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Repurposing of the ALK Inhibitor Crizotinib for Acute Leukemia and Multiple Myeloma Cells

Crizotinib was a first generation of ALK tyrosine kinase inhibitor approved for the treatment of ALK-positive non-small-cell lung carcinoma (NSCLC) patients. COMPARE and cluster analyses of transcriptomic data of the NCI cell line panel indicated that genes with different cellular functions regulate...

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Autores principales: Boulos, Joelle C., Saeed, Mohamed E. M., Chatterjee, Manik, Bülbül, Yagmur, Crudo, Francesco, Marko, Doris, Munder, Markus, Klauck, Sabine M., Efferth, Thomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8617756/
https://www.ncbi.nlm.nih.gov/pubmed/34832908
http://dx.doi.org/10.3390/ph14111126
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author Boulos, Joelle C.
Saeed, Mohamed E. M.
Chatterjee, Manik
Bülbül, Yagmur
Crudo, Francesco
Marko, Doris
Munder, Markus
Klauck, Sabine M.
Efferth, Thomas
author_facet Boulos, Joelle C.
Saeed, Mohamed E. M.
Chatterjee, Manik
Bülbül, Yagmur
Crudo, Francesco
Marko, Doris
Munder, Markus
Klauck, Sabine M.
Efferth, Thomas
author_sort Boulos, Joelle C.
collection PubMed
description Crizotinib was a first generation of ALK tyrosine kinase inhibitor approved for the treatment of ALK-positive non-small-cell lung carcinoma (NSCLC) patients. COMPARE and cluster analyses of transcriptomic data of the NCI cell line panel indicated that genes with different cellular functions regulated the sensitivity or resistance of cancer cells to crizotinib. Transcription factor binding motif analyses in gene promoters divulged two transcription factors possibly regulating the expression of these genes, i.e., RXRA and GATA1, which are important for leukemia and erythroid development, respectively. COMPARE analyses also implied that cell lines of various cancer types displayed varying degrees of sensitivity to crizotinib. Unexpectedly, leukemia but not lung cancer cells were the most sensitive cells among the different types of NCI cancer cell lines. Re-examining this result in another panel of cell lines indeed revealed that crizotinib exhibited potent cytotoxicity towards acute myeloid leukemia and multiple myeloma cells. P-glycoprotein-overexpressing CEM/ADR5000 leukemia cells were cross-resistant to crizotinib. NCI-H929 multiple myeloma cells were the most sensitive cells. Hence, we evaluated the mode of action of crizotinib on these cells. Although crizotinib is a TKI, it showed highest correlation rates with DNA topoisomerase II inhibitors and tubulin inhibitors. The altered gene expression profiles after crizotinib treatment predicted several networks, where TOP2A and genes related to cell cycle were downregulated. Cell cycle analyses showed that cells incubated with crizotinib for 24 h accumulated in the G(2)M phase. Crizotinib also increased the number of p-H3(Ser10)-positive NCI-H929 cells illustrating crizotinib’s ability to prevent mitotic exit. However, cells accumulated in the sub-G(0)G(1) fraction with longer incubation periods, indicating apoptosis induction. Additionally, crizotinib disassembled the tubulin network of U2OS cells expressing an α-tubulin-GFP fusion protein, preventing migration of cancer cells. This result was verified by in vitro tubulin polymerization assays. In silico molecular docking also revealed a strong binding affinity of crizotinib to the colchicine and Vinca alkaloid binding sites. Taken together, these results demonstrate that crizotinib destabilized microtubules. Additionally, the decatenation assay showed that crizotinib partwise inhibited the catalytic activity of DNA topoisomerase II. In conclusion, crizotinib exerted kinase-independent cytotoxic effects through the dual inhibition of tubulin polymerization and topoisomerase II and might be used to treat not only NSCLC but also multiple myeloma.
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spelling pubmed-86177562021-11-27 Repurposing of the ALK Inhibitor Crizotinib for Acute Leukemia and Multiple Myeloma Cells Boulos, Joelle C. Saeed, Mohamed E. M. Chatterjee, Manik Bülbül, Yagmur Crudo, Francesco Marko, Doris Munder, Markus Klauck, Sabine M. Efferth, Thomas Pharmaceuticals (Basel) Article Crizotinib was a first generation of ALK tyrosine kinase inhibitor approved for the treatment of ALK-positive non-small-cell lung carcinoma (NSCLC) patients. COMPARE and cluster analyses of transcriptomic data of the NCI cell line panel indicated that genes with different cellular functions regulated the sensitivity or resistance of cancer cells to crizotinib. Transcription factor binding motif analyses in gene promoters divulged two transcription factors possibly regulating the expression of these genes, i.e., RXRA and GATA1, which are important for leukemia and erythroid development, respectively. COMPARE analyses also implied that cell lines of various cancer types displayed varying degrees of sensitivity to crizotinib. Unexpectedly, leukemia but not lung cancer cells were the most sensitive cells among the different types of NCI cancer cell lines. Re-examining this result in another panel of cell lines indeed revealed that crizotinib exhibited potent cytotoxicity towards acute myeloid leukemia and multiple myeloma cells. P-glycoprotein-overexpressing CEM/ADR5000 leukemia cells were cross-resistant to crizotinib. NCI-H929 multiple myeloma cells were the most sensitive cells. Hence, we evaluated the mode of action of crizotinib on these cells. Although crizotinib is a TKI, it showed highest correlation rates with DNA topoisomerase II inhibitors and tubulin inhibitors. The altered gene expression profiles after crizotinib treatment predicted several networks, where TOP2A and genes related to cell cycle were downregulated. Cell cycle analyses showed that cells incubated with crizotinib for 24 h accumulated in the G(2)M phase. Crizotinib also increased the number of p-H3(Ser10)-positive NCI-H929 cells illustrating crizotinib’s ability to prevent mitotic exit. However, cells accumulated in the sub-G(0)G(1) fraction with longer incubation periods, indicating apoptosis induction. Additionally, crizotinib disassembled the tubulin network of U2OS cells expressing an α-tubulin-GFP fusion protein, preventing migration of cancer cells. This result was verified by in vitro tubulin polymerization assays. In silico molecular docking also revealed a strong binding affinity of crizotinib to the colchicine and Vinca alkaloid binding sites. Taken together, these results demonstrate that crizotinib destabilized microtubules. Additionally, the decatenation assay showed that crizotinib partwise inhibited the catalytic activity of DNA topoisomerase II. In conclusion, crizotinib exerted kinase-independent cytotoxic effects through the dual inhibition of tubulin polymerization and topoisomerase II and might be used to treat not only NSCLC but also multiple myeloma. MDPI 2021-11-05 /pmc/articles/PMC8617756/ /pubmed/34832908 http://dx.doi.org/10.3390/ph14111126 Text en © 2021 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
Boulos, Joelle C.
Saeed, Mohamed E. M.
Chatterjee, Manik
Bülbül, Yagmur
Crudo, Francesco
Marko, Doris
Munder, Markus
Klauck, Sabine M.
Efferth, Thomas
Repurposing of the ALK Inhibitor Crizotinib for Acute Leukemia and Multiple Myeloma Cells
title Repurposing of the ALK Inhibitor Crizotinib for Acute Leukemia and Multiple Myeloma Cells
title_full Repurposing of the ALK Inhibitor Crizotinib for Acute Leukemia and Multiple Myeloma Cells
title_fullStr Repurposing of the ALK Inhibitor Crizotinib for Acute Leukemia and Multiple Myeloma Cells
title_full_unstemmed Repurposing of the ALK Inhibitor Crizotinib for Acute Leukemia and Multiple Myeloma Cells
title_short Repurposing of the ALK Inhibitor Crizotinib for Acute Leukemia and Multiple Myeloma Cells
title_sort repurposing of the alk inhibitor crizotinib for acute leukemia and multiple myeloma cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8617756/
https://www.ncbi.nlm.nih.gov/pubmed/34832908
http://dx.doi.org/10.3390/ph14111126
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