eprintid: 10153569 rev_number: 15 eprint_status: archive userid: 699 dir: disk0/10/15/35/69 datestamp: 2022-08-11 11:17:26 lastmod: 2022-10-14 16:25:40 status_changed: 2022-08-11 11:17:26 type: article metadata_visibility: show sword_depositor: 699 creators_name: Dale, Katie L creators_name: Armond, Jonathan W creators_name: Hynds, Robert E creators_name: Vladimirou, Elina title: Modest increase of KIF11 exposes fragilities in the mitotic spindle causing chromosomal instability ispublished: pub divisions: C10 divisions: G99 divisions: B02 divisions: UCL divisions: D19 keywords: CRISPR activation, Chromosomal instability, Confocal imaging, Deep learning, KIF11, Mitosis note: © 2022. Published by The Company of Biologists Ltd This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. abstract: Chromosomal instability (CIN), the process of increased chromosomal alterations, compromises genomic integrity and has profound consequences on human health. Yet, our understanding of the molecular and mechanistic basis of CIN initiation remains limited. We developed a high-throughput, single-cell image-based pipeline employing deep learning and spot counting models to detect CIN by automatically counting chromosomes and micronuclei. To identify CIN-initiating conditions, we used CRISPR activation in human diploid cells to upregulate, at physiologically-relevant levels, 14 genes that are functionally important in cancer. We found that upregulation of CCND1, FOXA1, and NEK2 resulted in pronounced changes in chromosome counts and KIF11 upregulation resulted in micronuclei formation. We identified KIF11-dependent fragilities within the mitotic spindle; increased KIF11 causes centrosome fragmentation, higher microtubule stability, lagging chromosomes or mitotic catastrophe. Our findings demonstrate that even modest average single gene expression changes in a karyotypically stable background are sufficient for initiating CIN by exposing fragilities of the mitotic spindle which can lead to a genomically-diverse cell population. date: 2022-09 date_type: published publisher: The Company of Biologists official_url: https://doi.org/10.1242/jcs.260031 oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 1969394 doi: 10.1242/jcs.260031 medium: Print-Electronic pii: 276220 lyricists_name: Hynds, Robert lyricists_id: HYNDS02 actors_name: Flynn, Bernadette actors_id: BFFLY94 actors_role: owner funding_acknowledgements: C7893/A24956 [Cancer Research UK]; WT209199/Z/17/Z [Wellcome Trust]; [NIHR Great Ormond Street Hospital Biomedical Research Centre] full_text_status: public publication: Journal of Cell Science volume: 135 number: 17 article_number: jcs.260031 event_location: England citation: Dale, Katie L; Armond, Jonathan W; Hynds, Robert E; Vladimirou, Elina; (2022) Modest increase of KIF11 exposes fragilities in the mitotic spindle causing chromosomal instability. Journal of Cell Science , 135 (17) , Article jcs.260031. 10.1242/jcs.260031 <https://doi.org/10.1242/jcs.260031>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10153569/1/DownloadCombinedArticleAndSupplmentPdf.pdf