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      The Aurora kinase B relocation blocker LXY18 triggers mitotic catastrophe selectively in malignant cells

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          Abstract

          The mitotic regulator, Aurora kinase B (AURKB), is frequently overexpressed in malignancy and is a target for therapeutic intervention. The compound, LXY18, is a potent, orally available small molecule that inhibits the proper localization of AURKB during late mitosis, without affecting its kinase activity. In this study, we demonstrate that LXY18 elicits apoptosis in cancer cells derived from various indications, but not in non-transformed cell lines. The apoptosis is p53-independent, triggered by a prolonged mitotic arrest and occurs predominantly in mitosis. Some additional cells succumb post-mitotic slippage. We also demonstrate that cancer cell lines refractory to AURKB kinase inhibitors are sensitive to LXY18. The mitotic proteins MKLP2, NEK6, NEK7 and NEK9 are known regulators of AURKB localization during the onset of anaphase. LXY18 fails to inhibit the catalytic activity of these AURKB localization factors. Overall, our findings suggest a novel activity for LXY18 that produces a prolonged mitotic arrest and lethality in cancer cells, leaving non-transformed cells healthy. This new activity suggests that the compound may be a promising drug candidate for cancer treatment and that it can also be used as a tool compound to further dissect the regulatory network controlling AURKB localization.

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          Most cited references37

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          Chromosomal passengers: conducting cell division.

          Mitosis and meiosis are remarkable processes during which cells undergo profound changes in their structure and physiology. These events are orchestrated with a precision that is worthy of a classical symphony, with different activities being switched on and off at precise times and locations throughout the cell. One essential 'conductor' of this symphony is the chromosomal passenger complex (CPC), which comprises Aurora-B protein kinase, the inner centromere protein INCENP, survivin and borealin (also known as Dasra-B). Studies of the CPC are providing insights into its functions, which range from chromosome-microtubule interactions to sister chromatid cohesion to cytokinesis, and constitute one of the most dynamic areas of ongoing mitosis and meiosis research.
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            The chromosomal passenger complex (CPC): from easy rider to the godfather of mitosis.

            Successful cell division requires the precise and timely coordination of chromosomal, cytoskeletal and membrane trafficking events. These processes are regulated by the competing actions of protein kinases and phosphatases. Aurora B is one of the most intensively studied kinases. In conjunction with inner centromere protein (INCENP), borealin (also known as Dasra) and survivin it forms the chromosomal passenger complex (CPC). This complex targets to different locations at differing times during mitosis, where it regulates key mitotic events: correction of chromosome-microtubule attachment errors; activation of the spindle assembly checkpoint; and construction and regulation of the contractile apparatus that drives cytokinesis. Our growing understanding of the CPC has seen it develop from a mere passenger riding on the chromosomes to one of the main controllers of mitosis.
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              Exploring the functional interactions between Aurora B, INCENP, and survivin in mitosis.

              The function of the Aurora B kinase at centromeres and the central spindle is crucial for chromosome segregation and cytokinesis, respectively. Herein, we have investigated the regulation of human Aurora B by its complex partners inner centromere protein (INCENP) and survivin. We found that overexpression of a catalytically inactive, dominant-negative mutant of Aurora B impaired the localization of the entire Aurora B/INCENP/survivin complex to centromeres and the central spindle and severely disturbed mitotic progression. Similar results were also observed after depletion, by RNA interference, of either Aurora B, INCENP, or survivin. These data suggest that Aurora B kinase activity and the formation of the Aurora B/INCENP/survivin complex both contribute to its proper localization. Using recombinant proteins, we found that Aurora B kinase activity was stimulated by INCENP and that the C-terminal region of INCENP was sufficient for activation. Under identical assay conditions, survivin did not detectably influence kinase activity. Human INCENP was a substrate of Aurora B and mass spectrometry identified three consecutive residues (threonine 893, serine 894, and serine 895) containing at least two phosphorylation sites. A nonphosphorylatable mutant (TSS893-895AAA) was a poor activator of Aurora B, demonstrating that INCENP phosphorylation is important for kinase activation.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: MethodologyRole: Writing – original draft
                Role: Data curationRole: MethodologyRole: Writing – original draft
                Role: Data curation
                Role: Data curation
                Role: Data curation
                Role: ConceptualizationRole: Data curationRole: Methodology
                Role: Data curation
                Role: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS One
                plos
                PLOS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                30 October 2023
                2023
                : 18
                : 10
                : e0293283
                Affiliations
                [1 ] Division of Discovery Oncology, Chengdu Anticancer Bioscience, Chengdu, Sichuan, China
                [2 ] Department of Basic Cancer Research, J. Michael Bishop Institute of Cancer Research, Chengdu, Sichuan, China
                Lahore University of Management Sciences, Syed Babar Ali School of Science and Engineering, PAKISTAN
                Author notes

                Competing Interests: The J. Michael Bishop Institute of Cancer Research and Chengdu Anticancer Bioscience LTD has filed a PCT application (PCT/CN2021/82684) that covers the compound LXY18 under this study. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

                Author information
                https://orcid.org/0000-0002-4268-3394
                Article
                PONE-D-23-20900
                10.1371/journal.pone.0293283
                10615259
                37903144
                b72e4022-2334-4386-85d9-4f2bab5c9fe1
                © 2023 Kalashova et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 5 July 2023
                : 10 October 2023
                Page count
                Figures: 7, Tables: 0, Pages: 19
                Funding
                Funded by: The J. Michael Bishop Institute of Cancer Research receives funding through an endowment from Anticancer Bioscience, a company actively engaged in the commercial development of cancer therapeutics
                Award ID: 2016
                Award Recipient :
                The J. Michael Bishop Institute of Cancer Research receives funding through an endowment from Anticancer Bioscience in 2016, a company actively engaged in the commercial development of cancer therapeutics. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cell Death
                Apoptosis
                Medicine and Health Sciences
                Oncology
                Cancers and Neoplasms
                Medicine and Health Sciences
                Oncology
                Cancer Treatment
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cell Cycle and Cell Division
                Mitosis
                Biology and Life Sciences
                Cell Biology
                Chromosome Biology
                Mitosis
                Biology and Life Sciences
                Cell Biology
                Molecular Motors
                Motor Proteins
                Biology and Life Sciences
                Biochemistry
                Proteins
                Motor Proteins
                Medicine and Health Sciences
                Oncology
                Cancers and Neoplasms
                Lung and Intrathoracic Tumors
                Research and analysis methods
                Biological cultures
                Cell lines
                DU145 cells
                Medicine and Health Sciences
                Oncology
                Cancer Treatment
                Cytokine Therapy
                Custom metadata
                All relevant data are within the manuscript and its Supporting Information files.

                Uncategorized
                Uncategorized

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