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AZD-7648, a DNA-PK Inhibitor, Induces DNA Damage, Apoptosis, and Cell Cycle Arrest in Chronic and Acute Myeloid Leukemia Cells

The non-homologous end joining pathway is vital for repairing DNA double-strand breaks (DSB), with DNA-dependent protein kinase (DNA-PK) playing a critical role. Altered DNA damage response (DDR) in chronic (CML) and acute myeloid leukemia (AML) offers potential therapeutic opportunities. We studied...

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Autores principales: Lapa, Beatriz Santos, Costa, Maria Inês, Figueiredo, Diana, Jorge, Joana, Alves, Raquel, Monteiro, Ana Raquel, Serambeque, Beatriz, Laranjo, Mafalda, Botelho, Maria Filomena, Carreira, Isabel Marques, Sarmento-Ribeiro, Ana Bela, Gonçalves, Ana Cristina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10607085/
https://www.ncbi.nlm.nih.gov/pubmed/37895013
http://dx.doi.org/10.3390/ijms242015331
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author Lapa, Beatriz Santos
Costa, Maria Inês
Figueiredo, Diana
Jorge, Joana
Alves, Raquel
Monteiro, Ana Raquel
Serambeque, Beatriz
Laranjo, Mafalda
Botelho, Maria Filomena
Carreira, Isabel Marques
Sarmento-Ribeiro, Ana Bela
Gonçalves, Ana Cristina
author_facet Lapa, Beatriz Santos
Costa, Maria Inês
Figueiredo, Diana
Jorge, Joana
Alves, Raquel
Monteiro, Ana Raquel
Serambeque, Beatriz
Laranjo, Mafalda
Botelho, Maria Filomena
Carreira, Isabel Marques
Sarmento-Ribeiro, Ana Bela
Gonçalves, Ana Cristina
author_sort Lapa, Beatriz Santos
collection PubMed
description The non-homologous end joining pathway is vital for repairing DNA double-strand breaks (DSB), with DNA-dependent protein kinase (DNA-PK) playing a critical role. Altered DNA damage response (DDR) in chronic (CML) and acute myeloid leukemia (AML) offers potential therapeutic opportunities. We studied the therapeutic potential of AZD-7648 (DNA-PK inhibitor) in CML and AML cell lines. This study used two CML (K-562 and LAMA-84) and five AML (HEL, HL-60, KG-1, NB-4, and THP-1) cell lines. DDR gene mutations were obtained from the COSMIC database. The copy number and methylation profile were evaluated using MS-MLPA and DDR genes, and telomere length using qPCR. p53 protein expression was assessed using Western Blot, chromosomal damage through cytokinesis-block micronucleus assay, and γH2AX levels and DSB repair kinetics using flow cytometry. Cell density and viability were analyzed using trypan blue assay after treatment with AZD-7648 in concentrations ranging from 10 to 200 µM. Cell death, cell cycle distribution, and cell proliferation rate were assessed using flow cytometry. The cells displayed different DNA baseline damage, DDR gene expressions, mutations, genetic/epigenetic changes, and p53 expression. Only HEL cells displayed inefficient DSB repair. The LAMA-84, HEL, and KG-1 cells were the most sensitive to AZD-7648, whereas HL-60 and K-562 showed a lower effect on density and viability. Besides the reduction in cell proliferation, AZD-7648 induced apoptosis, cell cycle arrest, and DNA damage. In conclusion, these results suggest that AZD-7648 holds promise as a potential therapy for myeloid leukemias, however, with variations in drug sensitivity among tested cell lines, thus supporting further investigation to identify the specific factors influencing sensitivity to this DNA-PK inhibitor.
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spelling pubmed-106070852023-10-28 AZD-7648, a DNA-PK Inhibitor, Induces DNA Damage, Apoptosis, and Cell Cycle Arrest in Chronic and Acute Myeloid Leukemia Cells Lapa, Beatriz Santos Costa, Maria Inês Figueiredo, Diana Jorge, Joana Alves, Raquel Monteiro, Ana Raquel Serambeque, Beatriz Laranjo, Mafalda Botelho, Maria Filomena Carreira, Isabel Marques Sarmento-Ribeiro, Ana Bela Gonçalves, Ana Cristina Int J Mol Sci Article The non-homologous end joining pathway is vital for repairing DNA double-strand breaks (DSB), with DNA-dependent protein kinase (DNA-PK) playing a critical role. Altered DNA damage response (DDR) in chronic (CML) and acute myeloid leukemia (AML) offers potential therapeutic opportunities. We studied the therapeutic potential of AZD-7648 (DNA-PK inhibitor) in CML and AML cell lines. This study used two CML (K-562 and LAMA-84) and five AML (HEL, HL-60, KG-1, NB-4, and THP-1) cell lines. DDR gene mutations were obtained from the COSMIC database. The copy number and methylation profile were evaluated using MS-MLPA and DDR genes, and telomere length using qPCR. p53 protein expression was assessed using Western Blot, chromosomal damage through cytokinesis-block micronucleus assay, and γH2AX levels and DSB repair kinetics using flow cytometry. Cell density and viability were analyzed using trypan blue assay after treatment with AZD-7648 in concentrations ranging from 10 to 200 µM. Cell death, cell cycle distribution, and cell proliferation rate were assessed using flow cytometry. The cells displayed different DNA baseline damage, DDR gene expressions, mutations, genetic/epigenetic changes, and p53 expression. Only HEL cells displayed inefficient DSB repair. The LAMA-84, HEL, and KG-1 cells were the most sensitive to AZD-7648, whereas HL-60 and K-562 showed a lower effect on density and viability. Besides the reduction in cell proliferation, AZD-7648 induced apoptosis, cell cycle arrest, and DNA damage. In conclusion, these results suggest that AZD-7648 holds promise as a potential therapy for myeloid leukemias, however, with variations in drug sensitivity among tested cell lines, thus supporting further investigation to identify the specific factors influencing sensitivity to this DNA-PK inhibitor. MDPI 2023-10-18 /pmc/articles/PMC10607085/ /pubmed/37895013 http://dx.doi.org/10.3390/ijms242015331 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lapa, Beatriz Santos
Costa, Maria Inês
Figueiredo, Diana
Jorge, Joana
Alves, Raquel
Monteiro, Ana Raquel
Serambeque, Beatriz
Laranjo, Mafalda
Botelho, Maria Filomena
Carreira, Isabel Marques
Sarmento-Ribeiro, Ana Bela
Gonçalves, Ana Cristina
AZD-7648, a DNA-PK Inhibitor, Induces DNA Damage, Apoptosis, and Cell Cycle Arrest in Chronic and Acute Myeloid Leukemia Cells
title AZD-7648, a DNA-PK Inhibitor, Induces DNA Damage, Apoptosis, and Cell Cycle Arrest in Chronic and Acute Myeloid Leukemia Cells
title_full AZD-7648, a DNA-PK Inhibitor, Induces DNA Damage, Apoptosis, and Cell Cycle Arrest in Chronic and Acute Myeloid Leukemia Cells
title_fullStr AZD-7648, a DNA-PK Inhibitor, Induces DNA Damage, Apoptosis, and Cell Cycle Arrest in Chronic and Acute Myeloid Leukemia Cells
title_full_unstemmed AZD-7648, a DNA-PK Inhibitor, Induces DNA Damage, Apoptosis, and Cell Cycle Arrest in Chronic and Acute Myeloid Leukemia Cells
title_short AZD-7648, a DNA-PK Inhibitor, Induces DNA Damage, Apoptosis, and Cell Cycle Arrest in Chronic and Acute Myeloid Leukemia Cells
title_sort azd-7648, a dna-pk inhibitor, induces dna damage, apoptosis, and cell cycle arrest in chronic and acute myeloid leukemia cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10607085/
https://www.ncbi.nlm.nih.gov/pubmed/37895013
http://dx.doi.org/10.3390/ijms242015331
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