Inhibitors of Rho kinases (ROCK) induce multiple mitotic defects and synthetic lethality in BRCA2-deficient cells

 The trapping of Poly-ADP-ribose polymerase (PARP) on DNA caused by PARP inhibitors (PARPi) triggers acute DNA replication stress and synthetic lethality (SL) in BRCA2-deficient cells. Hence, DNA damage is accepted as a prerequisite for SL in BRCA2-deficient cells. In contrast, here we show that inhibiting ROCK in BRCA2-deficient cells triggers SL independently from acute replication stress. Such SL is preceded by polyploidy and binucleation resulting from cytokinesis failure. Such initial mitotic abnormalities are followed by other M-phase defects, including anaphase bridges and abnormal mitotic figures associated with multipolar spindles, supernumerary centrosomes, and multinucleation. SL was also triggered by inhibiting Citron Rho-interacting kinase, another enzyme that, similarly to ROCK, regulates cytokinesis. Together, these observations demonstrate that cytokinesis failure triggers mitotic abnormalities and SL in BRCA2-deficient cells. Furthermore, the prevention of mitotic ..., Flow Cytometer Data (FCS) was obtained by using an acoustic flow cytometer and autosampler (Attune NxT, Invitrogen). FCS data files were obtained by acquiring equal proportions of each well from a 96-well plate. Attune Software was used for gating. The same gating strategy has been used for all data files.
Automated analysis of Flow cytometry data
We developed a script using R to automatically import and analyze all the data from the screenings. C:\Users\54266\AppData\Local\Temp\{7F4D05D3-C49B-4D18-8DF9-5FE167F87F54}
 
, Attune Software or FlowJo and R.
, # Data from: Inhibitors of Rho kinases (ROCK) induce multiple mitotic defects and synthetic lethality in BRCA2-deficient cells

General README (ScreeningROCK.zip and All_excels_files.zip files)

GENERAL INFORMATION

Description of the data contained in each file:

In the database, you will find a zip (All_excels_files) containing the following files:

1\) All_excels_files.xls, containing all the raw data use to generate all plots presented in all figure panels from Martino et al., 2023. Each sheet contains the raw data of a single plot from a single figure as made explicit in the title of each sheet.

2\) The same data from All_excels_files.xls is contained in a folder (named: All_excels_files_csv) containing the exact same material from the excel files but saved as raw CSV data. Each CSV data file was generated from a single sheet from the excel files, using the command: SAVE AS “comma-separated values (CSV) files”.

3\) The data corresponding to the screening used to generate figure 1..., ,