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Combinatorial targeting of telomerase and DNA-PK induces synergistic apoptotic effects against Pre-B acute lymphoblastic leukemia cells

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Abstract

Background

Due to the high demand for novel approaches for leukemia-targeted therapy, this study investigates the impact of DNA-PK inhibitor NU7441 on the sensitivity of pre-B ALL cells to the telomerase inhibitor MST-312.

Methods

The study involved NALM-6 cells treated with MST-312 and NU7441, assessing their viability and metabolic activity using trypan blue and MTT assays. The study also evaluated apoptosis, gene expression changes, and DNA damage using flow cytometry, qRT-PCR, and micronucleus assays. The binding energy of MST-312 in the active site of telomerase was calculated using molecular docking.

Results

The study’s findings revealed a synergistic decline in both cell viability and metabolic activity in NALM-6 cells when exposed to the combined treatment of MST-312 and NU7441, and this decrease occurred without any adverse effects on healthy PBMC cells. Furthermore, the combination treatment exhibited a significantly higher induction of apoptosis than treatment with MST-312 alone, as observed through flow cytometry assay. qRT-PCR analysis revealed that this enhanced apoptosis was associated with a notable downregulation of Bcl-2 expression and an upregulation of Bax gene expression. Moreover, the combination therapy decreased expression levels of hTERT and c-Myc genes. The micronucleus assay indicated that the combination treatment increased DNA damage in NALM-6 cells. Also, a good conformation between MST-312 and the active site of telomerase was revealed by docking data.

Conclusions

The study suggests that simultaneous inhibition of telomerase and DNA-PK in pre-B ALL presents a novel targeted therapy approach.

Highlights

MST-312/NU7441 exerts a synergistic cytotoxic effect against NALM-6 cells.

NU7441 accentuates MST-312-induced apoptosis of NALM-6 cells.

MST-312/NU7441-induced apoptosis progresses through upregulation of the Bax/Bcl-2 ratio.

A combination of MST-312/NU7441 downregulates c-Myc and hTERT gene expression.

NU7441 increases MST-312-induced DNA damage in NALM-6 cells.

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Data Availability

The data would be available from the corresponding author upon reasonable request.

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Acknowledgements

The authors are willing to express their sincere thanks for the support provided by the Kerman University of Medical Sciences for this project.

Funding

This study was supported by grant No. 97000840 from the Kerman University of Medical Sciences.

Author information

Authors and Affiliations

  1. Student Research Committee, Faculty of Allied Medicine, Kerman University of Medical Sciences, Kerman, Iran

    Maryam Katoueezadeh & Parisa Maleki

  2. Department of Hematology and Medical Laboratory Sciences, Faculty of Allied Medicine, Kerman University of Medical Sciences, Kerman, Iran

    Maryam Katoueezadeh, Parisa Maleki, Alireza Farsinejad, Roohollah Mirzaee Khalilabadi, Hajar Mardani Valandani & Muhammad Hossein Ashoub

  3. Pharmaceutics Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran

    Seyedeh Atekeh Torabizadeh

  4. Cell Therapy and Regenerative Medicine Comprehensive Center, Kerman University of Medical Sciences, Kerman, Iran

    Alireza Farsinejad & Muhammad Hossein Ashoub

  5. Postdoctoral Research Fellow, Faculty of Applied Health Sciences, Brock University, St. Catharines, ON, Canada

    Ismaila Olanrewaju Nurain

  6. Cellular and Molecular Research Center, Gerash University of Medical Sciences, Gerash, Iran

    Ahmad Fatemi

Authors
  1. Maryam Katoueezadeh
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  2. Parisa Maleki
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  3. Seyedeh Atekeh Torabizadeh
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  4. Alireza Farsinejad
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  7. Ismaila Olanrewaju Nurain
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  8. Muhammad Hossein Ashoub
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Contributions

Maryam Katoueezadeh: Investigation, Writing- Original draft preparation, Visualization, Methodology. Parisa Maleki: Methodology, Writing- Reviewing and Editing. Seyedeh Atekeh Torabizadeh: Methodology, Writing- Reviewing and Editing. Alireza Farsinejad: Resources, Formal analysis, Writing- Reviewing and Editing. Roohollah Mirzaee Khalilabadi: Methodology, Writing- Reviewing and Editing. Hajar Mardani Valandani: Methodology, Writing- Reviewing and Editing. Ismaila Olanrewaju Nurain: Methodology, Writing- Reviewing and Editing. Muhammad Hossein Ashoub: Formal analysis, Methodology, Writing- Reviewing and Editing. Ahmad Fatemi: Conceptualization, Supervision, Methodology, Validation, Writing- Reviewing and Editing.

Corresponding author

Correspondence to Ahmad Fatemi.

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The study gained the approval of the ethics committee of the Kerman University of Medical Sciences with the ethical approval code IR.KMU.REC.1397.589. All methods were performed following the relevant guidelines and regulations.

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Katoueezadeh, M., Maleki, P., Torabizadeh, S.A. et al. Combinatorial targeting of telomerase and DNA-PK induces synergistic apoptotic effects against Pre-B acute lymphoblastic leukemia cells. Mol Biol Rep 51, 163 (2024). https://doi.org/10.1007/s11033-023-09087-9

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