Figure 2 from Concurrent RB1 Loss and <i>BRCA</i> Deficiency Predicts Enhanced Immunologic Response and Long-term Survival in Tubo-ovarian High-grade Serous Carcinoma
Authors: Saner Flurina A.M. , Takahashi Kazuaki , Budden Timothy , Pandey Ahwan , Ariyaratne Dinuka , Zwimpfer Tibor A. , Meagher Nicola S. , Fereday Sian , Twomey Laura , Pishas Kathleen I. , Hoang Therese , Bolithon Adelyn , Traficante Nadia , Alsop Kathryn , Christie Elizabeth L. , Kang Eun-Young , Nelson Gregg S. , Ghatage Prafull , Lee Cheng-Han , Riggan Marjorie J. , Alsop Jennifer , Beckmann Matthias W. , Boros Jessica , Brand Alison H. , Brooks-Wilson Angela , Carney Michael E. , Coulson Penny , Courtney-Brooks Madeleine , Cushing-Haugen Kara L. , Cybulski Cezary , El-Bahrawy Mona A. , Elishaev Esther , Erber Ramona , Gayther Simon A. , Gentry-Maharaj Aleksandra , Gilks C. Blake , Harnett Paul R. , Harris Holly R. , Hartmann Arndt , Hein Alexander , Hendley Joy , Hernandez Brenda Y. , Jakubowska Anna , Jimenez-Linan Mercedes , Jones Michael E. , Kaufmann Scott H. , Kennedy Catherine J. , Kluz Tomasz , Koziak Jennifer M. , Kristjansdottir Björg , Le Nhu D. , Lener Marcin , Lester Jenny , Lubiński Jan , Mateoiu Constantina , Orsulic Sandra , Ruebner Matthias , Schoemaker Minouk J. , Shah Mitul , Sharma Raghwa , Sherman Mark E. , Shvetsov Yurii B. , Soong T. Rinda , Steed Helen , Sukumvanich Paniti , Talhouk Aline , Taylor Sarah E. , Vierkant Robert A. , Wang Chen , Widschwendter Martin , Wilkens Lynne R. , Winham Stacey J. , Anglesio Michael S. , Berchuck Andrew , Brenton James D. , Campbell Ian , Cook Linda S. , Doherty Jennifer A. , Fasching Peter A. , Fortner Renée T. , Goodman Marc T. , Gronwald Jacek , Huntsman David G. , Karlan Beth Y. , Kelemen Linda E. , Menon Usha , Modugno Francesmary , Pharoah Paul D.P. , Schildkraut Joellen M. , Sundfeldt Karin , Swerdlow Anthony J. , Goode Ellen L. , DeFazio Anna , Köbel Martin , Ramus Susan J. , Bowtell David D.L. , Garsed Dale W. 
Published: August 2024

<p>Sensitivity to therapeutic agents in <i>BRCA1</i>-altered cell lines with <i>RB1</i> knockout. <b>A,</b><i>RB1</i> was knocked out using CRISPR/Cas9 in three patient-derived Australian Ovarian Cancer Study (AOCS) HGSC cell lines with either wild-type or altered <i>BRCA1</i> (<i>BRCA1</i> var) background. Representative Western Blots show protein levels of RB1 and phosphorylated RB1 (pRB1) compared with GAPDH loading control in single-cell cloned, homozygous <i>RB1</i> wild-type (WT) and knockout (KO) colonies in comparison with heterogeneous populations with a scramble single guide RNA (sgRNA). Independent blots were used for RB1 and pRB1. <b>B,</b> Cell viability was compared between <i>RB1</i> WT and KO clones following treatment with cisplatin (72 h), paclitaxel (72 h), or olaparib (120 h). Nonlinear regression drug curves are shown; <i>P</i> values are shown in Supplementary Table S18 (<i>n</i> = 3). Error bars indicate ± SEM; for some values, error bars are shorter than the symbols and thus are not visible. <b>C,</b> Proportion of surviving colonies following 16 days of treatment with cisplatin, paclitaxel, or a combination of both (Cis/Pac; with half of the IC50 determined per drug and cell line respectively) relative to DMF vehicle control (<i>n</i> = 3 replicates). Data are presented as mean ± SEM. Mean values were compared by Student’s <i>t</i> test (ns, not significant; *, <i>P</i> < 0.05; **, <i>P</i> < 0.01). Representative scans of the fixed cell colonies stained with crystal violet are shown for each condition.</p>