize secondary DNA structures present in the promoters of many potential oncogenes and modulate their transcription. Methods This study characterizes the in vitro activity of the G-quadruplex-stabilizing small molecule GQC-05 in AML cells. The effect of GQC-05 on three AML cell lines was analyzed using viability and apoptosis assays. GQC-05 has been shown to down-regulate MYC through G-quadruplex stabilization in Burkitt’s lymphoma cell lines. MYC expression was evaluated through qPCR and immunoblotting in the three AML cell lines following the treatment of GQC-05. In order to identify other therapeutic agents that potentiate the activity of GQC-05, combination drug screening was performed. The drug combinations were validated using in vitro cytotoxicity assays and compared to other commonly used chemotherapeutic agents. Results GQC-05 treatment of KG-1a, CMK and TF-1 cells decreased cell viability and resulted in increased DNA damage and apoptosis. Additionally, treatment of KG-1a, CMK and TF-1 with GQC-05 resulted in decreased expression of MYC mRNA and protein, with a more pronounced effect in KG-1a cells. Combination drug screening identified the Bcl-2/Bcl-XL inhibitor Navitoclax as a compound that potentiated GQC-05 activity. Co-treatment with GQC-05 and Navitoclax showed a synergistic decrease in cell viability of AML cells as determined by Chou-Talalay analysis, and induced more DNA damage, apoptosis, and rapid cytotoxicity. The cytotoxicity induced by GQC-05 and Navitoclax was more potent than that of Navitoclax combined with either cytarabine or doxorubicin. Conclusion These results suggest that the G-quadruplex stabilizing small molecule GQC-05 induces down regulated MYC expression and DNA damage in AML cells. Treatment with both GQC-05 with a Bcl-2/Bcl-XL inhibitor Navitoclax results in increased cytotoxic activity, which is more pronounced than Navitoclax or GQC-05 alone, and more significant than Navitoclax in combination with cytarabine and doxorubicin that are currently being used clinically.https://doi.org/10.1186/s12885-019-6464-9MYCG-quadruplexAMLBcl-2Bcl-XLNavitoclax |
| spellingShingle | Justin J. Montoya Megan A. Turnidge Daniel H. Wai Apurvi R. Patel David W. Lee Vijay Gokhale Laurence H. Hurley Robert J. Arceci Cynthia Wetmore David O. Azorsa In vitro activity of a G-quadruplex-stabilizing small molecule that synergizes with Navitoclax to induce cytotoxicity in acute myeloid leukemia cells BMC Cancer MYC G-quadruplex AML Bcl-2 Bcl-XL Navitoclax |
| title | In vitro activity of a G-quadruplex-stabilizing small molecule that synergizes with Navitoclax to induce cytotoxicity in acute myeloid leukemia cells |
| title_full | In vitro activity of a G-quadruplex-stabilizing small molecule that synergizes with Navitoclax to induce cytotoxicity in acute myeloid leukemia cells |
| title_fullStr | In vitro activity of a G-quadruplex-stabilizing small molecule that synergizes with Navitoclax to induce cytotoxicity in acute myeloid leukemia cells |
| title_full_unstemmed | In vitro activity of a G-quadruplex-stabilizing small molecule that synergizes with Navitoclax to induce cytotoxicity in acute myeloid leukemia cells |
| title_short | In vitro activity of a G-quadruplex-stabilizing small molecule that synergizes with Navitoclax to induce cytotoxicity in acute myeloid leukemia cells |
| title_sort | in vitro activity of a g quadruplex stabilizing small molecule that synergizes with navitoclax to induce cytotoxicity in acute myeloid leukemia cells |
| topic | MYC G-quadruplex AML Bcl-2 Bcl-XL Navitoclax |
| url | https://doi.org/10.1186/s12885-019-6464-9 |
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